Management of organophosphorus poisoning and the role of magnesium sulfate: A scoping review of literature.

Organophosphorus agents are easily absorbed via respiratory, gastrointestinal, and dermal routes, and inhibit the acetylcholine transferase enzyme (AChE), which is responsible for the majority of toxicity caused by organophosphates in the body. A comprehensive search was conducted across three prominent databases, namely Google Scholar, PubMed, and Science Direct, to identify relevant articles published. The search focused on the keywords "MgSO4" or "magnesium sulfate" in conjunction with "organophosphate" or "organophosphate poisoning." Inhibition of AChE results in the accumulation of acetylcholine (ACh) in synapses and stimulation of cholinergic receptors. Considering that several studies have shown the use of magnesium sulfate (MgSO4) in inhibiting the release of ACh in the central and peripheral sympathetic and parasympathetic synapses, this study was conducted to review the role of MgSO4 in the treatment of OP. The intravenous administration of MgSO4 exhibits favorable tolerability and clinical efficacy in alleviating cardiac toxicity associated with OP exposure.

Epigallocatechin-3-gallate maintains Th1/Th2 response balance and mitigates type-1 autoimmune diabetes induced by streptozotocin through promoting the effect of bone-marrow-derived mesenchymal stem cells.

Stem-cell-based therapy is one of the most promising therapeutic strategies owing to its regenerative and immunomodulatory properties. Epigallocatechin-3-gallate (EGCG), a known antioxidant and anti-inflammatory agent, has beneficial effects on cellular protection. We aimed to elucidate the feasibility of using EGCG, along with bone marrow-derived mesenchymal stem cells (BM-MSCs), to improve pancreatic damage through their immune regulatory functions in an experimental model of type 1 diabetes mellitus (T1DM) induced by multiple injections of streptozotocin (STZ). BM-MSCs were isolated from C57BL/6 mice and characterized. The diabetic groups were treated intraperitoneally with PBS, MSCs, EGCG, and a combination of MSCs and EGCG. Real-time PCR assays showed that MSCs with EGCG modulated T-bet and GATA-3 expression and upregulated the mRNA levels of Foxp-3 more efficiently. Analyses of spleen-isolated lymphocytes revealed that combinational treatment pronouncedly increased regulatory cytokines and decreased pro-inflammatory cytokines and splenocyte proliferation. The histopathological assessment demonstrated that co-treatment significantly reduced insulitis and recovered pancreatic islet morphology. Furthermore, the combination of MSCs and EGCG is associated with downregulated blood glucose and enhanced insulin levels. Therefore, combined therapy with EGCG and MSCs holds clinical potential for treating T1DM through synergetic effects in maintaining the Th1/Th2 response balance and promoting the regeneration of damaged pancreatic tissues.

Walking Ability, Participation, and Quality of Life in Children with Spastic Diplegic Cerebral Palsy: A Path Analysis Study.

Objectives: This study aims to design a conceptual model for the effect of various factors on walking ability, participation, and quality of life in children with spastic diplegic cerebral palsy (SDCP) and test it based on field data using path analysis. Materials & Methods: This cross-sectional study was performed on 181 children with SDCP. The following were used to measure each of the variables: the Modified Ashworth Scale, the Micro Manual Muscle Tester, the Timed Up and Go Test (TUG), the Boyd and Graham test, the goniometer, weight and height, the Gross Motor Function Classification System, the Life Habits Questionnaire, and cerebral palsy Quality of Life Questionnaire for Children. The structural model was tested in Amos 17. Results: All paths of the proposed model were significant (P <0.05). Among evaluated variables, muscle strength (B = -0.466), balance (B = 0.326), and spasticity (B = 0.143) affected walking ability. Moreover, as an intermediate factor, walking ability affected the subjects' participation (B = -0.819) and quality of life (B = -0.183). Conclusion: Muscle strength, balance, and spasticity are the most influential factors in the walking ability of children with SDCP. Furthermore, walking ability and participation are two critical factors in promoting the quality of life of these children.

Immune dysregulation and pathogenic pathways mediated by common infections in rheumatoid arthritis.

