Immunotherapy is becoming an effective and less invasive strategy that can be applied to the treatment of various malignancies. Lentiviral vectors (LVs) have shown great potential in immunotherapy as they can stably integrate relatively large foreign DNA, and effectively transduce dividing and non-dividing cells. Clinical application needs high quality LVs, and therefore strict quality control of the final products is necessary to ensure their purity, efficacy and safety. The quantitative detection of LVs is among the key parts of product development and quality control. In this paper, the existing methods for quantitative detection of LVs are summarized, including fluorescence activated cell sorter (FACS), P24 enzyme-linked immuno sorbent assay (P24 ELISA), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), nanoparticle tracking analysis (NTA), tunable resistive pulse sensing(TRPS) and virus counter(VC).Their advantages and disadvantages are listed, and future development and challenges are discussed.
Genetic Vectors/genetics* ; Humans ; Immunotherapy ; Lentivirus/genetics* ; Neoplasms ; Transduction, Genetic

Objective To study the curative effect of the laser photocoagulation combined with Xuefu Zhuyu decoction in treatment of patients with ocular fundus disease. Methods Ninety-seven patients with ocular fundus diseases were selected from the Department of Ophthalmology, The Central Hospital of Lishui City from June 2016 to June 2017. The patients were randomly divided into control group and observation group by the odevity of their sickbed number, including 48 odd numbers in control group and 49 even numbers in observation group. The patients in control group were treated with laser photocoagulation. The observation group added Xuefu Zhuyu Decoction in the foundation of control group. We compared the curative effects, side effects and vision improvements of each group. Results The total effective rate of the observation group was 95. 92％, which was significantly higher than the rate of the control group at 83. 33％. There was significant difference between the two groups(P＜0. 05). The vision improvements of the observation group after three and six months were better than those of the control group and the difference between the two groups was significant(P＜0. 05). The total numbers of occurrences of side effects in the control group and the observation group were 4 cases(8. 33％) and 5 cases(10. 20％) respectively, while the difference wasn't significant(P＞0. 05). Conclusion The laser photocoagulation combined with Xuefu Zhuyu decoction in treatment of patients with ocular fundus disease has better curative effect and less side effects, which helps visual recovery effectively, deserving clinical application and popularization.

Objective:To study the effects of compound schisandra extracts (CSE) (schisandra,astragalus,acanthopanax,and rhodiola)on the exhaustive swimming time and the levels of blood urea nitrogen(BUN),serum lactic acid(LD),liver glycogen and muscle glycogene,superoxide dismutase(SOD)activity,and malonaldehyde(MDA) level in the mice and to charify its anti-fatigue effect and the mechanism.Methods:Eighty male ICR mice were randomly divided into blank control group,50 mg·kg-1 CSE group,100 mg·kg-1 CSE group,and 200 mg·kg-1CSE group;there were 20 mice in each group.The mice were administered orally for 30 d.Then 10 mice were randomly selected for exhaustive swimming test in each group and the exhaustive swimming time of the mice was recorded.The remaining 10 mice in each group were used for 90 min swimming,then all the mice were sacrificed and the blood and tissue samples were taken for the measurement of the levels of BUN,LD,liver glycogen and muscle glycogen,the SOD activity and MDA level;the total inhibitory rate of oxidation of CSE in vitro was determined by linoleic acid-ferric thiocyanate method.Results:Compared with blank control group,the exhaustive swimming time of the mice in 50,100,and 200 mg·kg-1 CSE groups were significantly increased (P<0.01);the levels of BUN and LD of the mice in 100 and 200 mg·kg-1 CSE groups were significantly decreased (P<0.05 or P<0.01);the levels of liver glycogen and muscle glycogen of the mice in 100 and 200 mg·kg-1 groups were significantly increased(P<0.05 or P<0.01);whereas the SOD activities were significantly increased and the levels of MDA were decreased(P<0.05 or P<0.01).Compared with 100 mg·kg-1 CSE group,the levels of serum BUN and LD of the mice in 200 mg·kg-1 CSE group were decreased (P<0.01),and the levels of liver glycogen and muscle glycogen were increased(P<0.05).The total inhibitory rate of oxidation of 5 g·L-1CSE was 76.94%.Conclusion:CSE has an anti-fatigue effect and the mechanism may be related to anti-oxidation effect.

