Objective:Understanding of subset、memory cell generation、effector function and organ distribution of Th1 cells in vitro and in vivo.Methods:Different stages of polarized Th1 cells are generated from antigen-specific CD4 +T cells.The expression of cell surface markers and intracellular cytokines(IL-2、IFN-? and IL-4) is determined at a single cell level by FACS. Equal numbers of CFSF-labeled naive CD4 +T cells and different stages of Th1 cells are adoptively transferred into mouse.The production of IFN-? and distribution of Th1 memory cells in lymphoid and non-lymphoid organs are measured. Results:Th1 cell populations can be subdivided into several subsets based upon the production of IL-2 and IFN-?. The percentage of IFN-?-producing cells is increased and IL-2-producing cells is decreased following repeatedly stimulation under Th1 culture condition. Moreover,the expression of CD62L on Th1 cells is reduced after activation and differentiation of CD4 +T cells. 2 weeks post-transfer,the frequency of naive CD4 +T cells is comparable from lymph nodes、spleens and lungs,whereas memory Th1 cells are preferentially detected in spleens and lungs and rapidly produce IFN-? following re-exposure to the same antigen.Conclusion:Th1 cell population is composed of distinct subsets of cells. In comparson with naive CD4 +T cells,memory Th1 cells reveal the changes of surface molecule expression、biologic function and distribution in tissues.

Objective:To investigate the correlation between CD4+T cell division and surface molecule expression or cytokine production after stimulation with antigen.Methods:Isolation of CD4+T cells from spleens and lymph nodes of OVA-T cell-receptor transgenic mouse (TCR-Tg ). After stimulation with OVA peptide antigen in the presence of antigen-presenting cells, CD4+T cell division, surface molecule expression and intracellular cytokine production are determined in flow cytometry.Results:CD4+T cells are divided 1-5 times after stimulation for 3 days with antigen. The cell division is associated with both up- and down-regulation of surface molecules such as CD25, CD44, CD62L and CD69 and of cytokines such as IFN-?, IL-4 and IL-10.IL-12 promotes cell division and enhances IFN-? but inhibits IL-4 and IL-10 production.Conclusion:After stimulation with antigen, CD4+T cells are divided in association with the quantitative and qualitative changes of surface molecules and cytokine production.

Objective To evaluate the efficacy and toxicity of Taxol-based chemotherapy in patients with advanced gastric cancer. Methods Thirty-one patients with advanced gastric cancer were treated with two kinds of Taxol-based chemotherapy schedule. The treatment schedule were composed of paclitaxel and fluorouracil(TF) or cisplatin(TP). The patients who had had chemotherapy with fluorouracil were arranged to TP group,others to TF group. A treatment cycle lasted 21 days. The efficacy was evaluated after two cycles of the chemotherapy. Results total efficiency rate of two groups was 45.2 %; the efficiency rate of TF was 47.4 %; the rate of TP was 41.7 %; CBR was 80.6 %. The main side effect of toxicity were neutropenia and nausea as well as vomiting. Conclusion Taxol-based combination chemotherapy is a high response protocol with mild toxicity. TP schedule has a high efficiency for advanced gastric cancer.

Objective To investigate the correlation between quantitative parameters with contrastenhanced ultrasound (CEUS) and microvessel density (MVD).Methods Thirty-four placenta of rat model of preeclampsia underwent CEUS examination.The peak intensity time curves on the enhanced images were analyzed quantitatively with computer to get quantitative parameters[the time to peak(TTP), maximal peak intensity(Imax), the area under curve(AUC) and the mean perfusion volume(V)].These parameters were compared with MVD counted with immunohistochemistry and the correlation was statistically studied.Results The TTP in the enhanced images was (14.55 ± 3.45)s, Imax was (20.83 ± 6.15) dB, AUC was (1868.61 ± 25.76)dB, V was (58.01 ± 23.56)dB, and the MVD of placenta of rat model of preeclampsia was (88.98 ± 24.78) in 34 rats.The Imax was correlated positively to MVD (r = 0.885, P = 0.000) ,AUC was correlated positively to MVD (r = 0.677, P = 0.001), V was correlated positively to MVD (r =0.877, P = 0.000).There was no correlation between TTP and MVD in lesions.Conclusions The Imax,AUC and V calculated with CEUS were correlated to MVD, these parameters were valuable index for quantitative evaluation of placental blood perfusion.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Combined Modality Therapy ; External Fixators ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Radius Fractures ; physiopathology ; therapy ; Splints