Rheumatoid arthritis (RA) is one of the world's most prevalent inflammatory autoimmune diseases, affecting between 0.4 and 1.3% of the population. The susceptibility to RA appears to be influenced by a complex interaction between a favorable genetic background and the existence of a specific immune reaction against a wide range of environmental variables. Among the known environmental variables, infections are believed to have a significant role in promoting the formation of autoimmune disorders, which are frequently caused by specific microorganisms. Infections have been linked to RA in recent medical studies. In this study, we selected the most prevalent infections associated with RA from the literature and described the data confirming their pathogenic role in RA. Our investigation included Mycobacterium, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Proteus mirabilis, Epstein-Barr virus, parvovirus, and Prevotella copri.

Association between disease-modifying antirheumatic drugs and bone turnover biomarkers.

Rheumatoid arthritis (RA) has been linked to an increased risk of osteoporosis as well as fractures. Patients diagnosed with RA had a 25% increased risk of osteoporotic fracture, according to a recent population-based cohort study that compared them to people without RA. Several studies have found a correlation between osteoporosis and the presence of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1, and 6. These cytokines play a crucial part in the process of bone resorption by boosting osteoclast activation and encouraging osteoclast differentiation. Based on the correlation between RA, osteoporosis, and inflammation, it is possible that systemic immunosuppression with disease-modifying antirheumatic drugs (DMARDs) can help individuals with RA have a lower chance of developing osteoporosis and osteoporotic fractures. There is little information on how different DMARDs, biologic or non-biologic, affect RA patients' bone metabolism. In this study, we present an overview of the influence that targeted therapies, such as biologics, non-biologics, and small molecule inhibitors, have on bone homeostasis in RA patients.

Toxoplasmosis Frequency Rate in Rheumatoid Arthritis Patients in Northeastern Iran.

Background: Toxoplasmosis is a zoonotic disease caused by the parasite Toxo-plasma gondii, a cosmopolitan intracellular parasite. It can be a risk factor for auto-immune diseases, including rheumatoid arthritis (RA). This study was designed to investigate the possible association between serological history of T. gondii infection and defined clinical manifestation of RA in Northeast of Iran. Methods: Overall, serum samples were collected from 50 RA patients and 40 healthy controls, from Qaem Hospital in Mashhad City, northeastern Iran in 2018. Seroprevalence of T. gondii infection was determined by ELISA. Results: The prevalence of anti -T. gondii IgG in RA patients 48% (24.50) was significantly higher than the control group 10% (4.40) (P <0.001). Erythrocyte sedimentation rate (ESR), anti-cyclic citrullinated peptide (anti-CCP) and (rheumatoid factor) RF levels between the RA and control groups (P <0.01). Control group were matched with patients for age, gender and living area. Conclusion: Given that a high correlation has been demonstrated between positivity rate of anti-T. gondii IgG and RA in Northeastern Iran, further studies will be necessary to clarify the pathogenesis of T. gondii among these patients.

Association between Serum Progastrin Biomarker Level and Gastric Cancer.

BACKGROUND AND OBJECTIVE: gastric cancer is the fifth most prevalent cancer and the fourth cause of death because of cancer. In Iran, northern and northwestern regions are considered gastric cancer hot spots. Identifying serum biomarkers could be helpful in early diagnosis of patients with gastric adenocarcinoma (GAC). Increase in progastrin level has been reported in different cancers. Given the diagnostic value of this biomarker, this study aimed to determine the diagnostic role of progastrin serum biomarker in patients with gastric cancer. METHODOLOGY: In this case-control study, forty patients with gastric cancer who were diagnosed by endoscopy and pathologic findings and visited Mazandaran Comprehensive Cancer Center. The participants had received no treatment yet and entered this study. The participants in case group were compared with the control group including forty-two individuals with no history of gastrointestinal cancer in their first-degree relatives and visiting the lab for routine tests. Progastrin serum level was assessed using ELISA kit. The Kruskal-Wallis test and Mann Whitney test, both non-parametric) were used for statistical analysis and the relation between the variables was examined using Pearson's correlation coefficient at 95% confidence level in SPSS 16. FINDINGS: In this study, progastrin serum level was significantly higher in patients with gastric cancer compared with normal participants (P = 0.035). Progastrin serum level had no significant relation with tumor clinicopathologic parameters (p-value > 0.05). CONCLUSION: Increase in progastrin may be utilized as a predictive factor for gastric cancer.

