0.05).Conclusions With the increase of the ejaculatory frequency,human semen volumes and the total counts decrease,while sperm viability and motility increase.Successive two ejaculation does not affect the quality of human semen.

Objective: To investigate the mechanisms of anti-tumor cytotoxicity of human ??T cells activated by Mycobacterium tuberculosis antigen (Mtb-Ag) . Methods: The peripheral blood mononuclear cells (PBMC) of healthy donors were stimulated by Mtb-Ag in vitro to activate preferentially ??T cells that were expanded in IL-2 contained medium.The high purified ??T cells were isolated by the positive selection on MACS separator. The expression of mRNA of perforin, granzyme B and Fas Ligand in purified ??T cells were measured by RT-PCR technique. Fas Ligand on ??T cells and Fas molecule on tumor cells were detected by flow cytometry. The MTT assay was used to measureanti-tumor cytotoxicity of ??T cells. The inhibitory effects of concanamycin A (CMA) for perforin/granzyme pathway, and brefeldin A (BFA) for Fas/Fas ligand pathway on cytotoxicity of ??T cells were observed. Results: The mRNA of perforin and granzyme B and Fas ligand were highly expressed in purified ??T cells. The cytotoxicity of ??T cells against K562 and PG that expressed low level of Fas molecule (6.5% and 7.8%) was remarkably inhibited by pretreatment of the ??T cells with CMA (200 nmol/L) by 72% and 76%, respectively, while the BFA(30 ?mol/L) showed less inhibitory effects by 32% and 25%, respectively; However, it was showed that the BFA (30 ?mol/L) preferentially inhibited the cytotoxicity of ??T cells against tumor cell lines Raji and A375 that expressed high level of Fas molecule on cell surface (98.5% and 70.6%, respectively).Conclusion: Preferentially, granule exocytosis was a main mechanism by which the Mtb-Ag activated ??T cells showed cytotoxicity aganist tumor cells that expressed low level of Fas molecule, whereas both Fas/Fas ligand pathway and granule exocytosis were involved in the cytotoxicicty of Mtb-Ag activated ??T cells against the tumor cells that expressed high level of Fas molecule.

Objective To detect the frequency of peripheral blood CD4+ CD25high Tr and CD4+CD25lowT cells and expression of programmed death-1 ( PD-1 ) on their surface in the patients with systemic lupus erythematosus(SLE) and their clinical signifcance is analyzed.Methods The expression of PD-1 on the CD4+ CD25highTr and CD4+CD25lowT cells was examined in patients with 33 active SLE,18 inactive SLE and 38 healthy controls(HC) by flow cytometry.Clinical manifestations and laboratory findings of SLE were collected.Patients were divided into two groups according to their disease activity.SLE disease activity index(SLEDAI) score≥10 was defined as high disease activity and ＜10 as inactivity.The percentage of CD4+ CD25highTr and CD4+CD25lowT cells and proportions of expression of PD-1 on their surface were compared between not only inactive or active SLE patients and healthy controls (HC),but also between patients with lupus nephritis and without lupus nephritis.Correlation with clincal manifestations and laboratory findings was analyzed.Results (1)The proportions of CD4+CD25highTr and CD4+CD25low T cells were significantly decreased in active and inactive SLE patients as compared with HC.(2)The percentage of CD4+ CD25lowPD-1 +Tr and in active and inactive SLE patients were higher than HC.The proportions of CD4+CD25lowPD-1+ T cells were significantly increased in HC and inactive SLE patients as compared with inactive SLE patients.(3) The proportions of CD4+CD25highPD-1+Tr in SLE patients with nephritis were higher than those in patients without nephritis.But the percentage of CD4+ CD25 lowPD-1 + T cells was significantly decreased in SLE patients with nephritis as compared those without nephritis.(4)A positive correlation was observed for proportions of CD4+CD25highPD-1 + T cells with SLEDAI score.Proportions of CD4+ CD25high Tr,CD4+ CD25low PD-1 + T cells was inversely correlated with SLEDAI score.Conclusion The aberrations of proportions of CD4+CD25highpD-1+,CD4+CD25lowPD-1+,CD4+CD25high and CD4+CD25low T cells were observed in patients with SLE.The abnormality of PD-1 and PD-L1 pathway may play an important role in the function and number of Tr.

