Objective To investigate the prevalence of resistant Neisseria gon orrhoeae and plasmid-mediated resistant strains in Guangzhou from 1996 to 2001. Methods The resistant N.gonorrhoeae and plasmid-mediated resistant strains to tetracycline (TRNG) were determined using agar dilution method, and penicillinas e-producing N. gonorrhoeae (PPNG) by acidometric method. Results A total of 793 gonococcal isolates were tested from 1996 to 2001. The resistant rate for penic illin increased from 57.2%to 81.8%and PPNG from 2%to 27.2%, respectively, du ring the six years. Resistance to tetracycline remained high and stable over the years, while the rates of TRNG were increased from 1.5%to 27.2%. Conclusions The prevalence of plasmid-mediated resistant strains of N. gonorrhoeae increase s year by year in Guangzhou. These results suggest that the clinical isolates of gonococcal strains in this city are highly resistant to penicillin and tetracyc line.

Objective To investigate the feasibility and prospects of nested real-time PCR(NR-PCR)technique for Treponema palladium(Tp)detection in various samples of different stages of syphilis from patients preliminarily diagnosed as syphilis. Methods Targeting the Tp polA gene, NR-PCR was performed to detect Tp DNA in various samples from the patients with various stages of syphilis at the first clinic visit, including skin tissue fluid swabs, serum, whole blood, cerebrospinal fluid(CSF)and earlobe blood. Data were analyzed with SPSS software version 13. Results A total of 368 clinical samples were collected from 200 patients with syphilis. With a detection limit of 2 Tp/ml, NR-PCR showed that the total positive rate for Tp DNA was 71.7%(264/368). The Tp DNA positive rate was highest in earlobe blood samples (92.0%, 23/25), followed by CSF samples(90.2%, 46/51), skin tissue fluid swabs(74.3%, 26/35), serum samples(66.9%, 99/148)and whole blood samples(64.2%, 70/109). There was good agreement between NR-PCR results and serologic test results, with a consistency rate of 76.0%(152/200). Furthermore, the Tp DNA positive rate did not differ between patients with primary(12/19)and secondary syphilis(14/16)in skin tissue fluid swabs(χ2 = 2.62, P > 0.05), and was slightly but insignificantly higher in patients with secondary syphilis than those with primary syphilis in the serum samples(χ2=3.6, P=0.06). The Tp DNA positive rate of whole blood samples was also higher in patients with secondary syphilis than those with any other types of syphilis. Among patients with neurosyphilis, no significant difference was observed in the Tp DNA positive rate between earlobe blood samples and CSF samples(P=0.06). Among patients with latent syphilis, the Tp DNA positive rate was significantly higher in serum samples with an RPR titer of ≥ 1:8 than those with an RPR titer of≤1:4. Conclusion NR-PCR is feasible for detecting Tp DNA in various kinds of samples, and the Tp DNA positive rate is influenced by stages of syphilis and types of samples, as well as RPR titers.

Objective The present study is to investigate IL-24 gene(Ad5F35-hIL-24) effect on the topoisommeraseⅡα(topoⅡα) and Caspase-3 expression in glioma cell line U251. Methods After transfected the U251 glioma cells with the Ad5F35-hIL-24, the methyl thiazolyl tetrazolium (MTT) was used to analyse the inhibition rate of Ad5F35-hIL-24 on the cells. Hoechst 33258 fluorescent staining and flow cytometric assay were used to detect apoptosis. The immunohistochemistry assay was used to detect topoⅡα expression, and Western blotting was applied to detect the protein expression of topoⅡα and caspase-3. Transwell experiment was used to test the invasiveness of the cells. Results It was found that the Ad5F35-hIL-24 could inhibit U251 cell proliferation and induce apoptosis in a dose dependent manner compared with the control groups. It showed that Ad5F35-hIL-24 could inhibit topoⅡα expression reveled by immunohistochemistry and Westeren blotting, while it increased caspase-3 protein expression. The Transwell experiment showed that the Ad5F35-hIL-24 could reduce the invasiveness of the U251 glioma cells.Conclusion The exogenous IL-24 gene can inhibit the cell proliferation and induce apoptosis of U251 glioma cells. The topoⅡα and Caspase-3 are the important molecular targets of the IL-24 gene. These results may give support for the IL-24 gene usage in clinical treatment for glioma patients.

