hoorieh soleimanjahi

In recent years, the collection of data from the monitoring of clinical trials in the form of electronic case report forms (eCRF) has been increasingly used, because avoiding mistakes, reducing the duration of clinical studies, and preventing increasing data collection costs can affect the success of the study. Accordingly, this research aimed to present the conceptual model and requirements of the proposed eCRF, which is based on NoSQL database to achieve complications caused by the COVID-19 vaccine.

In this study, using the descriptive-applicative method, the conceptual model of the eCRF system was first designed to collect the minimum set of data on the complications of the COVID-19 vaccine. Then, the functional and non-functional requirements of the proposed system were identified using the methods of observation, thinking and reflection, and questionnaire.

This research has two main parts: the proposed conceptual model and the functional and non-functional requirements of the eCRF system. A conceptual model was proposed based on the components, modules, and objectives of the eCRF core, which can be used by researchers and clinical trial specialists. Requirements including the ability to register, insert, delete, and edit the case report form, were also calculated for four groups of patients, health care workers, disease experts, and the executive manager.

The proposed system provides researchers and experts of clinical trials with classified and valid data regarding the registration of complications caused by the COVID-19 vaccine in an integrated manner. Therefore, some eCRF stakeholders such as patients, healthcare workers, disease experts, and executive managers and researchers can achieve a vaccine with acceptable safety and effectiveness by analyzing, interpreting, and sharing the recorded information.

Gene transfection is a powerful tool for changing cell function, which is used for treating various diseases. Nevertheless, various studies show that despite the variety of gene transfer methods, there are significant limitations in the application of each method. The present study aimed to compare two methods of gene transfer using PEI and Lipofectamine compounds in cancer cell.

In this study, Caco-2 and HCT-116 were used as target cells, and HEK-293 cells were used as control cells. For this purpose, initially, gene transfer was performed using PEI and Lipofectamine, and then their results were compared with each other based on the expression of GFP protein.

The results of this study showed that although gene transfer was carried out in HEK-293 cells at a favorable rate, this process was very insignificant in colorectal cells. In addition, in the meantime, some differences were observed between the amounts of gene transfer in colorectal cells, so that the amount of transfection in Caco-2 cells was lower than that of the HCT-116 cells.

The degree of acceptance of foreign genes in cells depends on several factors, including the type and origin of the cell. Since cancer cells have undergone many genetic changes, their genetic manipulation is associated with many limitations. For this reason, it is recommended to use other methods, such as cell selection by antibiotics and preparation of stable cell lines to reach a high level of gene transfer in these cells.

Group A Rotavirus (RVA) is the most important causative agent of acute diarrheal disease in pediatrics 5 years and below. This study aimed to determine the distribution of circulating RVA in Mashhad, Iran to develop health improvement strategies and vaccine decision making.

A total of 106 fecal specimens were collected from children admitted to Akbar and Dr. Sheikh referral pediatric hospitals of Mashhad City during the December 2020 to March 2021 and December 2021 to March 2022. All specimens were tested for specific bacterial, parasitic, and amoebic infections. Negative samples were analyzed for RVA infections using the RT-PCR method.

RVA was detected in 31.3% of the specimens, indicating no statistical significance in gender distribution or between fall and winter positivity rates. The number of RVA–positive specimens increased following age increasing in the range of 1 to 60 months.

Today, acute diarrheal disease (ADD) is still caused mostly by Rotavirus infections in pediatrics in Mashhad. Comprehensive studies are needed to determine the genetic diversity of circulating Rotavirus strains in this era.

HPV infections cause a wide spectrum of pathological changes in lower anogenital epithe- lium. The aim of this study was to investigate the HPV DNA status and histological findings in cervical biopsy specimens diagnosed as flat condyloma.

This study included 20 cervical biopsy specimens diagnosed as flat condyloma. The histopatholog- ical criteria and presence of HPV DNA were evaluated. HPV genotyping was determined in HPV-positive specimens using BioEdit software and the results were analyzed in SPSS software.

