Objective To establish the expression of membrane-bound HLA-G1-G4 isoforms in choriocarcinoma cell line JAR and to investigate its roles in NK cytotoxicity in vitro. Methods Stable expression of HLA-G1, -G2, -G3 and -G4 in JAR cells was established by gene cloning and transfection.HLA-Gtranscripts and protein expression in the transfected JAR cells was tested by RT-PCR, flow cytometry, Western blot and immunocytochemistry, respectively. High-affinity peptide KIPAQFYIL pulsing was performed to evaluate its effects on HLA-G expression. Effects of HLA-G1-G4 isoforms on NK cytotoxicity was performed with lactic dehydrogenase (LDH) releasing method. Results RT-PCR, Western blot and immunocytochemistry results showed that exogenous HLA-G1-G4 gene were successfully transfected and proteins were stably expressed in the HLA-G negative JAR cells; Flow cytometry data showed that only HLAG1, but not HLA-G2-G4 isoform was detectable in those transfected JAR cells and the peptide pulsing did not affect their expression status. However, all HLA-G1-G4 isoform expressed JAR cells could significantly decreased the NK cell cytotoxicity (P＜0.05). Conclusion HLA-G1-G4 isoform expression could dramatically inhibit NK-92 cell lysis, indicating that membrane-bound HLA-G isoforms are importantly immunotolerant and may play immune regulation roles in various physio-pathological situations.

Objective To investigate clinical effect of breviscapine combined with nerve growth factor in treatment of ischemic optic neuropathy . Methods 80 patients with ischemic optic neuropathy from March 2014 to March 2016 in our hospital were selected and divided into the control group and observation group, 40 cases in each group.The control group was treated with nerve growth factor, hip injection 30ug, one times a day, while the observation group was treated with breviscapine combined with nerve growth factor, 80 mg breviscapine added to 500 mL glucose intravenous infusion, one times a day.Then the changes of visual acuity, visual field loss difference (LV), the average damage of vision (MD), the average vision light sensitivity ( MS ) , adverse reactions, curative effect were compared between two groups.Results After treatment, the total effective rate in the observation group was significantly higher than the control group, the difference was statistically significant (P<0.05), the visual acuity of the observation group was significantly higher than that of the control group, the difference was statistically significant (P<0.05), LV, MD was significantly lower than the control group, the difference was statistically significant (P<0.05), MS was significantly higher than the control group, the difference was statistically significant (P<0.05).There was no difference in adverse reactions between the two groups.Conclusion Breviscapine combined with nerve growth factor in the treatment of ischemic optic neuropathy can improve nerve function of patients , improve the therapeutic effect.

Objective To investigate the effect and clinical efficacy of pranoprofen eye drops on lysozyme, EGF and LF levels in type 2 diabetic patients with dry eye syndrome.Methods70 Type 2 diabetes patients with dry eye from department of ophthalmology, Yuyao People's Hospital of Zhejiang Province from September 2013 to May 2015.They were selected and randomly divided into the treatment group and the control group with 35 cases in each group.The control group were treated by artificial tears eye drops 0.1mL/time, 6 times/day treatment.The treatment group were treated on the base of the control group with pranoprofen eye drops 0.15mL/eye drops, 4 times/day treatment.2 groups were 14 days for 1 courses of treatment, a total of 2 courses of treatment.The tear break-up time, basal lacrimal secretion, corneal fluorescein staining positive rate and clinical efficacy were compared between two groups before and after treatment.ResultsWith the total effective rate in the treatment group,was higher than the control group(P<0.05).Compared with before treatment, the tear break-up time prolonged, basal lacrimal secretion, tear lysozyme, EGF and LF levels were higher(P<0.05), the positive reaction rate of corneal fluorescence staining was lower(P<0.05).Compared with the control group after treatment, the tear break-up time prolonged, basal lacrimal secretion, tear lysozyme, EGF and LF levels were higher(P<0.05), the positive reaction rate of corneal fluorescence staining was lower(P<0.05).ConclusionThe clinical efficacy of pranoprofen eye drops in type 2 diabetic patients with dry eye syndromeis good.It is speculated that the mechanism and tear break-up time prolonged, basal lacrimal secretion increase and lysozyme, epidermal growth factor(EGF)and the level of LF increased.

