OBJECTIVE To investigate the effects of galangin （GLA） on autophagy and apoptosis of chondrocytes in knee osteoarthritis （KOA） rats by regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/UNC-51-like kinase 1 （ULK1） signaling pathway. METHODS KOA rat model was constructed and separated into model group， L-GLA， M-GLA， H-GLA groups [subcutaneous injection of 100， 200， 400 μg/kg GLA]， GLA+Compound C group [subcutaneous injection of 400 μg/kg GLA+0.2 mg/kg AMPK inhibitor Compound C]， with 10 rats in each group. Additionally， 10 normally fed rats were selected as the sham operation group. After the last medication， the degree of knee joint swelling of rats in each group was detected； the pathology of knee joints in KOA rats was observed. The serum expressions of matrix metalloproteinase 13 （MMP-13） and interleukin-1β （IL-1β） in KOA rats were detected； the autophagy of chondrocytes in KOA rats was observed； the chondrocyte apoptosis in KOA rats was detected； the phosphorylation of AMPK/mTOR/ULK1 pathway-related proteins in cartilage tissue of knee joint were detected in rats. RESULTS Compared with the sham operation group， the arrangement of articular chondrocytes in the model group was disordered， with nuclear pyknosis and severe fibrosis of the articular cartilage layer， accompanied by a large amount of inflammatory cell infiltration； the degree of joint swelling， the number of autophagic vacuoles and apoptosis rate of chondrocytes， serum levels of MMP-13 and IL-1β， and the phosphorylation of mTOR protein in cartilage tissue of knee joint were all increased significantly （P＜0.05）， while the phosphorylation of AMPK and ULK1 protein were all decreased significantly in cartilage tissue of knee joint （P＜0.05）. Compared with the model group， L- GLA， M-GLA， H-GLA groups showed significant improvement in joint cartilage injury and reduced infiltration of inflammatory cells in rats. The above quantitative indicators were significantly reversed in a dose-dependent manner，except the number of autophagic vacuoles increased significantly （P＜0.05）. Compared with the H-GLA group， the GLA+ Compound C group showed aggravated cartilage tissue of joint cartilage injury and inflammatory cell infiltration in rats， and the above quantitative indicators were reversed significantly （P＜0.05）. CONCLUSIONS GLA can promote autophagy and inhibit apoptosis of chondrocytes in KOA rats， the mechanism of which may be associated with activating AMPK/mTOR/ULK1 signaling pathway.

Objective:To analyze the changes in biological characteristics including infectivity, growth and pathogenicity of Chlamydia muridarum ( Cm) after serial passage in vitro in special conditions in order to provide reference for screening attenuated live vaccines and virulence-related genes. Methods:Wild-type Cm strain (G0) was cultured for several passages using conventional cell culture method under alternate unassisted and assisted culture conditions. Then, the 28th generation (G28) of Cm was selected and compared with the parental G0 strain in terms of centrifugation dependence, attaching ability, intracellular growth curve, plaque size and fallopian tube lesions after genital tract infection in a mouse model. Results:Compared with the parental G0 strain, the G28 strain showed significantly decreased dependence on centrifugation during cell infection ( P<0.05) and increased attachment capacity to cells ( P<0.05). No significant differences were observed in the growth curves 32 h after cell infection or in the plaque sizes between the parental G0 and G28 strains. In the in vivo virulence test, fallopian tube lesions were observed in 87.5% of G0-infected mice and 37.5% of G28-infected mice ( P<0.05). Conclusions:Compared with the parental G0 strain, the G28 strain showed significantly enhanced in vitro infection ability, but decreased in vivo pathogenicity, which brought hope for further identification of virulence genes, isolation of attenuated strains with single genotype and development of live attenuated Chlamydia vaccines.

First-class education for postgraduate is the foundation for the construction of "double first-class" education. Universities play an important role in postgraduate education. This paper explored the measures for the reform and innovation of the construction of "double first-class" education for postgraduate students in our university, which include perfecting the supervisor's responsibility and authority mechanism, deepening the reform on curriculum system, strengthening the construction of sharing platform, and improving the evaluation mechanism of training quality and so on. In conclusion, initial achievement from the reform and innovation of training mode was observed, which provides a useful reference for the construction of "double first-class" education for local universities.