HPV DNA was not found in 30% of specimens and relative frequency of HPV genotypes was: 15% HPV6, 15% HPV11, 5% HPV16, 5% HPV18, 5% HPV53, 5% HPV68, 5% HPV84, 10% HPV45. Relative frequency of histopathologi- cal criteria was as below: 100% of specimens had koilocytosis, 100% acanthosis, 15% nuclear immaturity, 100% atypia, 15% mitotic activity, 50% dyskeratosis, 35% parakeratosis and 10% hyperkeratosis.

There were significant differences between HPV positivity and two pathologic criteria; multinucleation and hyperkeratosis (P Value: 0.02). Nuclear immaturity was significantly more prevalent in high risk HPV-positive specimens (P Value: 0.03).

SARS-COV-2 infection is not always correlated with protection. Antibody seroprevalence in unvaccinated individuals, which is usually measured by N-specific antibodies, is not necessarily correlated with protection, while antibodies against S protein show a better correlation with protection due to its neutralizing epitopes. In this study, we tried to improve our conception of the hidden perspective of SARS-COV-2 in epidemiological reports and investigate anti-S antibody prevalence among anti-N antibody-positive asymptomatic and mildly symptomatic patients.

Blood samples were collected from asymptomatic or mildly symptomatic volunteer participants and symptomatic hospitalized patients with negative PCR results from May 30 to June 17, 2020. Detection of SARS-COV-2 antibodies was done using an ELISA kit targeting N or S protein.

Totally, 716 samples from volunteer participants and 81 samples from symptomatic hospitalized patients with negative PCR results were evaluated. The test performance-adjusted seroprevalence (95% CI) of SARS-COV-2 antibody was 17.3% (8.8-25.8%) for anti-N IgG in volunteers and 25.5% (12.8-39.7%) for anti-N and anti-S IgM in hospitalized patients. Among anti-N IgG positive infected individuals, 49.2% (21.4 and 78.8%) were anti-S antibody positive.

The results showed that SARS-COV-2 infection sometimes occurs in individuals without symptoms or with mild symptoms, but in more than half of them, the produced antibody is not protective. The findings of hospitalized patients showed that the combination of IgM assay with real-time PCR improved the disease diagnosis by more than 25% in cases with negative molecular test results.

Exercise activities can act as an endogenous adjuvant, enhancing the efficiency of the host immune response after vaccination. However, the effect of repeated bouts of eccentric endurance exercise on cellular and humoral immune-related responses to the vaccine is unclear. The present study was performed to investigate the activation of type 1 and 2 (Th) helper lymphocytes following three bouts of endurance eccentric exercise as an adjuvant for DNA vaccination. Thirty-six female Balb-c mice (6-8 weeks, body mass mean=18g) were randomly divided into six groups: control, exercise, vaccine, PcDNA, exercise-vaccine and exercise-PcDNA (6 mice in each group). Different groups underwent DNA vaccine and PcDNA injection immediately after the last exercise, all stages of exercise and injections were repeated after 12 days according to the vaccine protocol, and 12 days after the second vaccine injection, spleen samples were collected. On-way variance analysis was used for statistical analysis and P<0.05 considered for significance. According to the results of the study, the ratio of Th1/Th2 after antigen stimulation was significantly different between the control group with vaccine and exercise-PcDNA (P <0.05). Also, the exercise-vaccine group was significantly different from all research groups (P <0.05). The results of the present study confirm the increase in the Th1/Th2 cell ratio following repeated bouts of eccentric endurance exercise. Stimulation of muscle damage through exercise before DNA vaccination may have been effective in invoking innate immune system cells and effectively stimulating acquired immunity.

Various infectious and non-infectious factors can cause encephalitis in the central nervous system (CNS), the most important of which are viruses. Herpes viruses are one of the most important causes of encephalitis worldwide. PCR detected the virus on the cerebrospinal fluid (CSF) sample. The aim of this study was to set up an in-house PCR to identify herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) and determine the prevalence of these viruses in suspected children of encephalitis.