Harmaline downregulates angiogenesis markers and suppresses the growth of 4T1 breast cancer cells in vivo and in vitro.

The anti-angiogenic effects of harmaline, an alkaloid with emerging anti-tumor properties, are under investigation. In the present study, the effects of different doses of harmaline, either alone or in combination with doxorubicin (DOX), were assessed in mice models of breast tumor. Breast tumors were created by the subcutaneous injection of 4T1 cells into Balb/c mice. The mice received either normal saline, harmaline alone (10, 20, or 30 mg/kg), or harmaline (20 mg/kg) + DOX (10 mg/kg). Immunohistochemistry, ELISA, and real-time PCR were conducted to measure target parameters. Harmaline significantly increased tumor cells' sensitivity to DOX as confirmed by a significantly reduced tumor volume in the harmaline + DOX group after 24 days (P < 0.05). Also, the levels of Ki-67 (P < 0.001), MMP-2 (P < 0.001), and VEGF (P < 0.001) significantly decreased while the level of E-cadherin increased (P < 0.001) in the tumor tissues of the mice treated with 20 or 30 mg/kg harmaline or harmaline (20 mg/kg) + DOX (10 mg/kg) compared to the control group. There was a significant reduction in the serum level of IL-4 in tumor-bearing mice treated with harmaline (P < 0.05), and IFN-γ serum level was significantly augmented in all experimental groups compared to the control group (P < 0.05). The genes encoding VEGF, VEGF receptor 2, CD105, and COX2 were significantly down-regulated (P < 0.05 for all) in harmaline-treated (either alone or in combination with DOX) mice. In conclusion, harmaline seems to have the potential to be used as an anticancer agent for treating breast cancer.

Bilateral Wrist Drop Due to Lead Poisoning in a Young Woman With Opium Addiction.

Opium addiction can cause symptoms in the central or peripheral neurological systems, as well as gastrointestinal disorders and anemia; in such situations, lead poisoning should be considered and chelation therapy should be started as soon as possible. In adults, lead poisoning is an unusual cause of abdominal pain. A common form of lead neuropathy includes weakness of the wrist and finger extensors. We describe a 24-year-old female who developed severe lead poisoning after 3 years of opium consumption, leading to gastrointestinal complaints and bilateral wrist drops.

Immune-related adverse events (irAEs) in ankylosing spondylitis (AS) patients treated with interleukin (IL)-17 inhibitors: a systematic review and meta-analysis.

BACKGROUND: Ankylosing spondylitis (AS) is a chronic inflammatory rheumatic disease characterized by immune system dysregulation and inflammation in the joints. Interleukin (IL)-17 inhibitors are new biological drugs used to treat AS. In this study, we aimed to assess the risk of immune system-related AEs due to targeting IL-17 or IL-17R. METHODS: The CENTRAL, PubMed, Scopus, Google Scholar, Clinical Trials Registry, and ICTRP were searched for randomized clinical trials (RCTs) and non-RCTs until February 2021. The risk of irAEs in patients treated with IL-17 inhibitors compared to the placebo or a drug-free control was evaluated. In studies that reported AEs of the IL-17 inhibitors at several different time points, we compared the number of cases/100 patient-year in which irAEs were reported. Subgroup analyses were also performed based on the dose and type of drugs. RESULTS: Thirteen studies of 1848 AS patients treated by IL-17 inhibitors (secukinumab, ixekizumab, bimekizumab, and netakimab) and 764 participants who received a placebo were included. The risk of some AEs related to immune function in patients under IL-17 inhibitors treatment was significantly higher than that of the placebo group, including infection and infestation (risk difference RD = 0.09, P = 0.02), nasopharyngitis (RD = 0.04, P < 0.001), opportunistic infections (RD = 0.01, P = 0.04), and neutropenia (RD = 0.04, P = 0.03). Besides, the results of the Cochran Q test showed that there were significant differences between the occurrence of some AEs over time, including infection and infestations (p < 0.001, RCTs), upper respiratory tract infections (p < 0.001, non-RCTs), urinary tract infections (p < 0.001, non-RCTs), and diarrhea (p < 0.01, RCTs). CONCLUSIONS: The most common immune system-related AEs in patients treated with IL-17 inhibitors are mucosal and opportunistic infections.