Objective:To observe the stimulating effect of C main Peptide purified from Mycobacterium tuberculosis peptide antigens(Mtb Ag) on human ??T cells, and the effect of C main peptide on the Mtb Ag activated ??T cells in vitro. Methods:C main peptide was used to stimulate fresh ??T cells of normal subjects in different doses for 10 d and the responded cells were phenotyped by flow cytometry. Furthermore,C main peptide was used to restimulate the activated ??T cells ,and then analyse the expression of early activation marker molecule CD69 in ??T cells by flow cytometry and the activity of proliferation of ??T cells by MTT assay. Results:C main peptide could stimulate the proliferation of fresh ??T cells predominantly, and it also could promote the expression of CD69 in activated ??T cells and enhance its activity of proliferation. Conclusion:C main peptide purified from Mtb Ag is a kind of specific stimulator of human ??T cell in vitro.

Objective The aim of this study is to examine the expressions of Toll like receptor (TLR) 7 and TLR9 in the peripheral blood B lymphocytes of SLE patients and to analyze the correlation between TLR7/9 and clinical parameters. Methods lntracellular expression of TLR7/9 in the peripheral blood CD19+Blymphocytes was analyzed in 50 SLE patients and 30 healthy controls by flow cytometry. The difference of intracellular TLR7/9 expression levels in two groups was compared. Furthermore,the correlation between TLR7/9 expression and clinical parameters such as ESR, CRP, complement 3 (C3), complement 4 (CA), the level of serum IgG, anti-double stranded DNA antibody, anti-nuclear antibodies, SLEDAI score and urine protein excretion level, were analyzed. Results Compared with healthy subjects, the proportion of B cells expressing TLR7 and TLR9 was higher among SLE patients. Positive correlation was observed between TLR7 expression levels and clinical measurement of the SLEDAI and ESR. Negative correlation was observed between TLR7 expression levels and serum C3 levels. Positive correlation was observed between TLR9 expression levels and SLEDAI scores. Negative correlation was observed between TLR9 expression levels and serum C3 levels. Conclusion TLR7 and TLR9 expression is increased in the peripheral blood B cells of SLE patients, and correlates well with clinical parameters.

Objective To explore the roles of PKCθ(protein kinase Cθ)signaling pathway on the activation,proliferation and TH1/TH2 cytokines production of the γδT lymphocytes stimulated by Mycobacterium tuberculosis antigen(Mtb-Ag) in vitro.Methods Peripheral blood mononuclear cells(PBMC)were pretreated with or without Rottlerin at 5.0 μmol/L,and stimulated with Mtb-Ag and cultured in rhIL-2 containing medium.After different time of culture,activation molecules and cytokines production of γδT cells were measured by flow cytometry.The proliferation proportion and the percentage of each generation of γδT cells were determined by carboxylfluoreseein diacetate succinmidyl ester(CFSE)labeling technique and flow cytometry.Results After 3d of stimulation with Mtb-Ag,the expression rates of CD69 and CD25 of γδT cells were 46.2%and 45.6%,respectively.Pretreatment of 5.0 μmol/L Rottlerin markedly inhibited the both expressions of CD69 and CD25 in γδT cells(P＜0.01).After stimulation and expansion in vitro for 5,10,and 15 d,the percentages of the γδT cell were 9.6%,54.6%and 82.4%,respectively.There was a few γδT cells in propagation on the 5th day of culture,and almost all γδT cell divisions were above 6 generations on the 10th and 15th day of culture.Pretreatment of the Rotflerin significantly suppressed the γδT cell proliferation,but after 10 d culture,there were still a few parts of γδT cells in propagation.Meanwhile,after 7,14,and 21d of culture,upon stimulation with PMA+Ionomycin,the IFN-γ producing-γδT cells were about 80%at all times.But only after 21d culture,IL-4-producing γδT cells was 2.6%.,The percentage of IFN-γ producing γδT cells markedly reduced in Rottlerin group(P＜0.01).IL-4 secretion of the γδT cells was almost completely blocked.Conclusion PKCO signal pathway involves in activation,proliferation and differentiation of the γδT lymphocytes stimulated by Mtb-Ag.

