Objective To investigate the distribution of Human Papillomavirus 16 (HPV16) infection and its mixed infection with other HPV subtypes in the Yunnan region. Methods 16 166 cases of women were tested using flow fluorescence Luminex technology. Results (1) HPV16 infection rate and mixed infection rate was 2.2%and 28.0%, respectively; (2) The most common type of HPV16 mixed infection was HPV52, followed by HPV33. The two kinds of mixed infection accounted for 39.8% of the total infection rate; (3) There was a significant difference between each age group of HPV16 mixed infection (Chi=26.39, <0.01) . Conclusion The HPV16 infection was mainly HPV infection in Yunnan region. HPV16 mixed infection merged mainly with HPV52 and HPV33. HPV16 mixed infection was associated with age.

Objective:To observe the influence of the “Warming yang and Nourishing qi” acupuncture treatment in the serum interleukin-12 (IL-12) and interleukin-18 (IL-18) expression levels,in the rats with experimental autoimmune myasthenis gravis (EAMG) and to analyze the mechanism of acupuncture treatment to myasthenis gravis (MG).Methods: AchRα1 129-145 peptide fragments were used to immune the female Lewis rats to establish EAMG rat models.30 successful EAMG model rats were randomly selected and divided into acupuncture treatment group ,drug control group and model control group ( n=10 );the unmodelled 10 rats purchased at the same period were used as blank control group.The rats in acupuncture treatment group were treated by “Warming yang and Nourishing qi” acupuncture treatment,30 min/time,once a day,7 d for a period,1 d rest between two periods,two periods of continuous treatment.The rats in drug control group were treated by the pyridostigmine lavaged treatment , 18.5 mg/( kg · d ) , continuous treatment of 15 d.The rats in the other two groups had no any special treatment.After treatment,the blood samples from the vein of tail root of the rats in 4 groups were collected ,and the serum expression levels of IL-12 and IL-18 were detected with ELISA method.Results:After treatment,compared with model control group ,acupuncture treatment and drug control group the serum IL-12 and IL-18 expression level of the EAMG rats were significantly decreased ( P<0.05 or P<0.01 ).Compared between acupuncture treatment group and drug control group showed that ,the serum IL-12 and IL-18 expression level of the EAMG rats were no significant difference ( P>0.05 ).Conclusion:“Warming yang and Nourishing qi” acupuncture treatment can reduce the serum IL-12 and IL-18 expression levels of EAMG rats to treat MG ,and the effect of treatment was similar to pyridostigmine.

Objective To explore the relationships between the expression of PECAM-1 and the degree of ALI in Paraquat induced lung injury model of rabbits. Method Thirty six adult New Zealand rabbits were randomly divided into three groups: 8 mg/kg (Group A), 16 mg/kg (Group B) and 32 mg/kg ( Group C) according to the dose of Paraquat which were infusion into stomach. After poisoned, the animals were monitored for seven days, and then sacrificed. The upper lobe of lung were removed for HE,Masson staining and immunohistochemisty. The ALl score, fibrosis of lung and expression of PECAM-1 were semiquantitative analyzed. Results Each group has 12 animals suffered from poisoning. The survival time of animals in Group C was (6. 47 ± 0. 99 ) days, shorter than (6. 09 + 1.04) days ( P = 0. 031 ) in Group B and (4. 77 + 2. 04) days ( P = 0. 0 07) in Group A. The ALI score were ( 8. 33 ± 1.03) points in Group A, superior to (9. 83 ± 1.17) points ( P = 0. 047 ) in Group B and ( 11.50 + 1.38) points ( P ＜ 0. 01 ) in group C, Group B vs Group C, P=-0.03o The fibrosis degree of lung was (31.09 +2.05)% in Group A,not severe as (34. 37 ±1.62)% (P=0. 002) in Group B and (36. 54 ±0. 44)% (P ＜0. 01 ) in Group C, Group B vs Group C, P = 0. 026. The Pearson correlation analysis showed the expression of PECAM-1 was negative correlated to ALI score (Coe = -0. 732, P =0. 001 ) and fibrosis degree of lung (Coe = -0. 779, P ＜ 0. 001 ) . Conclusions The expressions of PECAM-1 were significantly decreased in New Zealand after Paraquat poisoned, which were dose dependent, correlated to ALI scored and fibrosis degree of lung, so it may play an important role in the development of lung injured induced by Paraquat.