Objective To analyze the characteristics of genotyping and gene polymorphism of Neisseria gonorrhoeae(N.go) with azithromycin(AZM)-resistance(AZM-R) and decreased susceptibility to ceftriaxone(CROD).Methods The minimum inhibitory concentration(MIC) of AZM and CRO were determined.AZM-R isolates were detected for mutations in 23S rRNA,mtrR and penA genes.Genotypes were analyzed by using N.go multi-antigen sequence typing(NG-MAST).Results All total of 485 isolates of N.go were detected.77(15.9%) strains were AZM-R(MIC≥1 mg/L),including 33(6.8%) isolates of AZM low-level resistant(AZM-LLR,MIC=1 mg/L) strains and 44(9.1%) isolates of AZM middle-level resistant(AZM-MLR,MIC≥2 mg/L) strains.There were more CROD(MIC≥0.125 mg/L) strains in AZM-MLR isolates(43.2%),compared with those in AZM-LLR isolates(18.2%,P<0.05).The detected rates of 23S rRNA,mtrR,penA single or combined mutations were without significant differences between AZM-LLR isolates and AZM-MLR isolates(P>0.05).Similar results were found between combined AZM-LLR/CROD isolates and combined AZM-MLR/CROD isolates(P>0.05).No mutation of A2059G and AZM high-level resistant(AZM-HLR,MIC≥256 mg/L) isolate were found.Among 77 AZM-R isolates,67 sequence types(ST) were identified by NG-MAST,of which 30 types were novel.Most ST were represented by a single isolate.Conclusion AZM-R and CROD isolates,presented in this area,might be deserved continuous surveillance to identify the mechanism of concurrent resistance.

Objective To investigate genetic characteristics of Neisseria gonorrhoeae (N.gonorrhoeae) isolates from Guangzhou city in 2014,and to analyze the relationship of N.gonorrhoeae multi-antigen sequence typing (NGMAST) sequence types (STs) with ciprofloxacin resistance.Methods An agar dilution method was used to determine the minimal inhibitory concentration (MIC) of ciprofloxacin in 97 N.gonorrhoeae isolates from Guangzhou city.PCR was performed to amplify the gyrA,parC,porB and tbpB genes from these isolates,followed by gene sequencing and determination of NG-MAST STs.Results Of the 97 N.gonorrhoeae isolates,95 (97.9％) were resistant to ciprofloxacin,including 41 high-level (MIC ≥ 16 mg/L) and 54 low-level (1 mg/L ≤ MIC ＜ 16 mg/L) resistant strains.Mutations were detected at codons 91 and 95 encoding serine in the gyrA gene of all the 95 ciprofloxacin-resistant strains,and in the parC gene of 93 resistant strains.The frequency of the mutation at codon 87 in the parC gene was 85.4％ (35/41) in high-level resistant strains,significantly higher than that in low-level resistant strains (59.3％[32/54],x2 =7.64,P ＜ 0.05).MAST STs were successfully determined for all the 97 N.gonorrhoeae isolates except 1 isolate with incorrect PCR amplicons.Of the 96 genotyped isolates,50 were assigned to 35 known STs by using the NG-MAST website (www.ngmast.net),among which,10 STs each contained 2 to 4 isolates.The most common ST was ST5309.Phylogenetic tree analysis revealed that the 96 genotyped N.gonorrhoeae isolates could be classified into 2 groups,and the proportion of isolates with MIC ≥ 16 mg/L is 46.4％ (39/84) in group 1,but only 1/12 in group 2 (x2 =6.27,P=0.012).Conclusions High-level resistance of N.gonorrhoeae to ciprofloxacin may be mainly associated with the mutation at codon 87 in the parC gene.NG-MAST STs may be related to the degree of ciprofloxacin resistance.

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Objective To compare the sensitivity and specificity of venereal disease research laboratory (VDRL) test versus several other laboratory tests in the diagnosis of neurosyphilis. Methods Lumber puncture was conducted to obtain cerebrospinal fluid (CSF) from untreated outpatients with latent syphilis (LS) or serofast outpatients with LS. Then, VDRL test, rapid plasma regain (RPR) test, Treponema pallidum particle agglutination (TPPA) assay, fluorescent treponemal antibody-absorption (FTA-ABS) test and protein quantification were performed on these CSF samples. The sensitivity, specificity, positive predictive value and negative predictive value were compared between VDRL test and four other laboratory tests in the diagnosis of neurosyphilis. Results Totally, 61 cases of latent syphilis were included in this study. The sensitivity, specificity,positive predictive value and negative predictive value were 93.44% (57/61), 99.32%(293/295), 96.61%(57/59), 98.65% (293/297)for CSF-RPR, respectively, 91.80% (56/61), 82.71% (244/295), 52.34% (56/107),97.99 (244/249) for CSF-TPPA, respectively, 93.44% (57/61), 82.71% (244/295), 52.78%(57/108), 98.39%(244/248) for CSF-FTA-ABS, respectively, and 49.18%(30/61), 97.29% (287/295), 78.95% (30/38),90.25% (287/318) for CSF protein quantification, respectively. Conclusions CSF-VDRL cannot be replaced by CSF-RPR, -TPPA, -FTA-ABS, or CSF protein quantification in the diagnosis of neurosyphilis. CSF-RPR shows a high sensitivity and specificity in the diagnosis of neurosyphilis, with an increased diagnostic capability (area under the receiver operating characteristic curve) compared with CSF-TPPA, CSF-FTA-ABS or CSF protein quantification.

