Objective To investigate the infection of HPV subtypes in male urethra in Xi'an area.Methods The eighteen subtypes of HPV DNA were detected in 478 cases of male patients by PCR-reverse dot hybridization assay (RDB) including low risk subtypes (HPV6,11,43) and high risk subtypes (HPV16,18,31,33) during 2015～2016.Results The total infection rate of HPV subtypes in 478 subjects was 44.97％ (215/478).The percents of subjects infected by one subtype,two ones,three ones,four ones and five ones were 32.85％ (157/478),8.79％ (42/478),2.09％ (10/478),0.42％ (2/478) and 0.84％ (4/478),respectively.The detection rates of low risk subtypes and high risk ones were 40.17％ and 22.38％ in which the most common subtypes were HPV6 (24.69％),HPV11 (12.13％),HPV16 (5.65％),HPV43 (3.35％) and HPV52 (2.30％) and those of others were from 0％ to 2.09％.The infection pattern of HPV subtypes gave priority to one subtype infection in all age groups.There was statistically different between H PVs (6,11,16,43,52,66) in twenty years or older subjects (x2 =12.879～109.7,P=0.000～0.025).Conclusion The majority of HPV subtypes detected in male urethra were HPV6,HPV11,HPV16,HPV43 and HPV52 in Xi'an area which provided epidemiological data for HPV infection in males.

Objective To investigate retrospective analysis on serological detection of hepatitis viruses in physical examination population.Methods Hepatitis A virus (HAV)IgM,HBsAg,HCV IgG,HDV IgM,HEV IgM and HEV IgG were detected by ELISA assay in physical examination population during 2009~2014.Results The positive rate of HAV IgM was 0.15%(2/1 315)in 1 315 physical examinees and that of HBsAg was 2.83% (3 360/109 965)in 109 965 ones during 2009~2014. The positive rates of HBsAg decreased trend in the last 6 years in general (χ2 =63.070,P =0.001).There was no signifi-cant difference for HBsAg positive rate among male physical examinees in last six years (χ2 =4.804,P =0.441),but the positive rate of HBsAg in female population decreased trend in the last 6 years in general (χ2 =18.046,P =0.003),moreo-ver the positive rate of HBsAg in male popolation was significantly higher than that in female ones (χ2 =126.9,P =0.000). The positive rate of HCV IgG was 0.79% (333/41 898)in 41 898 physical examinees during 2009~2014 and that decreased trend in last 6 years (χ2 = 18.380,P =0.003).There was no HDV IgM detected in 1 079 physical examinees serum.The positive rate of HEV IgM and IgG were 0.25% (3/1 195)and 3.93% (47/1 195)respectively in 1 195 physical examinees. Conculsion The physical examination population during 2009~2014 were mainly infected by HBV and HCV and the infec-tion rate decreased trendly in last 6 years,which provided important data for epidemiology of serological hepatitis viruses in Shaanxi provice.

Objective To establish serum NGAL reference range of healthy populations in Xi'an Area.Methods 2 665 cases (aged 6 to 95 years old,male 1 370,female 1 295) of health-check people were collected from March 2014 to October 2016 in Medical Examination Center of Shaanxi Provincial People's Hospital,and 682 cases (aged 0 to 6 years old,male 356,female 326) were collected from preschool children of prevention.Serum NGAL concentration of them were analysed by immunoturbidimetry method with the Automatic Biochemical Analysis Assembly Line of Beckman-AU5800,and the detection data for statistical analysis.Then established the reference range of serum NGAL population of different age and different sex in Xi'an.Results The serum NGAL levels in healthy subjects showed a skewed distribution,which were preschool children under 6 years of age 37.66±23.12 ng/ml,6～15 years 39.25±25.34 ng/ml,16～49 years 46.68±27.06 ng/ml,and 50～ 69 years 57.82±29.13 ng/ml.Compared the first two with the latter,there was a significant difference (t=0.589,P＜ 0.05).The serum NGAL levels of over 70 years were 61.87 ± 32.64 ng/ml,and there was a significant difference between the ages of 15 and 49 and over 70 years (t=8.529,P＜0.01).At the same time,the serum NGAL was closely correlated with age (r=0.298,P＜0.01).But there was no significant difference in serum NGAL level between male and female (t=0.263～0.542,all P＞0.05).87ng/ml was the upper limit of the reference value for the age of 50 years.Conclusion The level of serum NGAL was related to age and increased with age,but not with gender.

