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 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 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 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 investigate the mechanism of different HLA-G isoform mRNA patterns in different cells alters its cell membrane expression.Methods RT-PCR was used to analyze HLA-G isoform mRNA(HLA-Gl-6)of ovarian cancer cell lines HO-8910,HO-8910PM and OVCAR-3,leukemia cell lines Jurkat,K562,HIJ60,MUTIZ-1,and the chorioeareinoma cell lines JEG-3,JAR.HLA-G between cellular membrane and intracellular expression were analyzed by flow cytometry.Results All HLA-G mRNA isoforms were observed in the positive control cell line JEG-3,but none in the negative control cell line JAR.HLA-G1 isoform mRNA was expressed in HO-8910,HO-8910PM,OVCAR-3,MUTZ-1 and Jurkat cells.HLA-G2 mRNA was not detected in any cell line but JEG-3.HLA-G3 mRNA was found in HO-8910,HO-8910PM,K562,HIJ60,MUTZ-1,OVCAR-3 and Jurkat cells.HO-8910,HO-8910PM,HIJ60,Jurkat cells expre8sed HLA-G4 mRNA.Only the Jurkat cells expressed HLA-G5 mRNA.FACS results showed that JEG3 and HO-8910PM cells membrane expressed HLA-G,however,the intraeellular HLA-G expression was detected in all tested cells except the negative control cell JAR.Conclusion Only the HLA-G1 isoform could be exDressed on cell membrane in particular cell lines. Other isoforms including HLA-C2,-G3,-G4,-G5 and HLA-G6 could not reach cell snrface.

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.

Objective To investigate the security characteristics of rural young boarding pupils and its influencing factors.Methods 210 boarding and 265 non-boarding rural young pupils of Huojia in Henan were tested by questionnaires,including of a self-developed questionnaire,security questionnaire and adolescent mental resilience scale.Results ①The scores of the interpersonal security (25.99 ± 5.53),the determine control sense factor (24.63 ±6.17) and the total security(50.62 ± 10.37) in the boarding pupils were statistically lower than non-boarding pupils (27.36 ± 6.38,26.00 ± 6.55,53.36 ± 11.77,P ＜0.05).②There were statistically significant difference in the scores of the interpersonal security,the determine control sense factor and the total security among the different grades(P＜0.05),but there were no statistically sex difference in the scores(P ＞ 0.05).③Group regression analysis showed that,no family factors in the young boarding pupils had influence on the score of the security sense factors and the total (P＞0.05),and marital relationship of parents,only child,father' s and mother(')s education background of non-boarding pupils had certain influence on the score of the interpersonal security,the determine control sense factor and the total security (β =-0.43-0.27,P＜ 0.05).④The group regression analysis showed that,the problem solving and self-efficacy,parents'high expectations in mental toughness respectively had some effect on the interpersonal security of boarding and non-boarding pupils(β =-0.15,β =0.25,P＜0.05),and social ability and family care,schools resilience respectively had influence on the determine control sense in boarding and non-boarding pupils (β =-2.91-2.87,P＜0.05),and social ability,parents'high expectations respectively had influence on the total security in boarding and non-boarding pupils (β =-0.17,β=0.22,P＜0.05).Conclusion The security-sense in boarding young pupils are lower than the non-boarding young pupils,and family factors and mental toughness have different effects on security-sense of two kinds pupils.

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.