Objective To discuss human papilloma virus(HPV)prevalence and HPV genotypes distribution and the infection factors,to provide scientific guidance for the prevention and treatment of cervical cancer.Methods From March to November in 2009,605 women received cervical HPV testing in the Tumor Hospital of Harbin Medical University,to obtain specimens of cervical cytology,rapid flow-through hybridization technique (namely Hybribio flow-through hybridization)was used to detect HPV genotypes simultaneously.Single-factor and multivariate factors non-conditional Logistic regression analytic method was used to discuss the relationship between HPV infection of females and age,marital condition,level of education,level of income,occupation,initial age for sex,contraception,number of pregnancies,delivery approach and smoking.Results HPV infection rate was 21.49%(130/605),the positive rate of HPV infection in high-risk subtypes was 15.70%(95/605),the most common type was 5.29%(32/605)in the samples.Single factor non-conditional logistic regression model analysis showed that initial age for sex was the risk factor(X2=4.4618,P<0.05),HPV prevalence increased with a lower initial age for.sex reduced.But there was no significant difference in age,marital condition,education,income,occupation,contraception,number of pregnancy,delivery approach and smoking teams(X2=0.0525,1.8510,1.0348,0.2592,1.1176,1.5664,2.8835,1.4597,2.6161,all P>0.05).The analysis of multivariate factors nonconditional Logistic regression showed that the age of initially having sex,marital status and number of pregnancies were the risk factors(X2=21.6637,8.0574,15.7573,all P<0.05).Conclusions The risk factors for HPV infection are mainly about having sex too young,marital status and number of pregnancies,attention should be paid to screening for HPV.

OBJECTIVETo investigate the effects of hypoxia on lipid metabolism in the normal human hepatic cell line L02 and to investigate the underlying molecular mechanisms.

METHODSL02 cells were exposed to hypoxic conditions (experimental groups: at 1% O2, 5% CO2, 94% N2 for 3, 6, 12, 24, or 48 hours) or normoxic conditions (control group: at 21% O2). Lipid droplet accumulation and triglyceride content were measured in each group by oil red O staining and biochemical assay, respectively. The mRNA expression levels of hypoxia-inducible factor (HIF)-2a and sterol regulatory element binding protein (SREBP)-1c were detected by reverse transcription-polymerase chain reaction. Protein expression levels of HIF-2a, adipose differentiation-related protein (ADRP), and Fas were tested by Western blot analysis.

RESULTSLipid droplet accumulation and the triglyceride content were significantly higher in the hypoxia group than the normoxia group. In addition, the hypoxia groups had significantly down-regulated mRNA expression of SREBP-1c (12h: 0.236+/-0.043, 24 h: 0.287+/-0.044, 48 h: 0.342+/-0.049 vs. normoxia: 0.503+/-0.037; F = 28.37, P less than 0.01) and FAS protein (12 h: 0.562+/-0.054, 24 h: 0.674+/-0.062, 48 h: 0.682+/-0.057 vs normoxia: 0.857+/-0.069; F = 16.08, P less than 0.01). In normoxic cells, little or no expression of HIF-2a protein was detected by Western blot. In hypoxic cells, HIF-2a protein expression peaked at 6h (0.973+/-0.067). ADRP protein expression was significantly higher in hypoxia groups than in the normoxia group (12 h: 0.319+/-0.043, 24 h: 0.732+/-0.056 and 48 h: 0.873+/-0.066 vs. 0.211+/-0.019; all, P less than 0.05.

CONCLUSIONExposure to hypoxic conditions might induce lipidosis in normal human hepatic cells by stimulating HIF-2a and ADRP expression.

Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Hypoxia ; Cell Line ; Down-Regulation ; Hepatocytes ; metabolism ; Humans ; Lipid Metabolism ; Membrane Proteins ; metabolism ; Oxygen ; metabolism ; Perilipin-2 ; Sterol Regulatory Element Binding Protein 1 ; metabolism

OBJECTIVETo investigate the effect of Hepatitis B Virus X Protein (HBx) on the expression of lipid metabolism-related genes and its role in pathogenesis of hepatocyte fatty degeneration.

METHODSHepatitis B Virus X gene eukaryon expression vector pIRES2-eGFP-HBx was transfected into HepG2 cells to establish HepG2/HBx cell model for HBx expression. HepG2 cells transfected with pIRES2-eGFP (HepG2/pIRES2 cell) and non-transfected were used as controls. At 24, 48 and 72 hours after transfection, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the triglyceride(TG) content was detected. RT-PCR and Western blot were applied to detect the levels of sterol regulatory element binding protein-1 (SREBP-1), liver x receptor alpha (LXRalpha) mRNA and the levels of HBx, LXRalpha and fatty acid synthase (FAS) protein. At 24, 48 and 72 hours after transfection, the expression of GFP was found in HepG2/HBx and HepG2/pIRES2 cells, and increased gradually. The expression of HBx was detected only in HepG2/HBx cells, and was increased with time after transfection (F = 32.21, P less than 0.01). These suggested successful obtaining of HepG2-HBx cell model for HBx expression.