Objective To investigate the changes of the morphology,structure and biological characters of mutated Candida and through its genetic characteristics,research and reveal the mechanism of the variation at the molecular level.Methods Used different nutritional conditions,different growth conditions and different azole antifungal agents to induce mutation of the standard strains of Candida albicans.In clinical study,Candida mutant strains was isolated from vaginal secretions,pleu-ral effusion and gastric juice samples in patients of poor effect with Antifungal therapy,and studied on the morphological characteristics,and the nuclear structure,the biochemical reaction,the drug resistance,the bacterial composition and the ge-netic characteristics of above variants,etc.Results Mycelial?morphology:Candida were prone to mutation like bacteria, mutant bacteria could show G+ Aureus shape,G+ Bacillus,G+ long filamentous,G- Aureus shape,G- Bacillus and G- long filamentous;Nuclear structure:Candida mutant strains changed like prokaryotes under the electron microscope because it lost the original structure of eukaryotic cells.Biochemical reaction:there were 5 different items in 20 biochemical test ob-served.Drug sensitivity test:Candida mutated to antifungal drugs being originally sensitive was completely resistant,sensi-tive and resistant originally was completely sensitive,and the same as ordinary bacteria resistant.The cell component chan-ges:there was significantly different in Candida variant strain and the atavism of variant strain identified by mass spectrome-try.The most conservative fungalgene expression:Candida mutated had conservative gene expression of eukaryotes.It could be demonstrated that oidiomycetes mutant strains like bacterial morphology with prokaryotic cell biological characteristics was derived from Candida with eukaryotic cells.Conclusion Candida was prone to variation like bacterial morphology.The biological characteristics of mutant resembled prokaryote.There was a qualitative change among the standard strains of Can-dida albicans,mutant strains of oidiomycetes like bacterial morphology and the atavism of variant strain with clear genetic re-lationship under the electron microscope in the form of nuclear matter.The study on biological evolution,especially contact in prokaryotic cells and eukaryotic evolution has very important significance.

Objective:Analysis of anti-aging traditional Chinese medicine from the point of drug properties and explain the different types of drug characteristics.METHODS:The related literatures were derived from CNKI,and 51 traditional Chinese medicines were collected,then the ward method cluster analysis was carried out based on the property of a medicine data.Results:These drugs were divided into ten categories,the first kind of cold,bitter taste,liver meridian,can clear the liver;the second kinds of cold,bitter,spleen meridian,can clear splenopyretic;the third types of cold,sweet,nourishing yin;the fourth,five and six types were flat meridian,fiat sweet lung channel and flat sweet Spleen meridian,can nourishing liver,lung and spleen respectively;the seventh kinds of taste,to the kidney,can tonifying kidney-Qi;the eighth kinds of warm-natured,Spleen meridian,can fill the spleen meridian;the ninth kinds of warmnatured,spicy,to the kidney,tonifying kidney and dispelling cold;the tenth kinds of warm sweet to the kidney,tonifying kidney yang.Conclusion:The anti-aging traditional Chinese medicine was divided into ten categories based on the property of a medicine,the role of each type is different ways,reflecting the characteristics of anti-aging herbs.Adapt to the treatment of different aging patients.Through the classification of anti-aging drugs,it can be used to assist in the treatment of diseases,which provide clues for the study of anti-aging.

Three-dimensional (3D) printing is an additive manufacturing technology, which is widely used in automobile, aerospace, food, medicine and other fields. 3D printing technology brings novel solutions for precision medicine. In the field of hepatopancreatobiliary surgery, 3D printing is used in medical education, surgical simulation, patient-specific liver models printing in hepatectomy and liver transplantation. In the future, with the discovery and application of high-tech materials, 3D printing technology will be further developed in hepatopancreatobiliary surgery, and hepatobiliary surgery will usher in a new spring. This paper will review the application and future prospects of 3D printing technology in hepatopancreatobiliary surgery.