This cross-sectional study was conducted on 160 suspected children with encephalitis cases referred to Dr. Kermanshahi Children Hospital, Kermanshah, Iran, from April to March 2021. CSF samples were extracted using a viral extraction kit, and a PCR was performed. The level of glucose and total protein of the samples was measured.

The total prevalence of HSV was 16.25%. 17 samples were positive for HSV-1 (10.6%), and 9 samples for HSV-2 (5.6%). There was a significant correlation between glucose, total protein, and HSV PCR positive, but no significant correlation between age and HSV PCR positive results.

Rapid diagnosis of a virus may reduce the hospitalization rate and the use of unnecessary therapies and crease mortality, morbidity, and disability in children. Results in this study show that the distribution of HSV types in children with encephalitis predominantly was type 1 compared with type 2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of coronavirus disease 2019 (COVID-19) as a pandemic infectious disease which has led to thousands of deaths around the world. The Coronaviridae family is the second cause of the common cold that targets human respiratory tracts. Specific diagnostic laboratory tests in addition to clinical investigations would be helpful in confirming COVID-19 in the early stages for controlling the disease. Upon SARS-CoV-2 infection, antibody responses are produced during the early phase of illness (>7 days), meanwhile, viral nucleic acid real-time reverse-transcription polymerase chain reaction (rRT-PCR) test is applied as the confirmatory assay in the first 5-6 days after the onset of illness. Due to the rise of antibodies, the viral nucleic acid represents a gradual decline. These laboratory tests may be considered valuable for monitoring the patient’s status to prevent the spreading of infections and keep him/her in quarantine. The results of molecular and serological assays revealed that whether the person is recovered and protected against disease. Furthermore, regarding the rise of antibody titer and undetectable viral RNA, it may be possible to make a decision about when the recovered people could back to work and social life.

Oncolytic reoviruses can infect and kill malignant cells while sparing their normal counterparts. Reoviral infection can induce or activate autophagy, even though metformin can induce autophagy. Identifying and regulating the cellular pathways important for reovirus replication and oncolysis can improve targeted-biological therapies for cancer. Here, the autophagic process was triggered via metformin, and we investigated the effect of autophagy activation on oncolytic reovirus replication in mesenchymal stem cells as primary cells and L929 cell lines.

Adipose derived mesenchymal stem cells (AD-MSCs) and L929 cells were treated with metformin and reovirus type-3 strain Dearing (T3D). Twenty-four hours after infection, the viability of AD-MSCs and L929 cells were examined by MTT assay. Also, the effect of metformin-induced autophagy in the reovirus replication in these cells was determined by real-time polymerasechain-reaction.

Our results show that treatment with metformin and reovirus reduced the viability of the cells compared to treatment with metformin or reovirus alone in both cells. Also, coadministration of metformin and reovirus significantly decreased the relative expression level of the Beclin-1 gene compared to treatment with metformin in both cells. However, the expression level of the reovirus L3 gene after treatment with metformin and reovirus in L929 cells increased significantly compared to AD-MSCs.

Our data suggest that metformin-induced autophagy enhances reoviral replication in AD-MSCs and L929 cells. These findings represent the role of autophagy induction in facilitating reovirus replication and contribute to a better understanding of reovirus-host interactions.