OBJECTIVETo analyze the core acupoints and compatibility of electroacupuncture (EA) for simple obesity based on complex network technique, and to explore the usage of EA waveform.

METHODSThe clinical research literature regarding EA for simple obesity published from January of 1980 to June of 2016 were searched in PubMed, CNKI, , VIP, CBM and TCM online database to establish a prescription database of EA for simple obesity. The Matlab2014a software was used to perform the center analysis and cluster analysis, and the analysis of core points and compatibility were conducted. Gephi 9.1 software was used to demonstrate the complex network diagram to further analyze the usage of EA waveform.

RESULTSTotally 238 prescriptions were obtained. The selection of acupoints at -meridians were equally important with acupoints at -meridians. The meridians with highest core degree were stomach meridian, conception vessel and spleen meridian. The acupoints with highest core degree were Sanyinjiao (SP 6), Tianshu (ST 25) and Zusanli (ST 36). The cluster analysis indicated three acupoint clusters, including the key-acupoint cluster, syndrome-acupoint cluster, and -point cluster; it was revealed Tianshu (ST 25) and Zhongwan (CV 12) had the highest intensity of compatibility. The sparse-dense wave was mostly used in EA for simple obesity, followed by continuous wave, indicating both sparse-dense wave and continuous wave had high clinical application value.

CONCLUSIONThe acupoints of EA for simple obesity are mainly in stomach meridian, conception vessel and spleen meridian; sparse-dense wave is mostly used, followed by continuous wave.

Objective To analyze the protein expression profile of HeLa cells transfected with pORF5 gene of Chlamydia trachomatis. Methods A lentiviral expression vector containing pORF5 gene was constructed. The lentiviral expression vector and helper plasmids were co-transfected into 293T cells to construct the recombinant lentivirus, which was used to infect HeLa cells. HeLa cells transfected with pORF5 gene and control HeLa cells were sorted out by flow cytometry. The isobaric tags for relative and absolute quantitation ( iTRAQ) approach combined with nano-liquid chromatography-tandem mass spec-trometry ( NanoLC-MS/MS) analysis was performed to understand protein expression profiles and to iden-tify and quantify the differentially expressed proteins in the pORF5-transfected HeLa cells ( pORF5-Hela) and the control HeLa cells. Quantitative real-time PCR ( qRT-PCR ) and Western blot analysis were performed to detect the expression of some proteins at mRNA and protein levels, respectively. Results HeLa cell line stably transfected with pORF5 gene and control HeLa cell line were constructed successful-ly. Totally 314 proteins were differentially expressed between the pORF5-HeLa and control HeLa cells, 159 of which showed increased expression and the other 155 showed decreased expression in pORF5-HeLa cells. The differentially expressed proteins were involved in many processes, such as metabolic process, immune response, biological adhesion and so on. Results of qRT-PCR showed that the expression of HIST1H1C(histone H1. 2C), HBA1(hemoglobin subunit alpha), PARK7(parkinson disease protein 7), HMGB1(high mobility group protein B1) and HMGB2 at mRNA level in pORF5-HeLa cells were up-regulated, while the expression of CLIC1 ( chloride intracellular channel protein 1 ) , KRT7 ( typeⅡ cy-toskeletal 7), SFN(14-3-3 protein sigma) and CDKN2A(cyclin-dependent kinase inhibitor 2A) were down-regulated. Western blot analysis confirmed the enhanced expression of HMGB1 and PRAK7 at pro-tein level. The results of qRT-PCR and Western blot analysis were consistent with proteomic data. Con-clusion Expression profiles for differentially expressed proteins between pORF5-HeLa and control HeLa cells were established successfully. The differentially expressed proteins regulated by pORF5 gene were found to be related to cell metabolism, proliferation, adhesion and so on, suggesting that pORF5 might promote the growth and proliferation of Ct by regulating protein expression and biological behavior of host cells.