Objective:To investigate the role and mechanism of metformin in algesia of rats with type 2 diabetic neuropathic pain (DNP).Methods:Eighty sprague-dawley rats were randomly divided into normal control group ( n=15) and high-fat and high-glucose group ( n=65); normal diet and high-fat and high-sugar diet were given, respectively; before and 8 weeks after feeding, the body mass of rats and fasting blood glucose level were recorded, fasting insulin level was detected by ELISA, and insulin sensitivity index (ISI) was calculated. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) 8 weeks after feeding (baseline values) were measured in the high-fat and high-glucose group; after 12 h of fasting, intraperitoneal injection of streptozotocin (STZ, 35 mg/kg) was performed; 3 d after fasting, blood glucose was measured; 14 d after STZ injection, body mass was recorded and MWT and TWL were measured again: when MWT and TWL were ≤85% baseline values, it was defined that DNP model was successfully established ( n=45); and the left were into the diabetic painless group ( n=15). The rats with successful DNP were randomly divided into DNP group, DNP+vehicle group and DNP+metformin group ( n=15); 14 d after STZ injection, rats in the DNP+metformin group were given intraperitoneal injection of metformin (200 mg/kg) once daily for 14 consecutive d; DNP group did not accept any treatment, and rats in DNP+vehicle group were intraperitoneally injected with same amount of normal saline. MWT and TWL of all rats were measured 14 d after STZ injection, and 3, 7, 14 and 21 d after metformin injection. The expression levels of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α were detected by ELISA 7, 14 and 21 d after metformin injection. The fluorescence intensity of ionized calcium binding adaptor molecule-1 (Iba-1) in the spinal cord was detected by immunofluorescence staining, and the expression levels of Toll-like receptor 4 (TLR4), nuclear transcription factor (NF)-κB, phosphorylated (p)-NF-κB, adenylate activated protein kinase (AMPK), p-AMPK, and peroxisome proliferator activated receptor-γ coactivator (PGC)-1α in the spinal cord were detected by Western blotting 21 d after metformin injection. Results:(1) After 8 weeks of feeding, the body mass of rats in the high-fat and high-glucose group was significantly higher than that in the normal control group ( P<0.05); and the body mass of rats in the high-fat and high-glucose group was statistically lower than that in the normal control group 14 d after STZ injection ( P<0.05). Three d after STZ injection, the blood glucose level in high-fat and high-glucose group was significantly higher than that in normal control group ( P<0.05). After 8 weeks of feeding, the insulin level of high-fat and high-glucose group was statistically higher than that of normal control group, and the ISI in the high-fat and high-glucose group was significantly decreased as compared with that in the normal control group ( P<0.05). (2) As compared with those in the normal control group and diabetic painless group, MWT and TWL of DNP group and DNP+vehicle group were significantly decreased at each time point ( P<0.05). Three, 7, 14 and 21 d after metformin injection, MWT and TWL in DNP+metformin group were significantly increased as compared with those in DNP group and DNP+vehicle group ( P<0.05). (3) Seven, 14, and 21 d after metformin injection, the levels of IL-6, IL-1β and TNF-α in the spinal cord of rats in the DNP group and DNP+vehicle group were significantly increased as compared with those in the normal control group and diabetic painless group ( P<0.05); as compared with those in the DNP group and DNP+vehicle group, the levels of IL-6, IL-1β and TNF-α in the spinal cord of DNP+metformin group were significantly decreased ( P<0.05). (4) As compared with normal control group and diabetic painless group, the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+ vehicle group ( P<0.05). (5) As compared with those in the normal control group and diabetic painless group, the TLR4 and p-NF-κB protein expressions and p-NF-κB/NF-κB values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). As compared with those in the normal control group and diabetic painless group, the PGC-1α protein expression and p-AMPK/AMPK values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly decreased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly increased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). Conclusion:Metformin, by activating AMPK/PGC-1α signaling pathway, may inhibit the TLR4/NF-κB expression, reduce the activation of microglia and the expressions of pro-inflammatory factors, and thus alleviate DNP.