Objective To investigate the clinical significance of abnormally expressed PD-1 on CD 4+CD 2 8+/-T cells in peripheral blood of patients with systemic lupus erythematosus ( SLE ) . Methods Peripheral blood samples were collected form 50 patients with primary SLE and 40 healthy subjects and used to isolated mononuclear cells. Expression of CD4+CD28-, CD4+CD28+, CD4+CD28+PD-1+and CD4+CD28-PD-1+T cells in peripheral blood samples of the two groups were detected by flow cytometry. Clinical data of SLE patients were collected. Based on SLE disease activity index (SLEDAI), SLE patients were classified into two groups: stable group (SLEDAI<10) and active group (SLEDAI≥10). Based on the condition of renal damage, they were also divided into two groups: lupus nephritis group and non-lupus ne-phritis group. Differences in T cell expression were compared among these groups. Statistical analysis was performed to analyze the relationships of different T cell subsets with laboratory and clinical parameters rela-ting to SLE and SLEDAI. Results The percentages of peripheral CD4+CD28-, CD4+CD28+PD-1+and CD4+CD28-PD-1+T cells of active group were higher than those of stable and healthy control groups ( P<0. 05). Moreover, patients with lupus nephritis had higher percentages of these T cell subsets than those without (P<0. 01). SLE patients who were positive for anti-dsDNA or anti-SmRNP antibody, or had de-creased complement C3, thrombocytopenia or decreased lymphocytes had higher percentages peripheral CD4+CD28-T cells than those in the corresponding negative group. SLE patients who were positive for anti-dsDNA or anti-SmRNP antibody, or had decreased complement C3, complement C4 or lymphocytes showed en-hanced expression of peripheral CD4+CD28+PD-1+T cells as compared with those in the corresponding nega-tive group. SLE patients positive for anti-dsDNA antibody, or with decreased complement C3 or lymphocytes or suffering from alopecia had higher percentages of peripheral CD4+CD28-PD-1+T cell than those in the cor-responding negative group. Differences between different groups were statistically significant (P<0. 05). Conclusion Abnormal expression of CD4+CD28-T cells and PD-1 on CD4+CD28-and CD4+CD28+T cells in peripheral blood of patients with SLE has certain correlation with laboratory parameters and clinical indicators.

OBJECTIVETo understand the demographic characteristics of the HIV positive cases detected in the screening project in communicable disease prevention demonstration area in Guangxi Zhuang autonomous region and analyses the related factors of HIV infection.

METHODSHIV antibody detection was conducted for local people aged no less than 15 years old in 3 demonstration counties, and the demographic information of the subjects were collected. The factors influencing HIV infection were analyzed with software PASW Statistics 18.0.

RESULTSA total of 770 454 local people aged over 15 years were screened, the HIV infection rate was 1.58‰ among them. The positive cases were mainly married males aged≥30 years and with the educational level of primary school and junior middle school. Thus male, age over 30 years and educational level of primary school and junior middle school. were the common risk factors for HIV infection, whereas being married was a protective factor for HIV infection in 3 demonstration counties, but Han ethnic group and divorced/widowed were risk factors for HIV infection in the demonstration county A, non-farmer, divorced/widowed and educational level of senior middle school were the risk factors for HIV infection in demonstration county B and Han ethnic group and farmer were risk factors HIV infection in demonstration county C.

CONCLUSIONIt suggested to conduct targeted HIV infection screening according to the survey results to find out the HIV cases in general population.