Objective: To discuss the applied effect of high frequency repetitive transcranial magnetic stimulation combined with cognitive intervention for stroke patients with aphasia. Methods: 84 stroke patients with aphasia were divided into observation group (42 cases)and control group (42 cases). Patients of control group were treated by routine intervention of language training, while patients of observation group were treated by high frequency repetitive transcranial magnetic stimulation combined with cognitive intervention. A series of evaluated indicators, such as the language function score of Boston diagnostic aphasia examination(BDAE), the anxiety symptom score of Hamilton anxiety scale(HAMA), the quality of life and degree of satisfaction, were adopted to compare the post-treatment effect for the two groups. Results: After the treatment, there were statistical significance in the difference of understanding, retelling ability, reading ability and writing ability about BDAE scores between the two groups (t=3.572, t=4.482, t=6.238, t=3.013; P<0.05), and the HAMA scores of observation group was significant higher than those of control group (t=3.472, P<0.05). Besides, through different interventions, the series of indicators about quality of life, such as physical function, mental function, social function and material function, of observation group were higher than those of control group (t=4.513, t=3.923, t=4.473, t=4.033; P<0.05). And the satisfaction of observation group was significant higher than that of control group (x2=5.233, P<0.05). Conclusion: The clinical effect of high frequency repetitive transcranial magnetic stimulation combined with cognitive intervention for stroke patients with aphasia is favorable. And this method can increase quality of life and satisfaction for patients, and can extremely reduce medical dispute and improve the therapeutic level of hospital for stroke patients with aphasia.

Objective To study the expression of endometrial integrin-? v? 3 in infertility patients induced by chronic pelvic inflammation disease(PID)and to analyze the correlation between the expression and the TCM syndrome patterns.Methods Twenty five infertility patients induced bychronic PID were enrolled into the experimental group,and 15 child-bearing aged women with normal reproductive capability were in the control group.The expression of integrin-? v? 3 in endometrium was measured by flow cytometry.The expression of integrin-? v? 3 in different TCM syndrome patterns was compared.Results The expression of integrin-? v? 3 was(4.76? 1.84)% in experimental group and(11.91? 1.04)% in the control group,there existed a significant difference(P

0.05),while that of serum P was significant(P

Objective To use the AU2700 biochemical analyzer to evaluate the performance of the reagent kit for determining se-rum glycated albumin(GA)by the enzymatic method.Methods On the basis of the relevant documents of CLSI-EP,the perform-ance evaluation of the Lucica GA-L reagent kit for detecting GA was performed on the precision,accuracy,linearity range and refer-ence range;the correlation between the results detected by the two kinds of reagent kit (enzymatic method)was analyzed.Results The reagent had high precision(CV<3%);the dilution had good linearity;the average recovery rate of GA was 97.7%,the average recovery rate of albumin(ALB)was 96.43%;the detection results by the two kinds of reagent kit had good correlation (r2 =0.976, P <0.01).Conclusion The Lucica GA-L reagent kit shows the good analytical performance on the AU2700 biochemical analyzer, which is consistent to the manufacturers statement and can be better used in clinic.