Occurrence of hypoglycemia in patients with benzodiazepines poisoning: A cross-sectional study.

BACKGROUND: Benzodiazepine (BZD) poisoning is a common medical condition often accompanied by respiratory arrest, aspiration pneumonitis, and may rarely result in death. Although it is not studied well, hypoglycemia is a real and life-threatening condition that may occur in BZD poisoning. Thus, the present study was shown to determine the prevalence of hypoglycemia among patients with BZD poisoning compared to toxicity with other drugs. PATIENTS AND METHODS: In this study, patients with drug-induced poisoning referred to Vasei hospital, Sabzevar, eastern Iran, were registered and their blood glucose levels at presentation were collected during 2019. RESULTS: Overall, 300 poisoned patients were evaluated (17% with BZD poisoning). The prevalence of hypoglycemia was significantly higher in patients with BZD poisoning (11.8% vs. 2.8%, p = 0.004), especially in males (P = 0.016), aged >30 years old (p = 0.006). However, the prevalence of hypoglycemia was higher in patients with GCS<10 (p = 0.005) and in patients referred later than 1 h to a medical center (p = 0.015). CONCLUSION: Results were show that hypoglycemia is a frequent complication of BZD poisoning and should be considered in the initial evaluation of patients referred to the emergency department with this medical condition.

Toxoplasma gondii infection in patients with rheumatoid arthritis and systemic lupus erythematous diseases: serological and molecular evidence.

This study aimed to determine Toxoplasma gondii infection in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients in Mazandaran Province, north of Iran. During April 2019 to February 2020, 305 patients with RA (N=272) and SLE (N=33) diseases are included in this cross-sectional study. The baseline data of patients were collected into a questionnaire. Also, blood sample was taken from each patient in tubes containing k2-EDTA as anticoagulant for serological and molecular analyses. The presence of specific IgG antibody against T. gondii in plasma was explored using ELISA method. DNA was extracted and conventional-PCR method was done using RE primers on buffy coat samples. Anti-Toxoplasma IgG were detected among 105/272 RA (38.6%) and 11/33 (33%) SLE patients (P=0.55). IgG seropositivity was more common in females (95.70%) than in males (4.30%) and in 51-60 years old patients (37.10%) in rural populations (62.07%). Also, seropositivity was higher in patients who kept cats (93.95%). Keeping cats and patient's age were two studied risk factors which had significant relations with T. gondii seropositivity in patients (P<0.05). Overall, T. gondii DNA was found in 60 of 305 (19.7%) of enrolled patients, whether serology positive or negative (P<0.0001). Given the RA and SLE patients can be considered as a risk group for toxoplasmosis, in addition to the serological test, PCR based techniques is recommended for early and accurate detection of recent T. gondii infection.

Global status of neglected human Linguatula infection: a systematic review of published case reports.

Human linguatulosis is a rare but highly specialized pentastomid parasitic infection all around the world. The disease is caused by worm-like parasites which share some morphologic features of arthropods and annelids. The life cycle of Linguatula spp. is provided as an example of pentastomid biology. The adult forms of Linguatula spp. are found in the nasal passages and frontal sinuses of dogs and other canids, domestic animals, and humans. Infestation may occur in the viscera, where immature forms develop in the liver, spleen, lungs, eyes, and other organs, or in the nasopharyngeal area. This review includes previous studies on the genus Linguatula in Iran and other countries from 1940 to 2019. Considering a few reports of human infestation, a comprehensive search was performed on both English and Persian databases for publications reporting human cases all around the world. Finally, 30 papers of 62 human cases were included in this study. The mean age of the patients was 25.8 years (varied from 8 to 79 years). Altogether, 41 (66.1%) and 21 (33.9%) of the patients were female and male respectively. The maximum cases (n = 25) were from Sudan. Nasopharynx was the most frequently involved organ in human cases. Our findings offer that physicians should consider the Linguatula infection in patients with complaints like unexpected dysphagia, with coughing, sneezing, and vomiting, after consuming raw/undercooked contaminated ruminant's viscera.

Alkaline phosphatase and score of HRCT as indicators for predicting the severity of COVID-19.