One of the major concerns in the world is the Coronavirus Disease 2019 (COVID-19), an infectious disease caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2). Although the knowledge is limited about the risk factors for the disease, morbidity and mortality may increase in the patients with some underlying conditions such as cardiovascular disease, hypertension, diabetes, cancer, etc. These diseases can weaken the immune system and affect the body's ability to respond to infectious agents. Therefore, these patients are more at the risk for COVID-19 and also the underlying condition may worsen the severity of COVID-19 infection. On the other hand, SARS-CoV-2 via multiple pathophysiological mechanisms can lead to the progression of the underlying diseases and resulting in a poor outcome. The coronavirus binds to angiotensin-converting enzyme 2 (ACE2) which is expressed on the cells of many organs and it can directly affect tissues. Apoptosis of the cells may occur in patients with acute respiratory disease syndrome (ARDS) due to hypoxia in COVID-19. Moreover, SARS-CoV-2 leads to an imbalanced the immune inflammatory response in some patients which may cause indirectly organ injury. In this review we described the prevalence of COVID-19 in various underlying diseases and the impact of the SARS-CoV-2 in the outcome of these diseases. However, further studies are needed to investigate the prevalence of this new virus in patients with underlying diseases and its effects on the progression of illness.

Human papillomavirus (HPV) is a primary contributing agent of cervical cancer. Eradication of HPV-related infections requires therapeutic strategies. We used Brucella abortus RB51 rough lipopolysaccharide (R-LPS) as an adjuvant along with two HPV16 therapeutic DNA vaccines, pcDNA3-E7 and pcDNA3-L1, for improving DNA vaccine efficacy. For evaluation of the B. abortus LPS adjuvant efficacy in combination with DNA vaccines to induce cellular immune responses, C57BL/6 mice were immunized with the DNA vaccines, with or without R-LPS adjuvant. IFN-γ and IL-4 cytokines assay was carried out for assessment of cellular and humoral immune responses.   Findings indicated that vaccination with pcDNA3-E7 or pcDNA3-L1 alone could induce strong cellular immune responses, but stronger antigen-specific T-cell immune responses were shown by co-administration of HPV16 E7 and HPV16 L1 DNA vaccines along with R-LPS adjuvant. Overall, B. abortus R-LPS through enhancement of T-cell immune responses can be considered an efficient vaccine adjuvant in future studies and trials.

Coronaviridae family cause respiratory diseases ranging from common cold to severe Respiratory diseases such as SARS, MERS and new emerging coronavirus disease COVID-19.  Also the family including four other human coronaviruses (HCoV-229E, HCoV-HKU1, HCoV-NL63, and HCoV-OC43) with flu likes symptoms. Coronaviruses cannot be distinguished clinically from other respiratory infectious agents. Based on the health importance and widespread distribution of respiratory infections, the current study was designed for diagnosis of Pancoronaviruses.  

Nasopharyngeal swabs from 200 patient suspected viral upper respiratory tract infection analyzed using optimized RT-PCR assay.  The constructive specific degenerate primers were used for amplification of rep1b ORF from coronaviruses genome. The 354bp DNA fragment related to 229E coronavirus polymerase gene was amplified from Amplirun Total Respiratory Viral Panel Control (Vircell) template by RT-PCR. Amplified product ligated into T-easy vector (Promega). Plasmid then transformed to Top 10 Fchr('39') strain by chemical method and Positive colonies were selected using colony PCR with gene specific primers. Diagnostic restriction enzyme digestion was done with EcoRI restriction enzyme. Vector was linearized by SacI restriction enzyme and In-vitro transcription was performed using TranscriptAid T7 High Yield Transcription Kit. DNA was removed with DNase I treatment. Then the detection limit of the specific Rep1b primers was determined by Two- Step RT-PCR with synthetic RNA concentration gradient. All of samples were negative for Pancoronavirus.

All of these samples were negative for Pancoronavirus.

Larger sample sizes and proper sampling procedure may improve the chances of viral RNA detection.