RESULTSAt 24h, 48h and 72h after transfection, the expression levels of LXRalpha mRNA (0.386+/-0.055, 0.505+/-0.071, 0.649+/-0.058 ) and SREBP-1 mRNA (0.395+/-0.055, 0.548+/-0.047, 0.795+/-0.058), as well as the levels of LXRalpha protein(0.178+/-0.036, 0.263+/-0.047, 0.347+/-0.058) and FAS protein(0.436+/-0.055, 0.608+/-0.053, 0.827+/-0.046) in HepG2-HBx group were dramatically higher than those in the controls at the same time points (all P less than 0.05/0.01), and were gradually increased with time (all P less than 0.05/0.01). A positive correlationship was observed between HBX protein level and the LXRalpha, SREbP-1 mRNA and LXRalpha, FAS protein levels. The difference of TG content between HepG2/HBx group and control groups was not statistically significant (P more than 0.05).

CONCLUSIONSHBx-LXRalpha-SREBP-1/FAS pathway suggested regulating transcription and expression of lipid metabolism-related genes, which might be one of the important molecular mechanism causing hepatocyte fatty degeneration.

Carcinoma, Hepatocellular ; metabolism ; Fatty Acid Synthase, Type I ; metabolism ; Hep G2 Cells ; Humans ; Lipid Metabolism ; genetics ; Liver Neoplasms ; metabolism ; Liver X Receptors ; Orphan Nuclear Receptors ; metabolism ; Sterol Regulatory Element Binding Protein 1 ; metabolism ; Trans-Activators ; genetics ; Transfection

Objective To investigate the expression and significance of CDK4 and cyclin D1 in gastric carcinoma,and to discuss their correlation with WHO histological classification,TNM stages and lymph node metastasis.Methods Forty paraffin wax specimens from patients with gastric carcinoma were collected and expression of CDK4 and cyclin D1 proteins were detected by immunohistochemistry.Results Immunohistochemistry demonstrated that the positive rates for CDK4 and cyclin D1 protein were 65% and 55%,respectively.There was positive correlation between CDK4 and cyclin D1 proteins(P0.05).Conclusion CDK4 and cyclin D1 proteins are frequently overexpressed in gastric carcinoma and there is a positive relationship between the two proteins,which may be involved in the development of gastric carcinoma.

The contribution of particles to cardiovascular mortality and morbidity has been enlightened by epidemiologic and experimental studies. However, adverse biological effects of the particles with different sizes on cardiovascular cells have not been well recognized. In this study, sub-cultured human umbilical vein endothelial cells (HUVECs) were exposed to increasing concentrations of pure quartz particles (DQ) of three sizes (DQPM1, <1 μm; DQPM3-5, 3-5 μm; DQPM5, 5 μm) and carbon black particles of two sizes (CB0.1, <0.1 μm; CB1, <1 μm) for 24 h. Cytotoxicity was estimated by measuring the activity of lactate dehydrogenase (LDH) and cell viability. Nitric oxide (NO) generation and cytokines (TNF-α and IL-1β) releases were analyzed by using NO assay and enzyme-linked immunoabsorbent assay (ELISA), respectively. It was found that both particles induced adverse biological effects on HUVECs in a dose-dependent manner. The size of particle directly influenced the biological activity. For quartz, the smaller particles induced stronger cytotoxicity and higher levels of cytokine responses than those particles of big size. For carbon black particles, CB0.1 was more capable of inducing adverse responses on HUVECs than CB1 only at lower particle concentrations, in contrast to those at higher concentrations. Meanwhile, our data also revealed that quartz particles performed stronger cell damage and produced higher levels of TNF-α than carbon black particles, even if particles size was similar. In conclusion, particle size as well as particle composition should be both considered in assessing vascular endothelial cells injury and inflammation responses induced by particles.

Objective: To observe the clinical efficacy of filiform fire-needling plus continuous passive motion (CPM) therapy for frozen shoulder. Methods: A total of 72 patients were randomized into an observation group and a control group by the random number table method, with 36 cases in each group. Patients in the control group received celecoxib capsule plus CPM, while those in the observation group received filiform fire-needling plus CPM. The whole course of treatment lasted for 2 weeks in both groups. The visual analog scale (VAS), Constant-Murley score (CMS) and range of motion of shoulder joint were measured for therapeutic efficacy evaluation. Results: The total effective rate was 91.7％ in the observation group, higher than 72.2％ in the control group, and the between-group comparison showed statistical significance (P<0.05). After treatment, the VAS scores in the two groups dropped significantly, the CMS as well as the range of motion including abduction, forward flexion and extension were all increased significantly, and the intra-group comparisons showed statistical significance (all P<0.05). The betweengroup comparisons showed the improvements in these items in the observation group were more significant than those in the control group (all P<0.05). Conclusion: Filiform fire-needling plus CPM can produce more significant efficacy than celecoxib capsule plus CPM for frozen shoulder; it can alleviate pain, improve shoulder function and restore joint range of motion in such patients.