Objective To evaluate the curative effect of Huoxue-Xiaozhi capsule on non-alcoholic fatty liver disease (NAFLD). Methods A total of 100 patients with NAFLD who met the inclusion criteria were divided into 2 groups with 50 patients in each group by random number table method. The treatment group was treated with Huoxue-Xiaozhi capsule, while the control group was treated with Compound Methionine Choline Tablets. Both groups were treated for 8 weeks. The ALT, AST, γ-GT (γ- glutamyltranspeptidase ), ALP (alkaline phosphatase, alkaline phosphate) were detected by automatic blood biochemical analyzer. The serum high sensitivity-C reactive protein (hs-CRP) and Homocysteine (Hcy) were detected by supplementary detection method. The serum IL-18 was detected by ELISA method, and liver imaging was performed before and after treatment. The symptoms and signs were evaluated, and clinical efficacy was evaluated. Results The total effective rate was 90.0％ (45/50) in the treatment group and 74.0％ (37/50) in the control group. The difference between the two groups was statistically significant (Z=-2.328, P=0.020 ). After treatment, the serum hs-CRP (2.4 ± 2.9 mg/L vs. 3.6 ± 2.8 mg/L, t=2.105), Hcy (11.2 ± 5.5 μmol/L vs. 13.9 ± 6.4 μmol/L, t=2.262), IL-18 (690.6 ± 61.3 ng/L vs. 775.4 ± 60.5 ng/L, t=6.962) in the treatment group were significantly lower than those in the control group (P<0.05 or P<0.01). After treatment, the improvement of hypochondriac lump, sallow complexion, dark complexion and hypochondriac dull pain in the treatment group were superior to those in the control group (Z values were -2.563, -2.788, -2.780, -2.220 respectively, all Ps<0.05 ). The Serum ALT, AST,γ-GT and ALP levels were significantly lower than those of the control group (t values were 18.820, 19.811, 10.221 and 3.248 respectively, all Ps<0.001 ). Conclusions The Huoxue-Xiaozhi capsule can improve the liver function and reduce the levels of serum hs-CRP, HCY and IL-18, and its curative effect of NAFLD.

Objective To observe the clinical therapeutic effect of Xiaozhong Sanjie prescription combined with Huayu powder external application for treatment of patients with nodular goiter (NG) and approach its mechanism. Methods A prospective randomized controlled study was conducted, including 128 patients who were definitely diagnosed as NG and admitted into Affiliated Hospital of Qinghai University from September 2014 to December 2016, and they were divided into a control group and a traditional Chinese medicine (TCM) treatment group by random number table method, 64 cases in each group. The control group received follow up observation; the TCM group was treated by Xiaozhong Sanjie prescription (the ingredients: rhizoma sparganii 15 g, zedoary 10 g, fritillary bulb 15 g, prunella vulgaris 20 g, bupleurum 6 g, dried tangerine 10 g, oyster 20 g, fructus aurantii 10 g, radix glycyrrhizae 5 g) in the mean time combined with Huayu powder (the ingredients: borneol 10 g, prunella vulgaris 20 g, pinellia ternata 15 g, oyster 15 g, radix curcumae 15 g, rhubarb 15 g); the ingredients of the prescription were immersed in water and decocted to form a decoction, a dose 400 mL daily, 200 mL taken orally in the morning and 200 mL in the evening after meal; the powder ingredients were mixed and stirred thoroughly with Chinese rice wine or vinegar (10 mL) to form a paste which was put uniformly flat onto a gauze about 15 cm in length and 1 cm in thickness, roasted by infrared lamp, as the paste temperature was lowered to body temperature, it was attached on the goiter skin of neck, then fixed by bandage before sleep to the next morning, afterwards the paste was removed and nodular skin washed, the length of external application being guaranteed to be over 4 hours a day; in the event when skin red swelling or pruritus occurred, promethazine should be used or the duration of application was shortened. Fifteen days constituted one therapeutic course, and after consecutive 3 courses of treatment, the therapeutic effects in the two groups were observed. Before and after treatment in the two groups, the changes of thyroid nodular size and TCM syndrome score were observed, meanwhile the thyroid functional index levels of serum vascular endothelial growth factor (VEGF), insulin-like growth factor Ⅰ(IGF-Ⅰ) and transforming growth factor-β1 (TGF-β1) were detected by enzyme-linked immunosorbent assay (ELISA). Results After treatment for 45 days, compared with control group, the thyroid nodule in TCM treatment group was significantly shrunken (mm: 8.75±3.41 vs. 15.89±4.51, P < 0.05) , and TCM syndrome score in TCM treatment group was significantly lowered (20.35±4.83 vs. 35.53±6.71, P < 0.05); before and after admission in the two groups, the thyroid function indexes of thyrotropic-stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4) were in the normal range, and there were no statistical significant differences between the two groups (all P > 0.05); the levels of serum VEGF, IGF-I were reduced in the TCM treatment group [VEGF (ng/L): 27.52±8.35 vs. 33.08±9.57, IGF-Ⅰ (μg/L): 24.12±6.74 vs. 35.57±11.39], TGF-β1 was increased in the TCM treatment group (ng/L: 363.67±97.83 vs. 225.87±47.71), and the differences were statistically significant (all P < 0.05). Conclusions Xiaozhong Sanjie decoction combined with Huayu powder external application can inhibit the levels of VEGF, IGF-Ⅰ and elevate the TGF-β1 level that possibly the mechanism of the combined therapy to shrink the thyroid nodule size.