A growing area of research is focused on cancer therapy, and new therapeutic approaches are welcomed. Mesenchymal stem cell (MSC)-based gene therapy is a promising strategy in oncology. Intrinsic tropism and migration to tumor microenvironment with off lights are attractive features of this type of cell carrier. In this way, suicide genes have also found a good platform for better performance and have shown a stronger anti-tumor mechanism by riding on mesenchymal cells. In this study, we investigated the anti-tumor activity of intratumoral injected MSCs transduced with a lentivector expressing the HSV/TK in a mouse cervical cancer model. Following the injection of MSCs transduced with lentivector carrying TK, MSCs alone or PBS into the mice tumor, ganciclovir was administered intraperitoneally during 14 days, and tumor size, survival time, natural killer (NK) cells and cytotoxic T lymphocyte (CTL) activities were assessed. We demonstrated that combination of suicide therapy and cell therapy leading m,to successful tumor inhibition. Significant reduction in tumor size was detected in test group in comparison with controls. Also, potent antitumor NK and CTL activity was seen in treatment group in comparison with controls. Our data demonstrated that the mesenchymal cells expressing TK had inhibitory effect on cervical cancer model.

Enzymatic digestion is an essential stage for culturing Mesenchymal Stem Cells (MSCs) and their therapeutic application. Several factors such as being cost-benefit, efficiency, safety, yield and amount of produced cells are determinant for choosing the appropriate enzyme. Collagenase is a conventional enzyme commonly used for enzymatic digestion. However, other enzymes like trypsin and even combination of these enzymes can be used as an alternative strategy in different situations.

Abdominal subcutaneous adipose tissue was obtained from male BALB/c mice and digested under three different enzymatic processes: collagenase and collagenase/trypsin and trypsin. Cell culture process was performed under standard condition and MSCs at 3rd passage were used for further characterization by flow cytometry.

In this study, two different enzymatic methods for digestion of adipose-derived MSCs (ADSCs) of BALB/c mice were investigated. The morphology of cells was pretty different and was more homogenous in collagenase group. Also the yield of cells was varied among groups. Furthermore, the obtained data from flow cytometry revealed that ADSCs were positive for CD90 (70%), CD29 (98%), CD105 (52%) and negative for CD45 (<2%).

Application of different enzymes depends on various conditions. Altogether, these data indicate that although use of trypsin in isolation protocol is cost-benefit, it can be used as an alternative method whenever limited number of cells will be needed. However, collagenase as a well-known and conventional method can be used for isolation of larger quantity of cells with several applications.

SARS-COV-2, the latest member of Coronaviridea family as the cause of the recent global epidemic, has inflicted sever damages in different fields on most governments. Appropriate strategies such as identifying infected individuals and isolating them, trying to produce an effective vaccine, finding efficient treatment and making correct decisions in the social, economic and health fields are the current priorities of the world. Serological tests provide useful information in all of the above areas. In this article we will discuss the importance of performing SARS-COV-2 serological tests in diagnosis, vaccination, treatment and proper planning in the society.

Currently, application of oncolytic-virus in cancer treatment of clinical trials are growing. Oncolytic-reovirus is an attractive anti-cancer therapeutic agent for clinical testing. Many studies used mesenchymal stem cells (MSCs) as a carrier cell to enhance the delivery and quality of treatment with oncolytic-virotherapy. But, biosynthetic capacity and behavior of cells in response to viral infections are different. The infecting process of reoviruses takes from two-hours to one-week, depends on host cell and the duration of different stages of virus replication cycle. The latter includes the binding of virus particle, entry, uncoating, assembly and release of progeny-viruses. We evaluated the timing and infection cycle of reovirus type-3 strain Dearing (T3D), using one-step replication experiment by molecular and conventional methods in MSCs and L929 cell as control.

In this experimental study, L929 and adipose-derived MSCs were infected with different multiplicities of infection (MOI) of reovirus T3D. At different time points, the quantity of progeny viruses has been measured using virus titration assay and quantitative real-time polymerase chain reaction (qRT-PCR) to investigate the ability of these cells to support the reovirus replication. One-step growth cycle were examined by 50% cell culture infectious dose (CCID50) and qRT-PCR.

The growth curve of reovirus in cells shows that MOI: 1 might be optimal for virus production compared to higher and lower MOIs. The maximum quantity of virus production using MOI: 1 was achieved at 48-hours post-infection. The infectious virus titer became stationary at 72-hours post-infection and then gradually decreased. The virus cytopathic effect was obvious in MSCs and this cells were susceptible to reovirus infection and support the virus replication.