Objective: Tracking the information on 1.69 million fetal cases across Guangxi Zhuang Autonomous Region (Guangxi) so as to study the occurrences of total and major birth defects in order to evaluate the ability on related prevention and control programs in Guangxi. Methods: Using the self-developed "Gui Women’s System" to establish a database of 1.69 million fetal cases in Guangxi and to analyze the distribution of time, space and population, as well as the outcomes of pregnancy, using the big data. Results: During the 29 months of observation, the overall live birth rate was 99.25%, with stillbirth rate during pregnancy as 0.44%, stillbirth rate during birth as 0.02%, and the 0-6 days mortality rate as 0.14%. The total detection rate on birth defects was 197.63/10 000; the incidence rate was 103.04/10 000, the birth rate was 102.55/10 000. The overall discovery rate of major birth defects was 48.33/10 000, with the incidence rate as 783 000, the birth rate as 0.58/10 000. The discovery rates of major birth defects in 14 cities were between 35 and 68/10 000, and the birth rate dropped significantly to less than 1.00 in 10 000. Nationalities showed that the number of pregnant women with birth defects more than 50 000 would include Hui (9.68/10 000), Yao (9.57/10 000), and Jing (9.37/10 000). With the increasing age of gestation, number of birth defects, incidence of major birth defects also increased. Ninety-five percent of the major birth defects were found within <28 weeks and with the top 5 kinds of major birth defects as complicated congenital heart disease (9.11/10 000), alpha thalassemia (8.36/10 000), and 21-trisomy syndrome (7.85/10 000), beta thalassemia (5.32/10 000) and fetal edema syndrome (4.92/10 000). The top 5 major birth defects appeared as complicated congenital heart disease (9.11/10 000), alpha thalassemia (8.36/10 000), and 21-trisomy syndrome (7.85/10 000), beta thalassemia (5.32/10 000) and fetal edema syndrome (4.92/10 000). Conclusion Programs leading to increase the rate on discovery of major birth defects were fundamental in effectively reducing the major birth defects.

OBJECTIVE: To evaluate the operability and effectiveness of a self-developed patellar bone canal locator (hereinafter referred to as "locator") in the reconstruction of the medial patellofemoral ligament (MPFL). METHODS: A total of 38 patients with recurrent patellar dislocation who met the selection criteria admitted between January 2022 and December 2022 were randomly divided into study group (the patellar canal was established with a locator during MPFL reconstruction) and control group (no locator was used in MPFL reconstruction), with 19 cases in each group. There was no significant difference in baseline data between the two groups ( P>0.05), such as gender, age, body mass index, disease duration, patella Wiberg classification, constituent ratio of cartilage injury, Caton index, tibia tubercle-trochlear groove, and preoperative Lysholm score, Kujal score, Tegner score, visual analogue scale (VAS) score, and so on. The Lysholm score, Kujal score, Tegner score, and VAS score were used to evaluate knee joint function before operation and at 3 days,1 month, 3 months, and 6 months after operation. The ideal prepatellar cortical thickness and canal length were measured before operation, and the actual prepatellar cortical thickness and canal length after operation were also measured, and D1 (the distance between the ideal entrance and the actual entrance), D2 (the ideal canal length minus the actual canal length), D3 (the ideal prepatellar cortical thickness minus the actual prepatellar cortical thickness) were calculated. RESULTS: Patients in both groups were followed up 6-8 months (mean, 6.7 months). The incision length and intraoperative blood loss in the study group were smaller than those in the control group, but the operation time was longer than that in the control group, the differences were significant ( P<0.05). There was no complication such as incision infection, effusion, and delayed healing in both groups, and no further dislocation occurred during follow-up. One patient in the study group had persistent pain in the anserine area after operation, and the symptoms were relieved after physiotherapy. The VAS score of the two groups increased significantly at 3 days after operation, and gradually decreased with the extension of time; the change trends of Lysholm score, Kujal score, and Tegner score were opposite to VAS score. Except that the Lysholm score and Kujal score of the study group were higher than those of the control group at 3 days after operation, and the VAS score of the study group was lower than that of the control group at 3 days and 1 month after operation, the differences were significant ( P<0.05), there was no significant difference in the scores between the two groups at other time points ( P>0.05). Patellar bone canal evaluation showed that there was no significant difference in preoperative simulated ideal canal length, prepatellar cortical thickness, and postoperative actual canal length between the two groups ( P>0.05). The postoperative actual prepatellar cortical thickness of the study group was significantly smaller than that of the control group ( P<0.05). D1 and D3 in the study group were significantly higher than those in control group ( P<0.05), but there was no significant difference in D2 between the two groups ( P>0.05). CONCLUSION The locator can improve the accuracy of MPFL reconstruction surgery, reduce the possibility of intraoperative damage to the articular surface of patella and postoperative patellar fractures.
Humans ; Patella/surgery* ; Patellar Dislocation/surgery* ; Patellofemoral Joint/surgery* ; Knee Joint/surgery* ; Joint Dislocations ; Ligaments, Articular/surgery*