ObjectiveTo investigate the protective effect of folic acid against cholestatic liver injury in mice induced by bis（2-ethylhexyl） phthalate （DEHP） exposure and its mechanism. MethodsICR mice were randomly divided into control group， high-dose folic acid （H-FA） group， DEHP group， DEHP+low-dose folic acid （DEHP+L-FA） group， and DEHP+high-dose folic acid （DEHP+H-FA） group， with 6 mice in each group. The mice in the H-FA group， the DEHP+L-FA group， and the DEHP+H-FA group were given folic acid by gavage at the corresponding dose， and those in the control group and the DEHP group were given an equal volume of PBS solution by gavage. After 2 hours， the mice in the DEHP group， the DEHP+L-FA group， and the DEHP+H-FA group were given corn oil containing 200 mg/kg DEHP， and those in the control group and the H-FA group were given an equal volume of pure corn oil， by gavage for 4 weeks. Body weight and food intake were recorded every day， and blood and liver tissue samples were collected. A biochemical analyzer was used to measure the serum levels of total bile acid （TBA） and alkaline phosphatase（ALP）； HE staining was used to observe the histopathological changes of liver tissue； kits were used to measure the content of malondialdehyde （MDA） and superoxide dismutase （SOD） in the liver； LC-MS/MS was used to measure serum bile acid profiles； Western blot was used to measure the expression levels of proteins associated with hepatic bile acid metabolism. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group， the daily food intake of the mice in the DEHP group decreased significantly， and the body weight decreased significantly from day 10 （P<0.05）， and compared with the DEHP group， the DEHP+L-FA group and the DEHP+H-FA group had basically unchanged body weight and daily food intake （P>0.05）. Compared with the control group， the DEHP group had significant increases in liver weight index and the serum levels of TBA and ALP （all P<0.05）， with enlarged portal area， bile duct deformity and hyperplasia， and a small amount of inflammatory cell infiltration in liver tissue； compared with the DEHP group， the DEHP+L-FA group and the DEHP+H-FA group had a significant reduction in liver weight index （P<0.01）， and the DEHP+H-FA group had significant reductions in the serum levels of TBA and ALP （P<0.05）， with a significant improvement in liver histomorphology and structure after folic acid intervention. Compared with the control group， the DEHP group had a significant reduction in the content of SOD （P<0.05） and a significant increase in the content of MDA in the liver （P<0.01）， and compared with the DEHP group， the DEHP+H-FA group had significant reductions in the content of MDA and SOD （P<0.05）. Compared with the control group， the DEHP group had significant increases in the serum levels of α-muricholic acid （α-MCA），β- muricholic acid （β-MCA），deoxycholic acid （DCA）， lithocholic acid （LCA）， taurocholic acid （TCA）， taurodeoxycholic acid （TDCA）， tauroursodeoxycholic acid （TUDCA）， tauro-β-muricholic acid （T-β-MCA）， tauro-α-muricholic acid （T-α-MCA）， taurohyodeoxycholic acid （THDCA）， and taurolithocholic acid （TLCA） （P<0.05） and a significant reduction in ursodeoxycholic acid （UDCA）（P<0.05）； compared with the DEHP group， the DEHP+H-FA group had significant reductions in the serum levels of DCA， LCA， TCA， TDCA， TUDCA， T-β-MCA， T-α-MCA， THDCA， and TLCA （P<0.05）. Compared with the control group， the DEHP group had significant increases in the protein expression levels of FXR and CYP3A11 in the liver （P<0.01） and significant reductions in the protein expression levels of CYP7A1 and MRP2 （P<0.01）； compared with the DEHP group， the DEHP+L-FA group and the DEHP+H-FA group had significant reductions in the protein expression levels of FXR and CYP3A11 in the liver （P<0.05） and a significant increase in the protein expression level of MRP2 （P<0.05）， and the DEHP+H-FA group had a significant increase in the protein expression level of CYP7A1 （P<0.05）. ConclusionFolic acid has a protective effect against cholestatic liver injury in mice induced by DEHP exposure， possibly by regulating bile acid synthesis， catabolism， and transport and maintaining bile acid homeostasis.