Our data highlights the timing schedule for reovirus replication, kinetics models and burst size. Further investigation is recommended to better understanding of the challenges and opportunities, for using MSCs loaded with reovirus in cancer-therapy.

Cytomegalovirus End-Organ Disease (CMV-EOD) is a seriously debilitating illness in patients with advanced HIV-1 infection, typically occurring with CD4+ cell counts of < 100 cells/mm3.

This study aimed to evaluate the prevalence of CMV-EOD in adult patients with advanced HIV-1 infection (CD4+ count < 100 cells/mm3).

Using a convenience sampling method, a cross-sectional study was conducted on 82 patients with advanced HIV-1 infection in Iran between April 2016 and April 2018. We collected baseline characteristics (age, sex, route of HIV-1 transmission, Hepatitis C Virus (HCV) infection, Hepatitis B Virus (HBV) infection, CMV IgG, and treatment status for HIV-1 infection) and CD4 counts. The entire patients underwent clinical examinations for the diagnosis of CMV-EOD by experienced clinicians. Statistical analysis was used to measure the differences between categorical variables and the outcome of CMV-EOD diagnosis.

Fourteen (17.07%) out of 82 HIV-1-infected patients were diagnosed with opportunistic infection due to CMV. Among 14 patients with CMV-EOD, retinitis occurred in the majority of patients (64.28%), followed by colitis (21.42%) and encephalitis (14.28%). No significant correlation was found between the outcome of CMV-EOD and HBV infection (P = 1.00), HCV infection (P = 0.55), and treatment status for HIV-1 infection (P = 0.53). We detected CMV-EOD more frequently among injecting drug users and patients with positive CMV-IgG (P = 0.12 and P = 0.41, respectively). The ophthalmic examination had clinical usefulness for HIV-1 positive patients with CD4 counts of < 100/mm3.

It is assumed that the CD4+ cell count is not the sole predictor of the risk of developing CMV-EOD. Further large-scale studies are required for a better understanding of risk factors involved in the occurrence of CMV-EOD in HIV-1 positive patients.

Autophagy has been introduced as a protective mechanisms in cell in recent years. Regarding to the changes induced in autophagy factors following an acute physical exercise. The purpose of this study was to investigate the changes in autophagy factors in time courses of acute exhaustive endurance exercise in Tibialis Anterior skeletal muscle of BALB/c mice.

Regarding the purpose of the study, 12 BALB/c mice were divided into two groups of exercise and control. Subjects were anatomized immediately and 12 hours after the end of the exercise session. The Real Time-PCR method was used to determine the expression of autophagy factors.

Independent t-test showed significant increases in autophagy activation immediately after exercise and significant decreases 12 hours following acute exhaustive endurance exercise (P≤0.05).

  Our results proposed that significant increases induced in autophagy factors immediately after acute exhaustive exercise. Possible correlations between autophagy activation and changes in oxidative stress, some immunological factors and metabolic responses could be some of mechanisms of autophagy activation following acute exercise.

During the 15 years since the discovery of type III human interferons [IFN-λ1(IL-29), IFN-λ2(IL-28A), and IFN-λ3(IL-28B)], numerous biological properties such as anticancer, antiviral, and immunomodulatory activities of this new IFN family have been investigated. Several studies have shown that the encapsulation of pcDNA with PLGA nanoparticles (NPs) protects them against DNase enzyme action and increases the efficiency of gene delivery to the cells. The purpose of this study was to encapsulate pcDNA encoding IFN-λ1 (pIFN-λ1) with a simple and cost-effective method using PLGA NPs. The pIFN-λ1-loaded PLGA NPs were produced by a double-emulsion-solvent evaporation method and characterized in terms of size, size distribution, and zeta potential by DLS and morphologically by SEM and TEM. The bioactivity of NPs was also examined by fluorescent microscopy. The results showed that IFN-λ1 expressed by HEK293T cells could protect HepC-2 cells from the cytopathic effects of EMCV. The NPs were spherical in shape with a mean diameter of 380 ± 3 nm, a zeta potential of −3.3 ± 7.6 mV, an encapsulation efficiency of 75 ± 5%, and a loading capacity of 0.83 ± 0.06. The NPs were also bioactive and easily engulfed by RAW264.7 cells. The pIFN-λ1 could be sustainably released from NPs. Due to the facility and affordability of encapsulation of pIFN-λ1 in the PLGA NPs proposed in this study and the advantages of encapsulation by PLGA, it appeared rational to use pIFN-λ1-loaded NPs instead of naked pIFN-λ1 to determine other unexplained activities of this new cytokine or to use it as an alternative or adjunct to current IFN-α therapy.