The traditional culture methods and the molecular biology methods were used to study the rhizosphere bacterial diversity between fusarium wilt resistant and susceptible watermelon. The results showed that the diversity and the equality of cultured rhizosphere bacteria of resistant watermelon were higher than those of the susceptible watermelon. The reason was that the cultured rhizosphere bacterial di- versity index H′ and 1/D of the resistant watermelon were higher than those of the susceptible watermelon and that the cultured rhizosphere bacterial equality index E of the resistant watermelon were higher than those of the susceptible watermelon. The dominant cultured bacterial genotypes were different between re- sistant and susceptible watermelon. The genotype 1 is the dominant genotype of resistant watermelon, con- sists 51.1%. The genotype 7 is the dominant genotype of susceptible watermelon, consists 58.7%.

Objective To obtain elastic modulus through displacement of the ear structure. Methods The finite element model (FEM) of human ear structure based on Patran software was constructed and the neural network for inverse derivative of elastic modulus for the ear was established using Matlab software. The frequency response of the ear structure FEM was calculated to obtain the displacements of tympanic membrane and stapes. The displacements acting as input data of training samples and the corresponding elastic modulus acting as output data were used to train the neural network. Results The elastic modulus was inversely derived by adopting this mature neural network with relatively less error. Conclusions The viability of the proposed methods for inverse derivative of elastic modulus was demonstrated in this paper, which could provide a simple and effective method to obtain mechanical parameters for clinic work.

Purpose To study the clinical features, immunophenotypes and prognostic factors of primary breast diffuse large B-celllymphoma (PBDLBCL). Methods The clinical pathological data of 49 cases of PBDLBCL during January 2006 to December 2016 were retrospectively analysed, and the basic clinical and pathologic data, pathologic types and the immunohistochemical slides by EnVision method for staining were summarized. Results 47 cases were women and 2 cases were men. The age ranged from 24 to 79 year old with the median age of 48 year old. On microscopic observation, tumor cells were large to medium-sized which characterized as diffuse infiltration between the lobules of mammary gland, around the duct, interstitial and fat tissue, some were single file cord pattern. The immunophenotype showed 37 cases were of non-GCB, 12 cases were GCB type. Ki-67 index were greater than 40％. According to Ann Arbor staging, 16 cases were stage I EA, 28 cases were stage Ⅱ EA, 5 cases were stage Ⅳ E. IPI score: 30 cases with 0 ～1 score, 10 cases with 2 score, 9 cases with 3 score. Patients were followed up from 5 to 146 months, The 3-year overall survival (OS) rate was 51.2％ and 5-year OS rate was 36.7％, Single factor analysis showed that there were statistically significant difference in clinical stage, levels of LDH, IPI score, BCL-2 protein expression, and BCL-6 protein expression in 3 and 5 years of OS rate. The multiple factor analysis of Cox regression showed that the increase of IPI risk classification was the independent adverse prognostic factor of primary breast diffuse large B-cell lymphoma. Conclusion The diagnosis of PBDLBCL is confirmed by pathological biopsy and immunohistochemical markers. The immunophenotype was mainly non-GCB type. Comprehensive treatment including surgery, chemotherapy and radiotherapy is appropriate. The prognosis should be comprehensively evaluated by multiple factors. IPI increase risk classification is the independent adverse prognostic factor.

Objective To investigate the effect of acupuncture plus rehabilitation training on upper limb functional recovery in convalescent stroke patients.Methods One hundred and three convalescent stroke patients with upper limb motor dysfunction were randomized to a treatment group (54 cases) and a control groups (49 cases). Both group received conventional rehabilitation training. In addition, the treatment group received along-meridian syndrome differentiation-based acupuncture and the control group, sham acupuncture. Upper limb Fugl-Meyer Assessment (U-FMA) score, the Wolf Motor Function Test (WMFT) score, the modified Barthel Index (MBI) score and the 36-Item Short Form Health Survey (SF-36) score were recorded in the two groups of patients before and at the end of treatment and at the 3-month follow-up.Results In the two groups there were statistically significant differences in the U-FMA score, the WMFT score and the MBI score at the end of treatment and the 3-month follow-up compared with before (P>0.05 orP<0.01). At the end of treatment and the 3-month follow-up there were statistically significant differences in the U-FMA score, the WMFT score and the MBI score between the treatment and control groups (P<0.05 orP<0.01). The scores of SF-36 items except social functioning and bodily pain had statistically significant differences within the two groups at the end of treatment and the 3-month follow-up compared with before and between the two groups at the end of treatment and the 3-month follow-up (P<0.01 orP<0.05).Conclusion Acupuncture plus rehabilitation training can markedly improve upper limb function in stroke patients.