OBJECT IVE To study the effects of ergosterol peroxide derivatives EP-3P on the proliferation ，migration and invasion of human tripe negative breast cancer cell MDA-MB- 231，and to provide reference for the development of breast cancer related drugs. METHODS MTT assay was adopted to detect the proliferation of MDA-MB- 231 cells after treated with 0（blank control），1.25，2.5，5，10，20，40 μmol/L EP-3P for 24，48 and 72 h. Wound healing assay and Transwell chamber method were adopted to detect the migration and invasion ability of MDA-MB- 231 cells after treated with 0（blank control ），5，10，20 EP-3P for 24 h. The apoptosis and cell cycle distribution were detected by flow cytometry. Western blot assay was used to detect the expressions of B-cell lympho ma-2（Bcl-2），Bcl-2 associated X protein （Bax），caspase-3，cleaved-caspase-3，cytochrome C （Cyt-C），matrix metalloproteinase- 2（MMP-2）and MMP- 9. RESULTS Compared with blank control group ，2.5，5，10，20，40 μmol/L EP-3P could significantly increase the inhibitory rate of cell proliferation （P＜0.05 or P＜0.01）in a dose and time- dependent manner. After 24 h treatment of EP- 3P（10，20 μmol/L），the rate of cell migration and the number of invasive cells were decreased significantly （P＜0.01），and cell was arrested at G 2/M stage （P＜0.05 or P＜0.01）；the apoptotic rate was increased significantly （P＜0.05）；the protein expressions of Bax ，Cyt-C and cleaved-caspase- 3 were upregulated significantly ， while those of Bcl- 2，caspase-3，MMP-2 and MMP- 9 were downregulated significantly （P＜0.01）. CONCLUSIONS EP-3P can inhibit the proliferation ，migration and invasion of human tripe negative breast cancer cells MDA-MB- 231 through mitochondrial mediated endogenous caspase pathway ，and induce the apoptosis of cells .

[Abstract] Objective: To observe the short-term efficacy and safety of Apatinib combined with radiotherapy and concurrent docetaxel and cisplatin chemotherapy in driver-gene-negative non-small cell lung cancer (NSCLC) patients with brain metastases. Methods: A total of 72 NSCLC patients with brain metastases, who were treated in our hospital from June 2018 to June 2019, were enrolled in this study. The driver gene was proved to be negative by next generation sequencing (NGS). The patients were divided into control group (36 cases) and treatment group (36 cases) by Digital random grouping method.The control group received 2 cycles of chemotherapy with docetaxel and cisplatin and concurrent radiotherapy for brain metastases, and the treatment group was given Apatinib anti-angiogenic treatment based on the regimen in control group. Primary study endpoints: confirmed objective response rate (cORR) and disease control rate (DCR); Secondary study endpoints: progression-free survival (PFS), quality of life (QOL) score, serum carcinoembryonic antigen (CEA), vascular endothelial growth factor (VEGF), and incidence of adverse drug events (AE). Results: Compared with the control group, cORR and DCR in treatment group were significantly improved [41.67% (15/36) vs 33.33% (12/36), 80.56% (29/36) vs 69.44% (25/36), all P<0.05], the median PFS was significantly prolonged (5.9 vs 4.6 months, P<0.05), and serum CEA and VEGF levels were significantly reduced [(16.5±2.3) vs (22.9±3.7) ng/ml, (291.6±42.6) vs (479.3±50.2) ng/L, all P<0.05], while the QOL score was slightly increased, but the difference was not statistically significant [(69.5±8.5) points vs (64.1±7.3) points, P>0.05]. There was no statistically significant difference in the incidence of acute brain edema, gastrointestinal reaction, bone marrow suppression, and liver dysfunction between the two groups of patients (all P>0.05); however, the incidences of oral mucositis, hand-foot syndrome, hypertension and proteinuria in the treatment group were significantly higher than those in the control group (all P<0.05). Conclusion: The efficacy of Apatinib combined with radiochemotherapy in driver-negative NSCLC patients with brain metastases is significantly better than that of radiochemotherapy alone, and the adverse reactions can be controlled. It is worthy of clinical recommendation.