Objective:To study the effect of breviscapine on visual function of the patients with primary open-angle glaucoma after filtration surgery. Methods:Twenty-nine cases (42 eyes) of patients with primary open-angle glaucoma at middle and late stage after filtration surgery were selected and randomly divided into the experimental group and the control group according to the defect degree of visual field. The 15 patients (21 eyes) in the experimengtal group ( TG) were intravenously injected 250ml saline containing 50mg breviscapine, while the 14 patients (21 eyes) in the control group (CG) were intravenously injected 250ml saline containing 1g vita-min C. The treatment course was 2 weeks followed by one-week withdrawal. After three treatment courses, the visual function in the two groups was investigated and compared before and after the treatment. Results:After the treatment, the visual field defect value and the thickness of retinal nerve fiber layer of the patients in TG were both better than those before the treatment (P<0. 01,P<0. 05). The values of visual acuity and visual field defect of the patients in TG were significantly better than those of the patients in CG ( P<0. 05, P<0. 01). Conclusion:After intraocular pressure is controlled, breviscapine injection can reduce the vision damage of patients with primary open-angle glaucoma at middle and late stage after filtration surgery and improve the visual field defect.

Objective To investigate the prognosis value of pulmonary infection score and acute physiology and chronic health evaluation to acute exacerbation of COPD with type 2 respiratory failure. Methods 112 cases with acute exacer-bation of COPD with type 2 respiratory failure were selected as research subjects in our hospital from February 2011 to February 2014,scores of two scoring methods,actual mortality with different scores of two scoring methods,ROC curve predicted mortality of two scoring methods to chronic obstructive pulmonary disease with acute exacerbation of respira-tory failure of survival group and death group were compared. Results There were no significant differences between the two groups in PaO2、pH、PaCO2 and HCO3-(t=1.742, 1.647, 1.791, 1.082, P>0.05);but the levels of AG and poten-tial HCO3-in death group were higher than those in survival group(t=10.865, 5.993, P<0.05). APACHEⅢ,CPIS scores of survival group significantly were lower than the death group, the differences were statistically significant (t=6.254,4.397,P<0.05). Mortality of APACHⅢ score ≥25 and CPIS score ≥6 were significantly higher than patients with lower than that value, the differences were statistically significant(χ2=10.264,7.351,P<0.05). CPIS predict mortality acute exacerbation of COPD with type 2 respiratory failure, the area under the ROC curve was 0.867,APACHEⅢpredict mortality acute exacerbation of COPD with type 2 respiratory failure, the area under the ROC curve was 0.938. Conclusion CPIS and APACHEⅢ has certain prognostic value to acute exacerbation of COPD and type 2 respiratory failure,but APACHEⅢ resolution are better, it can provide a reference for disease prediction, it is worthy of applica-tion and promotion in clinical.

BACKGROUND:Adult cardiomyocytes show no regenerative ability, and celltherapy for myocardial regeneration and repair may improve myocardial ischemic injury function. OBJECTIVE:To confirm the effect and reveal the mechanism of bone marrow mesenchymal stem cells (BMSCs) on acute myocardial infarction (AMI). METHODS:BMSCs were isolated, cultured from bone marrow of Sprague-Dawley rats using density gradient centrifugation. AMI models were produced in 20 rats by ligating the left anterior descending (LAD) coronary artery, and randomly divided into model group and BMSCs group. In the BMSCs group, cells were subsequently injected with a sterile microinjection via the tail vein. RESULTS AND CONCLUSION:Six months postoperatively, the cardiac function was improved, the vessel density was increased, the percentage of apoptotic cells was decreased in the BMSCs group than that in the model group;the expression levels of inflammatory factors, including vascular endothelial growth factor, von Wil ebrand factor, transforming growth factor 3β, and interleukin-1βmRNA were significantly improved in the BMSCs group than that in the model group. These results showed that BMSCs can protect the myocardium from AMI by regulating the secretion of inflammatory cytokines and angiogenic factors.

Objective To detect Pneumocystis carinii in sputum in patients with AIDS complicated with Pneumocystis pneumoniae (PCP)by using indirect immunofluorescence microscopy.Methods Indirect immunofluorescence microscopy was used to detect Pneumocystis carinii in sputum samples of 53 patients with AIDS complicated with suspected PCP,observed the clinical effects and outcomes of the patients.Results Of the 53 cases,30 cases were positive.The positive rate was 56.6％.Patients with positive CD4+ T lymphocyte count was in average 33.33 ± 42.99/μl,23 patients who were negative for PCP had a CD4+ T lymphocyte count of 104.13 ± 77.87/μ1.Conclusions The indirect immunofluorescence microscopy was specific rapid and sensitive method in detection of pneumocystis infection in HIV infected patients.