Objective To observe the effects of Siweiyuganzi prescription on anti-peroxidation and blood lipid levels in experimental rats with hyperlipidemia. Methods Sixty male Sprague-Dawley (SD) rats were divided into normal control group, hyperlipidemia model group, Xuezhikang group, Siweiyuganzi prescription large, medium and small dose group according to the random number table method, with 10 rats in each group. The hyperlipidemia rat model was established by intragastric feeding with high fat emulsion everyday 10 mL·kg-1·d-1; normal saline 10 mL/kg was given to the normal control group, twice a day by intragastric feeding; 3 dosages of Siweiyuganzi suspended fluid 12.8, 6.4, 4.3 g·kg-1·d-1 intragastric administrations were given to Siweiyuganzi prescription large, medium and small dose groups respectively; Xuezhikang suspended fluid 0.3 g·kg-1·d-1 was given to Xuezhikang group intragastrically;the same volume of normal saline was given to hyperlipidemia model group. After 4 weeks, the level changes of blood lipid, serum superoxide dismutase (SOD), malonaldehyde (MDA), hydroxymethylglutaryl Coenzyme A (HMG-CoA) were observed. Results Compared to those in the normal control group, the levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), MDA, content and positive expression of HMG-CoA, alanine aminotransferase (ALT) were all higher in hyperlipidemia model group [TG (mmol/L): 6.59±0.72 vs. 4.32±0.36, TC (mmol/L): 7.10±0.25 vs. 5.98±0.40, LDL-C (mmol/L): 4.18±1.30 vs. 2.33±0.35, MDA (μmol/L): 26.05± 5.99 vs. 10.08±1.98, HMG-CoA content (ng/L): 54.60±2.90 vs. 48.73±3.09, HMG-CoA positive expression in liver tissue:(57.80±12.30)% vs. (22.00±4.92)%, ALT (U/L): 106.83±15.75 vs. 81.97±13.18]; SOD and high-density lipoprotein cholesterol (HDL-C) in hyperlipidemia model group were significantly decreased [SOD (kU/L): 295.47±37.51 vs. 345.13±19.76, HDL-C (mmol/L): 2.32±0.49 vs. 4.84±0.45, both P < 0.05]. Compared with the hyperlipidemia model group, the TG, TC, LDL-C, MDA, contents and positive expression of HMG-CoA in each group were significantly reduced, and the SOD and HDL-C were obviously increased, and the changes in the Siweiyuganzi high dose group were more significant than those of the Siweiyuganzi middle-and low-dose groups [TG (mmol/L): 4.70±0.46 vs. 5.40±0.31, 5.70±0.41, TC (mmol/L): 5.80±0.23 vs. 6.14±0.20, 6.56±0.32, LDL-C (mmol/L): 2.56±0.45 vs. 2.93±0.33, 3.28±0.32, HDL-C (mmol/L): 4.58±0.28 vs. 3.89±0.30, 3.59±0.08, SOD (kU/L): 381.45±20.68 vs. 360.60±30.16, 325.49±32.13, MDA (μmol/L): 16.98±5.39 vs. 17.89±5.37, 21.03±6.01, HMG-CoA content (ng/L): 50.58±0.77 vs. 52.16±0.66, 52.90±0.91, HMG-CoA positive expression in liver tissue: (27.90±6.03)% vs. (32.20±7.00)%, (43.00±8.39)%, all P < 0.05]. In the normal control group, there were positive cells scattered in the central vein area and loosely distributed around the portal area in the rat liver; in the hyperlipidemia model group, the positive cells were increased in the central vein area and the cells in relatively great number were seen around the portal area. While the positive cells in Xuezhikang group and in the high, medium and low dose Siweiyuganzi groups were decreased. Conclusion Siweiyuganzi prescription can regulate the levels of blood lipids, prevent and treat the lipid peroxidation caused by hyperlipidemia, and inhibit excessive expression of HMG-CoA in experimental rats with hyperlipidemia.