Cervical cancer is an important cause of death in women worldwide (1, 2). Cancer is a disease that may be caused by many factors that affect gene activity through genetic and epigenetic changes like DNA methylation. DNA promoter methylation contributes to the chromatin conformation that may be repressing transcription of the human papilloma virus type16 (HPV-16), which is prevalent in the etiology of cervical carcinoma. In the present study, we aimed to investigate DNA methylation target sites in promoter region of both high-risk and low risk HPVs. Methylation pattern of E6 promoter in low-risk HPV (type 11) and high-risk HPV (type 16 and 18) was examined by Bisulfite Sequencing PCR (BSP) method. Based on the results, methylation status of high-risk and low-risk HPV-E6 promoter is different. It was revealed that CpG dinucleotides were unmethylated in type 16 and 18 target sequences, whereas in HPV-E6 type 11 all of CpG dinucleotides were methylated except one of them. The result suggested that a significant correlation between methylation status and HPV-induced cervical carcinogenesis, and promoter of HPV-16 and 18 E6 has minimal methylation in comparison with low-risk HPV-11.

Oncolytic viruses (OVs) are a new approach in treatment of cancer. Antitumor efficacy of OVs were limited due to insufficient and non-specific viral delivery to tumor sites. To overcome this issue, mesenchymal stem cells (MSCs) were used for their ability to specifically homing into tumors. The main aim of this study was to use MSCs as carriers and investigate the effect of oncolytic reovirus infection in MSCs, induction of apoptosis, nitric oxide (NO) secretion and their effects for selectively killing tumor cells, to use in future. MSCs isolated from mice adipose tissue and confirmed. Then, the ability of the virus to infect MSCs and the effect of reovirus infection in induction of apoptosis and NO secretion in MSCs were evaluated. The results demonstrate that reovirus could replicate on MSCs. The finding indicated that the NO production significantly was higher at 72 h post infection with different MOI in comparison to the control cells. Also, reovirus induced high level of apoptosis in the MSCs at 48 h post infection compared with the control cells. Based on observed results, reovirus increased the secretion of iNOS (inducible nitric oxide) in the infected MSCs at 48 h post infection; therefore, high amounts of NO and reovirus replication were found to trigger apoptosis at 48 h post infection. Therefore, by optimizing the replication time of virus in the MSCs, specific viral delivery to tumor sites are available and causes cancer cells’ death.

Millions of people in developing countries lose their lives due to acute respiratory infections, such as Influenza A & B and Adeno viruses. Given the importance of rapid identification of the virus, in this study the researchers attempted to design a method that enables detection of influenza A, B, and adenoviruses, quickly and simultaneously. The Multiplex RT PCR method was the preferred method for the detection of influenza A, B, and adenoviruses in clinical specimens because it is rapid, sensitive, specific, and more cost-effective than alternative After collecting samples from patients with respiratory disease, virus genome was extracted, then Monoplex PCR was used on positive samples and Multiplex RT-PCR on clinical specimens. Finally, by comparing the bands of these samples, the type of virus in the clinical samples was determined. Performing Multiplex RT-PCR on 50 samples of respiratory tract led to following results; flu A: 12.5%, fluB: 50%, adeno: 27.5%, negative: 7.5%, and 2.5% contamination. Reverse transcription-multiplex Polymerase Chain Reaction (PCR) technique, a rapid diagnostic tool, has potential for high-throughput testing. This method has a significant advantage, which provides simultaneous amplification of numerous viruses in a single reaction. This study concentrates on multiplex molecular technologies and their clinical application for the detection and quantification of respiratory pathogens. The improvement in diagnostic testing for viral respiratory pathogens effects patient management, and leads to more cost-effective delivery of care. It limits unnecessary antibiotic use and improves clinical management by use of suitable treatment.