Objective To explore the effect of BCG-polysaccharide nucleic acid(BCG-PSN) on the numbers of Th3,Th2 and Th1 cells in peripheral blood mononuclear cell(PBMC) and airway inflammation status in patients with asthma.Methods Twenty-six patients with moderate persistent asthma were enrolled into the study and randomly divided into simple medication group(n=13,accepted Fluticasone/salmeterol alone) and combined medication group(n=13,accepted Fluticasone/salmeterol+BCG-PSN together).The numbers of Th3,Th1 and Th2 cells in PBMC, the serum levels of TGF-β,IFN-γ,IL-4 and IgE and the clinical effect and airway inflammation status were observed at three time points: before,1.5 months after and 3 months after treatment. Results (1)At 1.5 months after treatment,the Th1/Th2 number was significantly higher in the combined medication group than the simple medication group(1.20±0.57 vs 0.79±0.39,t= 2.129,P＜0.05),and the asthma control test scale(ACT) showed similar difference(18.31±1.75 vs.15.54±2.40,t=3.359,P＜0.01) between the two groups.(2)At 3 months after treatment,the Th1/Th2 number was significantly higher in the combined medication group than the simple medication group(1.73±0.74 vs1.16±0.48,t=2.327,P＜0.05),and the ACT also showed the same kind of difference(22.46±1.13 vs.20.23±2.59,t=2.851,P＜0.01) between the two groups.In addition,at this time point the PEF variability was significant lower in the combined medication group than the simple medication group([9.88±2.18]% vs.[12.05±2.74]%,t=2.235,P＜0.05).(3)We found no significant differences in the comparisons of the numbers of T helper cell subsets in PBMC ,the serum levels of cytokines and IgE,the numbers of inflammatory cells in induced sputum between the two groups(Ps＞0.05).(4)No correlation were found between Th3 cell numbers in PBMC and the serum levels of IgE in the two groups.Conclusion The combined use of Fluticasone/salmeterol and BCG-PSN can further correct the imbalance of Th1/Th2 cells,alleviate clinical symptom of asthma and airway hyper-responsiveness.The therapeutic efficacy improves along with the therapy period,better in the combined medication group than the simple medication group.BCG-PSN has no significant effects on the numbers of Th3 in PBMC in patients with asthma.

ObjectiveTo investigate the expression of T lymphocytes and activated subsets in patients with HIV/AIDS and early latent syphilis. MethodsT lymphocytes and activated subsets ( HLADR+ CD3 +/CD3 + , HLA-DR+ CD4 +/CD4 + and HLA-DR+ CD8 +/CD8 + ) as well as rapid plasma reagin (RPR) and treponema pallidum particle agglutination (TPPA) test were detected by flow cytometry in 78 patients with HIV/AIDS, 66 patients with HIV/AIDS and early latent syphilis, and 30 healthy subjects. SPSS 13.0 was used for statistical analysis, and t (for normal distribution) or Mann-Whitney U (for skew distribution) tests were performed to compare between the groups. ResultsThe absolute counts of CD4+ T cells in patients with HIV/AIDS and early latent syphilis were significantly higher than those in HIV/AIDS patients ( t = 2. 041 and 2. 223, P ＜ 0.05 ), but no difference in the counts of CD3 + T cells and CD8 + T cells was observed (tcD3 =0. 362 and 0.692, tcD8 =0.043 and 0.617, P＞0.05). HLA-DR+ CD4 +/CD4 +level in AIDS plus syphilis group was much higher than that in HIV plus syphilis group ( t = 2. 647, P ＜ 0. 05 ), but no difference was observed in HLA-DR+ CD3 +/CD3 + and HLA-DR+ CDs +/CDs + ( t = 1. 112 and 0. 093, P ＞ 0.05). ConclusionsImmune function in patients with HIV/AIDS and early latent syphilis may be enhanced temporarily.

ObjectiveTo study the clinical features and pathogenesis of ophthalmoplegia in senile patients with diabetes, and access the possibility of its development.MethodsBesides routine ophthalmologic and ocular muscle examination, general and laboratory examinations were also done. Some patients were diagnosed also with the help of image processing.ResultsMonocular involvement happened in all 21 patients. 12 patients suffered from oculomotor nerve paralysis, 5 abducent nerve paralysis, 3 trochlearis nerve paralysis, and 1 compound nerve paralysis. Among these patients with complete oculomotor nerve panalysis, no papillary change appeared.Conclusions Microvascular pathologic change may be the pathogenetic basis of ophthalmoplegia for senile diabetic patients. Its prognosis is favorable if reasonable treatment is made.