BACKGROUND: The clinical symptoms, blood laboratory data, O2 saturation and high-resolution computed tomography (HRCT) findings are critical factors in diagnosis of COVID-19 infection. METHODS: In this study, 105 hospitalized patients suspected of having COVID-19 were evaluated. Finally, the laboratory and HRCT and related factors data of 83 confirmed cases by HRCT and RT-PCR were analyzed. To compare the median of quantitative variables in the two groups, the Mann-Whitney U test was used. Also, to determine the factors associated with the positiveness of the HRCT result, a univariate logistic model was fitted. Moreover, receiver operating characteristic (ROC) curves were constructed to test the ability of the final model to predict the positiveness of HRCT result. RESULTS: 61.40% of the patients had a comorbidity disease. 89.20% had fever, 92.00% cough, 91.40% dyspnea. Abnormal CRP was seen in 77.80% of the patients, followed by 66.70% lymphopenia, and 60.30% neutrophilia. Also, ALP (abnormal vs. normal) and score of HRCT assessment variables had a significant effect on the positiveness of HRCT findings. 87.95% had abnormal HRCT with 41% bilateral multi lobar patchy ground glass opacity (GGO). Moreover, there was a statistically significant association between the level of O2 saturation and HRCT results. CONCLUSION: Our findings showed that male patients with middle age and comorbidity disease were more susceptible to the COVID-19 infection. Additionally, clinical features, blood laboratory findings, O2 saturation and HRCT findings are critical factors in the prognosis of COVID-19 infection.

Dietary Patterns Are Associated with Risk of Diabetes Type 2: A Population-Based Case-Control Study.

OBJECTIVE: We aimed to assess the relationship between major dietary patterns and risk of diabetes type 2 among Iranian adults. METHODS: In this population-based case-control study in Mazandaran province, we enrolled 332 subjects (110 newly diagnosed cases and 222 controls) aged 43 - 77 years. Dietary intakes were collected using a validated semi-quantitative food frequency questionnaire (FFQ). Data on demographic, anthropometric, socioeconomic characteristics and other covariates were collected using structured lifestyle questionnaires. Factor analysis was used to identify major dietary patterns. Odds ratios (OR) were calculated for risk of type 2 diabetes across quartiles of dietary pattern scores. RESULTS: Three major dietary patterns were identified, including: "healthy", "transitional" and "traditional". A significant direct association was found between the transitional dietary pattern and risk of diabetes type 2 after adjustment of potential confounders (OR = 2.17; 95% CI: 1.0, 4.50; Ptrend = 0.02). The traditional dietary pattern was significantly associated with the increased risk of diabetes type 2 after controlling for confounders (OR = 2.13; 95% CI: 1.03, 4.41; Ptrend = 0.01). There was no significant relationship between healthy dietary pattern and risk of diabetes type 2. CONCLUSIONS: In conclusion, transitional dietary pattern characterized by high consumption of salt, organ meats, dried fruits, poultry, tea, low- fat dairy and other vegetables. Traditional dietary pattern characterized by high intakes of garlic, dough, high- fat dairy, dried fruits, red meats, grains, as well as animal and hydrogenated fats were associated with an increased risk of type 2 diabetes. No significant associations were found between the healthy dietary pattern and risk of diabetes type 2.

The prognostic effect of PTEN expression status in colorectal cancer development and evaluation of factors affecting it: miR-21 and promoter methylation.

BACKGROUND: PTEN is a tumor suppressor gene which is involved in cellular proliferation, differentiation, and apoptosis. Loss or down-regulation of PTEN plays an important role in human cancers development. In this study, we investigated the effect of miR-21 and promoter methylation on the PTEN expression status in CRC tissues and analyzed association of the PTEN expression status with clinicopathological features in patients with CRC. RESULTS: The PTEN expression was positively detected in 67.2 % CRC tissues and all adjacent non-cancerous samples. PTEN mRNA level was negatively correlated with miR-21 level (r = -0.595, P < 0.001). PTEN expression was also correlated directly with the PTEN mRNA level (r = 0.583, P < 0.001) and conversely with miR-21 level (r = -0.632, P < 0.001). PTEN Promoter methylation was significantly associated with PTEN expression status (p = 0.013). PTEN expression was negatively associated with tumor size (p = 0.007) and advanced tumor stage (P = 0.011). Multivariate analysis indicated that tumor stage, tumor differentiation and PTEN expression status were independent prognostic factors for overall carcinoma in CRC patients (P < 0.05). The Kaplan-Meier curve indicated a negative correlation between PTEN expression levels and survival of CRC patients (P = 0.013). CONCLUSIONS: This study suggests a high frequency of miR-21 overexpression and aberrant promoter methylation in down-regulation of PTEN expression in colorectal carcinoma. Loss of PTEN may be a prognostic factor for patients with CRC.