Vaccines based on virus-like particles are effective against Human Papilloma Virus (HPV) infection; however, they have not shown a therapeutic effect against HPV-associated diseases. New immunotherapy strategies based on immune responses against tumor antigens can positively affect the clearance of HPV-associated lesions.
To generate two therapeutic fusion DNA vaccines (optimizedE7/mouseHSP70 and wildE7/mouseHSP70) to induce antitumor specific responses in mice models.
Mice were immunized with recombinant DNA vaccines. The splenocytes of immunized mice were collected and lactate dehydrogenase and IFN-γ productions were measured after three injections in order to evaluate cytotoxic T lymphocytes (CTLs) activity. MTT assay was carried out for lymphocyte stimulation.
The fusion DNA vaccines, specifically uE7-HSP70, elicited varying levels of IFN-γ and CTLs responses compared to the control group (P It is concluded that our fusion DNA vaccines considerably enhanced specific cellular responses against HPV tumor model. In addition, optimized E7 showed a notable immunogenicity and inhibitory effect on the reduction of tumor size.

Human papillomavirus (HPV) oncoproteins, including E6 and E7 are constitutively expressed in cervical cancer cells. These proteins are ideal targets to be used for developing therapeutic vaccines against existing HPV-associated carcinomas. The aim of this study was to measure the proliferation response rate of splenic lymphocytes derived from E7-HPV16 encoding plasmid injection on the tumor mouse model of papillomavirus.
C57BL/6 mice were inoculated subcutaneous with 5× 10⁵ TC-1 cells in three times with two weeks intervals and then immunized with HPV-16 E7 DNA vaccine. The proliferation response of splenic cells was measured by MTT assay. IL12 cytokine was measured by ELISA assay and the mass of tumor was calculated with caliper for six weeks.
Following the application of DNA vaccines containing E7 therapeutic gene, the proliferative response of splenic cells was provoked significanltly higher than the stimulation in control group (P Our findings revealed that the application of DNA vaccine containing E7 gene in a tumor mouse model may induce anti-tumor cellular immune responses.

Cervical cancer is one of the main causes of women’s death in the world. Human papilloma virus (HPV) types 16 and 18 have been known as a cause of more than 2/3 of cervical cancers. New combination treatment strategies based on immune responses against viral antigens are likely to be beneficial. Considering the important role of angiogenesis in nutrition and oxygenation of tumor cells, the inhibition of this process can help tumor clearance.
In this study, co-administration of a plasmid encoding immune stimulatory epitopes of E6-E7-L1 genes of HPV with an anti-angiogenic peptide derived from Endostatin in tumor mice model was examined.
C57BL/6 mice were injected subcutaneously with TC-1 tumor cells and monitored for tumor progression. At exact time points, tumor size was determined. Then they were injected by pIRES-E6/E7/L1 as DNA vaccine, and anti angiogenic peptide. Relative tumor volume measurements and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was carried out in order to investigate therapeutic antitumor effects of vaccine and peptide.
Based on the results, the group of mice that received vaccine and vaccine-peptide had significant inhibition rate of tumor growth in comparison with control groups (P In general, it could be concluded that the co-administration of DNA vaccine due to the good immunogenicity and suitable inhibitory effect of peptide in reduction of tumor size, shows higher efficacy and can be considered as a new therapeutic strategy.