A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Objective To explore the effects of estrogen on oxidative stress of the lung tissue induced by acute paraquat (PQ) poisoning. Methods Thirty-two male adult New Zealand rabbits were randomly divided into model group and estrogen intervention group, 16 rabbits in each group. The model of lung injury induced by PQ poisoning was reproduced by feeding 16 mg/kg of 20% PQ through gastric tube. The rabbits in estrogen intervention group received intravenous infusion of 5 mg/kg estrogen after PQ challenge for 7 days, and the rabbits in model group received an equal volume of normal saline. Three rabbits in each group were sacrificed at 1, 2 and 3 days respectively after exposure. The lung tissue was harvested, the levels of reactive oxygen species (ROS) was determined by 2',7'-dichlorofluorescin diacetate (DCFH-DA), malondialdehyde (MDA) was determined by thiobarbituric acid (TBA), the mRNA expression of manganese-containing superoxide dismutase (MnSOD) was determined by reverse transcription-polymerase chain reaction (RT-PCR), and adenosine triphosphatase (ATP) content in mitochondrion was determined by enzyme linked immunosorbent assay (ELISA). The pathological changes in lung were observed under light microscopy using hematoxylin and eosin (HE) staining, and the lung injury was evaluated with lung injury score. Results The contents of ROS and MDA in lung within 3 days after PQ poisoning were gradually increased, and MnSOD mRNA expression and ATP content were gradually decreased. Estrogen intervention could significantly reduce the production of ROS and MDA after PQ poisoning [3-day ROS (fluorescence intensity): 161.05±30.04 vs. 188.30±31.80, 3-day MDA (mmol/L): 98.71±0.92 vs. 122.12±1.24], up-regulate MnSOD mRNA expression (integral A value: 3.05±0.90 vs. 1.22±0.24), and increase ATP content in mitochondrion (ng/L: 3.75±0.92 vs. 2.28±0.29) with statistically significant differences (all P < 0.01). In lung tissue after PQ poisoning, congestion, edema, focal pulmonary consolidation, pulmonary interstitial and alveolar space were infiltrated by a large number of neutrophil, alveolar interval were thickened obviously and the above phenomenon were most serious at 3 days after poisoning as shown under optical microscope. Estrogen intervention could significantly improve lung injury as compared with that of model group, and the lung injury score at 3 days was significantly lower than that of model group (11.8±0.7 vs. 13.5±1.0, P < 0.01). Conclusions The oxidative stress indicators in the lung tissue after PQ poisoning were obviously abnormal, the pathological damage was serious with time dependence. The administration of estrogen can reduce acute lung injury after PQ poisoning by reducing the oxidative stress.

Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time,and cell apoptosis was induced.Cell apoptotic rates were analysed by flow cytometry,and sensitivity of MML-1 cells to apoptosis was determined.The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3.Cyclin A,B_1 and E were selected as cell cycle markers for S,G_2/M and G_1 phase of MML-1 cells,and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h.After induction by Fas for 4 h,the active form of caspase-3 was mainly expressed in cells of G_1 phase,while rarely in cells of S and G_2/M phase.Cells with negative cyclin A and B_1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent.Cells entering into late G_1 and early S phase first express active form of caspase-3,and their sensitivity to Fas-mediated apoptosis is the highest.