Promoter methylation analysis of WNT/ß-catenin pathway regulators and its association with expression of DNMT1 enzyme in colorectal cancer.

BACKGROUND: Aberrant DNA methylation as the most important reason making epigenetic silencing of genes is a main mechanism of gene inactivation in patients with colorectal cancer. In this study, we decided to identify promoter methylation status of ten genes encoding WNT negative regulators, and measure the expression of DNMT1 enzyme in colorectal cancer samples. RESULTS: Aberrant methylation of APC gene was statistically significant associated with age over 50 (p = 0.017), DDK3 with male (p < 0.0001), SFRP4, WIF1, and WNT5a with increasing tumor stage (p = 0.004, p = 0.029, and p = 0.004), SFRP4 and WIF1 with tumor differentiation (p = 0.009 and p = 0.031) and SFRP2 and SFRP5 with histological type (p = 0.001 and p = 0.025). The increasing number of methylated genes correlated with the expression levels of the DNMT1 mRNA. CONCLUSIONS: The rate of gene promoter methylation of WNT pathway regulators is high in colorectal cancer cells. Hyper-methylation is associated with increased expression of the DNMT1 enzyme.

A rapid and reliable genotyping method for hepatitis B virus genotypes (A-H) using type-specific primers.

Eight genotypes of HBV (A-H) are recognized. A simple, rapid, and more specific genotyping system for HBV involving PCR using type-specific primers is described. The complete genomes of 234 human HBV strains for all the genotypes submitted to GenBank were aligned. The type-specific primers were designed based on the differences in the sizes of bands for eight genotypes in two sets. This genotyping system was tested with 24 positive HBV DNA controls. PCR was performed using two sets of type-specific primers for each sample in two tube. All 24 samples were PCR positive and possessed type-specific bands. PCR mix containing set 1 primers revealed specific bands of genotypes B, C, F and G, whereas PCR mix containing set 2 primers revealed specific bands of genotypes A, D, E and H. Type-specific PCR products were identified accurately by their sizes in agarose gels. The simplicity and rapidity of this PCR assay may reduce the cost and complexity of recognizing these genotypes. This method may be useful for HBV genotyping in large-scale clinical and epidemiologic studies.

Use of integrase-minus lentiviral vector for transient expression.

OBJECTIVE: Lentivirus-derived vectors are among the most promising viral vectors for gene therapy which is currently available, but their use in clinical practice is limited due to associated risk of insertional mutagenesis. Gene targeting is an ideal method for gene therapy, but it has low efficiency in comparison to viral vector methods. In this study, we are going to design and construct an integrase-minus lentiviral vector. This vector is suitable for transient expression of gene and gene targeting with viral vector. MATERIALS AND METHODS: In this experimental study, three missense mutations were induced in the catalytic domain of Integrase gene in the pLP1 plasmid and resulted D64V, D116A and E152G changes in the amino acid sequence through site directed mutagenesis. The pLenti6.2-GW/EmGFP transfer vector, associated with native and mutated packaging mix, was transfected into 293T cell line. In order to titer the lentivirus stock, the viruses were harvested. Finally, the viruses transduced into COS-7 cell line to assess green fluorescent protein (GFP) gene expression by a fluorescence microscopy. RESULTS: Recombinant and wild lentiviruses titer was about 5~8×10(6) transducing units/ml in COS-7 cell line. The number of GFP-positive cells transduced with native viruses was decreased slightly during two weeks after viral transduction. In contrast, in the case of integrase-minus viruses, a dramatic decrease in the number of GFP positive cells was observed. CONCLUSION: This study was conducted to overcome the integration of lentiviral genome into a host genome. Nonintegrating lentiviral vectors can be used for transient gene expression and gene targeting if a Target gene cassette is placed in the lentivirus gene structure. This combination method decreases disadvantages of both processes, such as random integration of lentiviruses and low efficiency of gene targeting.