Objective To evaluate the dynamic changes of T lymphocyte subsets in children with hand-foot-and-mouth disease(HFMD)and to provide new evidence for the therapy and prognosis.Methods Peripheral venous blood samples of 346 HFMD cases in acute stage who were hospitalized in Beijing Ditan Hospital from May 1,2008 to August 31,2008 were collected and T lymphocyte subsets were assayed by flow cytometer.Meanwhile,T lymphocyte subsets of 67 HFMD cases in recovery phase were also detected.The pathogens were determined by reverse transcriptionpolymerase chain reaction(RT-PCR)using pharynx swab samples from 99 cases.Different samples were compared by independent-sample t test,paired t test or variance analysis.Results The average levels of T lymphocyte subsets of HFMD children in different agc groups were all lower than reference levels of healthy children in according age groups.In severe cases.T lymphocyte(TL)/lymphocyte (L)ratio in all age groups,helper T cell(Th)/L ratio in children older than 1 year,TL,Th and Th/suppressor T cell(Ts)ratio in children of 1-2 years old were all lower than those in common eases (P＜0.05).The Th/L ratio tended to increase with the disease progression.Ratios of TL/L and Th/L in common cases were increased in recovery phase(TL/L:56.3±8.6 vs 61.1±9.1,t=2.56,P＜0.05;Th/L:30.2±7.2 vs 34.9±7.9,t=2.90,P＜0.05)and all indices of severe cases except Ts/Lratio and Th/Ts ratio increased apparently in recovery phase(P＜0.01).TL[(1.738±0.976)×10~6/Lvs(2.696±1.946)×10~6/L,t=2.17,P＜0.05],Th/L ratio(25.9±7.0 vs 30.2±7.2,t=2.34,P＜0.05),Th[(0.864±0.550)×10~6/L vs(1.459±0.879)×10~6/L,t=2.90,P＜0.01]and L[(3.352±1.458)×10~6/L vs(4.664±2.435)×10~6/L,t=2.32,P＜0.05]of severe cases in acute phase were all lower than those of common cases(P＜0.05),while those were not significantly different in recovery phase between two groups(P＞0.05).The T lymphocyte subsets of enterovirus(EV)71 positive cases were lower than EVT1 negative cases,but there was no significant difference between these two groups(P＞0.05).Conclusion T lymphocyte immune responses may be correlated with HFMD onset and progression.

Human leukocyte antigen G ( HLA-G) is a member of the non-classical HLA classⅠb family. It is considered to play a crucial role in immune tolerance. A unique feature of HLA-G is the struc-tural diversity as surface expressed and as secreted molecules, which is mainly attributed to alternative spli-cing of the primary transcript. HLA-G can promote the invasion and metastasis of tumor cells through various ways. In addition, HLA-G has been described included in exosomes. Exosomes released by most cell types are nano-sized vesicular bodies that contain lipid bilayer and rich contents. As a new marker for diseases, exosomes are extensively involved in the occurrence and development of diseases. Recent studies have found that exosomes can express soluble HLA-G, which reveal a new way by which HLA-G regulates tumor micro-environment. In this review, we focus on the expression of HLA-G on exosomes to provide new thoughts for the early detection and treatment of tumors.

Objective To investigate the associations between single nucleotide polymorphisms in the gene encoding human leukocyte antigen-G ( HLA-G) and the genetic susceptibility to high risk human papillomavirus ( HPV) type 18 infection in the subjects from Taizhou, Zhejiang province. Methods The genetic polymorphisms of HLA-G gene (14 bp In/Del and +3142C/G) in cervical samples collected from HPV 18-positive and healthy women were analyzed by PCR and gene sequencing technology. Statistical anal-ysis was performed by using SPSS version 16. 0. Chi-squared test was used to analyze the differences with HLA-G gene allele and genotype frequencies between healthy subjects and patients. Results Compared with healthy subjects, women with oncogenic HPV18 infection showed lower frequencies of -14 bp allele,-14 bp/-14 bp genotype and -14 bp/+3142C haplotype (P<0. 05). Moreover, lower frequencies of+3142C/C genotype were detected in HPV18-infected women with pathologically normal cervix (6. 3% vs 21. 1%, OR=0. 25, P<0. 05) and higher percentages of +3142G/G genotype were detected in women with CIN2/3 stage HPV18 infection as compared with those of the control group (68. 8% vs 35. 1%, OR=4. 06, P<0. 05). Conclusion The HLA-G gene 3′ UTR polymorphisms were closely associated with HPV18 in-fection and cervical intraepithelial neoplasia.