Objective:To investigate the effect of intercellular adhesion molecule-1 (ICAM1) targeted miR-133a-5p on lipopolysaccharide (LPS)-induced alveolar epithelial A549 cell injury.Methods:Dual lucife-rase reporter assay was used to verify the ICAM1 targeted effect of miR-133a-5p.A549 cells were induced by LPS in vitro and divided into the control group, LPS group, LPS+ negative control(miR-NC) group, LPS+ miR-133a-5p group, LPS+ small interfering RNA(si)-NC group, and LPS+ si-ICAM1 group.The expression levels of miR-133a-5p and ICAM1 mRNA were detected by real-time fluorescent quantitative PCR.Apoptosis was detected by flow cytometry.The expression levels of ICAM1, Bcl-2, Bax and cleaved caspase-3 protein that could activate cysteine were detected by Western blot.The expression levels of interleukin-6 (IL-6) and tumor necrosis factor α(TNF-α) were detected by enzyme linked immunosorbent assay kit. Results:Compared with the control group, the LPS group had a decreased expression level of miR-133a-5p (0.39±0.04 vs.1.00±0.09) in A549 cells, increased expression of ICAM1 (0.86±0.08 vs.0.39±0.03), an increased apoptotic rate [(27.65±2.47)% vs.(8.13±0.89)%], and increased secretion of IL-6 [(624.59 ± 51.42) ng/L vs.(194.25±18.43) ng/L] and TNF-α [(548.35±51.42) ng/L vs.(174.26±19.43) ng/L]. The differences were significant (all P<0.05). Compared with the LPS+ miR-NC group, the apoptosis rate of A549 cells [(13.46±1.38)% vs.(28.71±2.54)%] in LPS+ miR-133a-5p group were significantly decreased, and the secretion of IL-6 [(296.43±23.51) ng/L vs.(635.86±55.41) ng/L] and TNF-α [(321.14±30.56) ng/L vs.(563.24±49.52) ng/L] was significantly decreased (all P<0.05). Compared with LPS+ si-NC group, the apoptosis rate of A549 cells [(13.65±1.64)% vs.(23.51±2.33)%] in LPS+ si-ICAM1 group was significantly decreased, and the secretion of IL-6 [(324.15±29.41) ng/L vs.(625.39±52.59) ng/L] and TNF-α [(334.65±20.46) ng/L vs.(534.97±51.42) ng/L] were significantly decreased (all P<0.05). Conclusions:miR-133a-5p can alleviate LPS-induced alveolar epithelial cell injury, and the mechanism may be related to down-regulation of ICAM1 expression.

Based on the theory of the uterus is an extraordinary fu （腑） organ， it is believed that the core pathogenesis of precocious puberty in girls is the dysfunction of the uterus in storing and draining， while the basis of the disease is the delicate and vulnerable zang fu （脏腑） organs； the imbalance of qi and blood in the Chong and Ren mai （冲任） is the key link of the pathogenesis， and five zang （脏） organs dysharmony is responsible for the prolongation of the disease. For treatment， it is advocated to conform to the physiological characteristics that zang fu organs are delicate and fragile in girls， focus on regulating the uterus， and take into account the harmonization of the zang fu organs； the method of consolidating and nourishing yin is suggested as the basic treatment method， and self-made Xingzaoshu Formula （性早熟方） is commonly used. Considering that the uterus of girls is delicate and vulnerable， neutral medicinals are mainly used， and drastic supplementation and strong attack are avoided； pulse administration of Chinese herbal medicine is advocated. Since the condition of precocious puberty is often prolonged， Chinese herbal medicine together with medicinal diet is often used in the later stage of treatment to harmonize the zang fu organs， and self-made medicinal diet Xingzaoshu Soup （性早熟煲） is often used.

Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells to provide an experimental basis for cartilage repair.Methods Bone marrow mesenchymal stem cells(BMSCs)were cultured using the Percoll density gradient separation method,and this was followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints and subjected to physical crushing,repeated freeze-thaw cycles,and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of the decellularized scaffolds at different concentrations,three groups of scaffolds(labelwd A,B,and C)were designed with concentrations of 100％,50％and 30％,with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto optimally concentrated scaffolds and cultured for 1 week to observe cell growth.Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45.Osteogenic induction stained with alizarin red showed red calcific nodules,and chondrogenic induction stained with alcian blue showed blue cartilaginous nodules.No apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin-eosin,and toluidine blue.There was a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds(P＜0.05).The content of glycosaminoglycans was slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size,water absorption,porosity,tensile strength,and Young's modulus(P＜0.05).After co-cultivation of stem cells with the scaffolds,cell adhesion was found to be good.Conclusions Percoll density gradient separation can obtain high-purity rabbit BMSCs,and the mixed decellularization method is superior.Group B scaffolds were the most suitable for tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group B scaffolds.

[Abstract] Objective: To investigate the effect of Tim-4 on invasion and migration of cervical cancer cells and its underlying mechanisms. Methods：The expression levels of Tim-4 in cervical cancer cell lines Siha, Hela and cervical epithelial immortalized cell line H8 were detected by Real-time PCR. The Tim-4 lentiviral vector was transfected into Siha cell line. The over-expression of Tim-4 in Siha cell line was detected by fluorescence microscopy. The effects of Tim-4 on the invasion and migration of cervical cancer cell line were detected by Transwell and scratches methods. The changes of MMP2, MMP9, E-cadherin and N-cadherin in Siha cells were detected by Western blotting. Results：The expression of Tim-4 was higher in Siha and Hela cell lines compared to that in the H8 cell line. The Siha cell line burdening Tim-4 lentiviral vector was successfully constructed. Over-expression of Tim-4 significantly inhibited the migration and invasion of cervical cancer cell line, and affected the expression of MMP2, MMP9, N-cadherin and E-cadherin. Conclusion：Over-expression of Tim-4 promotes the invasion and migration by regulating the EMT transformation in cervical cell carcinoma.