Objective To observe the clinical efficacy of acupoint thread embedding on the peripheral serum leptin and insulin levels in simple obesity patients, and to further explore the mechanism of acupoint thread embedding in treating simple obesity.Method A total of 120 patients with simple obesity were randomized into a control group, an eletroacupuncture (EA) group and a thread embedding group, 40 cases in each group. The control group was intervened by diet plus exercise intervention, the EA group by EA treatment in addition to the intervention given to the control group and the thread embedding group by acupoint thread embedding treatment in addition to the intervention given to the control group. The levels of blood serum leptin and insulin in the three groups were observed before and after 2 treatment courses and the clinical efficacies were compared among the three groups.Result The total effective rate was 87.5% in the thread embedding group, 85.0% in the EA group, and 47.5% in the control group. The total effective rates in the thread embedding group and EA group were significantly different from the rate in the control group (P<0.05). The levels of the fasting serum leptin and insulin were significantly changed in the thread embedding group and EA group after the treatment (P<0.01). After the treatment, the levels of the fasting serum leptin and insulin in the thread embedding group and EA group were significantly different from those in the control group (P<0.01,P<0.05). The insulin level in the thread embedding group was better than that in the EA group, and the difference was statistically significant (P<0.05).Conclusion Acupoint thread embedding is an effective approach for simple obesity, and it can down-regulate the fasting peripheral serum leptin and insulin levels.

Objective To evaluate inhibitory effect of Chlamydia trachomatis plasmid-encoded protein pORF5 on HeLa cell apoptosis induced by tumor necrosis factor-alpha(TNF-α). Methods The recombinant lentiviral expression vector containing pORF5 gene and helper plasmids were co-transfected into 293T cells to prepare the recombinant lentivirus. Then, the lentivirus particles were collected and concentrated, and used to infect HeLa cells. Flow cytometric screening identified stable pORF5-expressing HeLa (pORF5-HeLa) cells. Meanwhile, the empty plasmid was transfected into HeLa cells to prepare control HeLa cells. The two cell lines were both divided into two subgroups to be treated with 20μg/L TNF-αand fresh culture medium respectively for 6 hours. Then, Hoechst 33258 staining was performed to observe morphological changes of apoptotic cells, flow cytometry to detect cell apoptosis, real-time PCR to measure the mRNA expression of Caspase3, Bax and Bcl-2, and Western blot analysis to determine the protein expression of Bax and Bcl-2. Results After 6-hour treatment with TNF-α, Hoechst 33258 staining showed variable degrees of karyopyknosis and karyorrhexis, and highly-refractive blue apoptotic bodies in the pORF5-HeLa cells and control HeLa cells. The pORF5-HeLa cells and control HeLa cells both showed significantly higher apoptosis rate in the treated subgroup than in the untreated subgroup (pORF5-HeLa cells:35.5%± 4.5%vs. 9.5%± 1.5%, t=13.53, P<0.01;control HeLa cells:63.6%± 5.8%vs. 7.9%± 0.9%, t=32.36, P<0.01). Compared with treated control HeLa cells, treated pORF5-HeLa cells showed significant decreases in mRNA expression of Bax(72.8%)and Caspase 3(84.5%)(t = 35.29, 42.25, respectively, both P < 0.01), as well as in Bax protein expression(t = 17.58,P < 0.01), but significant increases in Bcl-2 mRNA and protein(6.8 times)expression(t = 87.12, 18.93, respectively, both P <0.01). Conclusion pORF5 plasmid protein can inhibit TNF-α-induced HeLa cell apoptosis likely by increasing the expression of anti-apoptotic protein bcl-2 and decreasing the expression of pro-apoptotic proteins Caspase-3 and Bax.