A simple and accurate high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and evaporative light scattering detector (ELSD) was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation (ZFP).The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis (HCA),which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Objective To investigate the effects of the fibrin-derived peptide Bβ15-42 (FgBβ 15-42) on renal inflammation in acute kidney injury (AKI) induced by renal ischemia reperfusion (IR).Methods SD rats were randomly divided into sham group (the abdominal cavity were closed after separating the renal artery),IRI group (renal arteries of rats were occluded with microvascular clamps for 60 min),negative treated group (rats were injected with 3.6 mg/kg random peptide by tail vein) and FgBβ15-42 treated group (rats were injected with 3.6 mg/kg FgBβ15-42 by tail vein).Rats were sacrificed at 24 h or 48 h after reperfusion.Blood and kidney samples were collected and histological changes and renal function were examed.The mRNA and protein expressions of intercellular cell adhesion molecule-1 (ICAM-1) and interleukin-1β (IL-1β) were examined by immunohistochemistry,real-time PCR and Western blotting.Results Compared with sham group,Scr and BUN were obviously increased in IRI group (all P ＜ 0.05),pathologic changes of kidney were more serious (P ＜ 0.05).Compared with IRI group,in FgBβ15-42 treated group Scr and BUN were obviously decreased (all P ＜ 0.05),the injury of kidney tubulointerstitial was less serious (P ＜ 0.05).Compared with sham group,there was increased ICAM-1 and IL-1β in IRI group (all P ＜ 0.05),and they all peaked at 24 h.After treated with FgBβ15-42,the expression of ICAM-1,IL-1β were significantly decreased in kidneys compared to IRI group (all P ＜ 0.05).The above indexes had no significant differences between negative treated group and IRI group (all P ＞ 0.05).Conclusions FgBβ15-42 can protect kidneys against ischemia reperfusion injury in rats.The mechanism may be associated with down-regulated expressions of ICAM-1 and IL-1 β in the kidney.

Objective To explore the effects of pyrrolidine dithiocarbamate (PDTC) on the acute lung injury and the activation of Nrf2 pathway after Paraquat (PQ) induced lung injury.Methods Fortyeight adult male SD rats with lung injury induced by PQ were randomly (random number) divided into control group and PDTC group.Three animals were sacrificed at every 1-week interval,7d,14d and 21 days after PQ intoxication,and the lungs of rats were removed for acute lung injury score after HE staining,and for lung fibrosis assessment after Masson staining,and the levels of reduced glutathione (GSH) and malondialdehyde (MDA) in the lung tissue homogenate were assayed and the phosphorylation of Nrf2 (nuclear-E2-related factor 2) was detected by Weston blot.The mean values of detected variables between two groups were compared by t test,and survival curve was tested by Wilcoxon (Gehan) test.Results The intoxication symptoms of rats were obvious,and 4 rats in control group and 9 rats in PDTC group survived until 21days.The survival time of animals in PDTC group was longer than that in control group (Wilcoxon (Gehan) =10.17023,P =0.001).The levels of MDA in control group were higher than those in PDTC group,while the levels of GSH in control group were lower than those in PDTC group.The levels of phosphorylation of Nrf2 in PDTC group were higher than those in control group at 1-week intervals,1-week:(0.32±0.04) vs.(0.23±0.05),P=0.003; 2-week:(0.62±0.06) vs.(0.33±0.03),P＜0.001; 3-week:(0.61 ±0.04) vs.(0.33±0.05),P＜0.001.The acute lung injury (ALI) scores in PDTC group were lower than those in control group,1-week:(5 ± 0.95) vs.(8 ± 1.23),P =0.002 ; 2-week:(9±1.18) vs.(11±1.02),P=0.019; 3-week:(11±1.33) vs.(12±1.42),P=0.002.The percentages of lung fibrosis at 1-week intervals after PQ intoxication were (40.87 ± 7.25) ％,(43.38 ±5.71)％ and (45.91 ± 3.97)％ in control group,and they were higher than those in PDTC group (32.92±2.34)％,(33.45 ±3.04)％ and (35.27 ±3.81)％ in PDTC group,P=0.017,0.001 and 0.001 respectively.Conclusions Attenuation of acute lung injury and lung fibrosis,and prolongation of survival time of SD rats by PDTC were associated with activation of Nrf2 pathway.

Positive linear correlation was found between TSH level and detection rate of congenital hypothyroidism (CH). When measured with a cut-off point at 9, 10, 15 and 20 mU/L of TSH, the sensitivity of positive detection rate for CH was 99.77%, 96.80%, 81.25% and 71.88%, respectively. TSH > 9 mU/L seems to be a reasonable cut-off value.