Objective:To observe the changes of metabolic intensity value of points on the face before and after needling bilateral Hegu (LI 4) in healthy people and provide scientific basis for association between Hegu (LI 4) and face/mouth. Methods:A total of 45 healthy college students were selected in this study. Using medical thermography and Pennes bio-heat transfer model, the infrared thermograph images on the face before and after needling bilateral Hegu (LI 4) were collected to observe the distribution of metabolic intensity value on the face before acupuncture and changes in these values after needling bilateral Hegu (LI 4). Results:Before acupuncture, Cuanzhu (BL 2) had the maximal metabolic intensity value. Its mean value was (0.71±0.23) W. Quanliao (SI 18) had the minimal metabolic intensity value. There were no left-right statistical significances in metabolic intensity values. After needling bilateral Hegu (LI 4), the metabolic intensity values of most points on the face were increased. Kouheliao (LI 19) obtained the maximal increase: 0.35 W on average; and Yangbai (GB 14) obtained the minimal increase: 0.08 W on average. Conclusion:Points on both sides in healthy people have good symmetry in metabolic intensity value. After needling bilateral Hegu (LI 4), the metabolic intensity values of points on the face were increased, especially points around the lips, which accords with the pathway of the Large Intestine Meridian on the head and face. This provided some scientific foundation for the association between Hegu (LI 4) and face/mouth.

OBJECTIVE:To explore risk factors for the death in HIV/AIDS patients suffering from myelosuppression during highly active antiretroviral therapy(HAART)treatment. METHODS:The historical cohort study method was used to choose 735 in-patients from Nanning Forth People's Hospital during Jan. 2011 to Jul. 2015. Univariate and multivariate Logistic regression analy-sis were used to show the risk factors for the death. RESULTS:Of 735 cases,there were 648 survival cases and 87 dead cases, with mortality of 11.8%. Univariate Logistic analysis showed that:male,elder,higher total bilirubin,lower creatinine clearance, lower CD4+T cell count in baseline,combined more opportunistic infection,combined more myelosuppressive drugs,thrombocyto-penia,lower hemoglobin were the risk factors for the death of HIV patients with myelosuppression in HARRT treatment,while route of HIV infection,HAART treatment A including Zidovudine,weight were the protective factors. Multivariate Logistic regres-sion analysis showed male,elder,higher total bilirubin,lower creatinine clearance,lower CD4+ T cell count in baseline,com-bined more opportunistic infection,combined more myelosuppressive drugs,thrombocytopenia,lower hemoglobin were the risk factors for the death of HIV patients with myelosuppression in HARRT treatment too. CONCLUSIONS:Pertinence treatment and control methods for HIV/AIDS with myelosuppression in HARRT treatment should be taken to reduce the mortality in the future.

Objective:To investigate whether pORF5 plasmid protein of Chlamydia trachomatis(Ct) induces 1L-1βand 1L-18 production in THP-1 cells,and its potential molecular mechanism.Methods:pORF5 plasmid protein was used to stimulate THP-1 cells at different concentrations(0,3,6,12,24,36 μg/ml),then the inflammatory cytokines IL-18 and IL-1βwere detected by ELISA at the time of 0,8,16,24,36 h;The mRNA expression of NALP3 inflammasome were detected by Realtime-PCR,and Caspase-1 activity was determined by Western blot analysis.THP-1 cells were transfected with siRNA targeting NALP3 and ASC gene for 24 h or pretreated with Caspase-1 inhibitor(Z-YVAD-FMK) for 30 min,and subsequently stimulated with pORF5(24 μg/ml) for 24 h,then secretion of IL-1βand IL-18 were analyzed by ELISA.Results: The pORF5 plasmid protein induced THP-1 cells to secrete IL-1βand IL-18 by dose-and time-dependent manners,production of IL-1βand IL-18 reached their peaks(491 pg/ml and 186 pg/ml) at concentration of 24 μg/ml,and the peak amount of IL-1βand IL-18 occurred at 24 h and 16 h post-stimulation respectively.pORF5 plasmid protein in-creased mRNA expression of NALP3 inflammasome and activated Caspase-1 in THP-1 cells.NALP3 siRNA,ASC siRNA and Z-YVAD-FMK reduced pORF5-induced IL-1βand IL-18 production when compared with control groups(P<0.05).Conclusion:pORF5 plasmid protein could induce THP-1 cells to produce IL-1βand IL-18 through NALP3 inflammasome activation,which may play an important role in the pathogenesis in Ct infection.