Objective: To construct a lentiviral vector (LV) of RNA interference (RNAi) targeting HBs gene and to observe its effect on the replication of HBV and expression of antigens. Methods: The effective sequence of siRNA targeting HBs gene was confirmed in our previous study. The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed, synthesized and cloned into the lentivirus vector (pGCLM-GFP). The resulting lentivirus vector containing HBs shRNA was named LVshHBs, and it was confirmed by PCR and sequencing. 293T cells were cotransfected with lentivirus vector pGCLM-GFP, pHelper 1.0 and pHelper 2.0. All virus stocks were produced by calcium phosphate-mediated transfection. The titer of virus was tested according to the expression level of GFP. HepG2. 2.15 cells were infected with LVshHBs and the supernatant of the cells was subjected to ELISA, Western blotting analysis and HBV DNA quantitative analysis. Results: PCR and DNA sequencing demonstrated that the LVshHBs-producing HBs shRNA was successfully constructed. The titer of concentrated virus was 5 × 108-2 × 109 TU/ml. The inhibitory effect was efficient and the corresponding viral transcript and expression of antigens were decreased after infection. The inhibitory effect was observed 4 days after infection and peaked 9 days after the initial treatment with RNAi. Secreted HBsAg was reduced by > 70% in cell culture compared with the negative control, which is also confirmed by Western blotting and real time PCR. After quantification of HBV DNA, the level of DNA relative to the controls was also significantly reduced after RNAi treatment(P<0.05). Conclusion: The lentivirus RNAi vector of HBs has been successfully constructed. The lentiviral microRNA-based RNAi targeting HBs can specifically mediate the down regulation of HBs expression, inhibiting HBV replication and antigen expression.

Objective To observe the damage to Langerhans cells induced by UVB irradiation,and to evaluate photoprotective effect of these cells from UVB irradiation by epigallocatechin-3-gallate (EGCG).Methods Biopsy specimens were obtained from normal adult foreskin,and epidermal cells were isolated.Density gradient centrifugation and magnetic cell sorting were used simultaneously to purify Langerhans cells from the cell suspension.These cells were then divided into three groups,control (no ir- radiation or EGCG treatment),UVB (irradiation) and EGCG (irradiation+ECCG treatment) groups. The cells in the UVB and EGCG group were irradiated by UVB (30 mJ/cm~2).After the irradiation,the U- VB group was incubated with RPMI-1640 containing 10% bovine serum for 4 hours,while the EGCG group with the same medium containing 200?g/mL of EGCG for 4 hours.Another four hours after the treatment, the cells were collected for the detection of apoptosis rate by propidium iodide staining and flow cytometry. Results Exposure to UVB (30 mJ/cm~2) significantly increased the apoptotic rate of Langerhans cells.The apoptotic rate in EGCG group was significantly lower than that in the UVB group,but was higher than that in the control group.Conclusion Rate of apoptosis of Langerhans cells could be increased by UVB irradia- tion,while EGCG could prevent the increase of apoptosis.

Renal transplant (RT) recipients have a high risk of developing cardiovascular diseases.However,the effects of renal transplantation on the development of arteriosclerosis have been controversial.The carotid intima-media thickness (CIMT) and diameter (CD) are important indicators of vascular remodeling and arteriosclerosis.In this study,31 patients with hemodialysis (HD),31 RT recipients and 84 age-and gender-matched control subjects were enrolled.Their CIMT and CD were measured by ultrasonic radiofrequency tracking,and the linear regression models and Z test were used to identify the progression of arteriosclerosis and the risk factors.Compared with HD group,RT group had significantly lower CIMT and CD.CIMT was found to be associated with age,body weight,resistance index and diastolic velocity,while CD was associated significantly with age,body weight,pulsatility index,end diastolic velocity and diastolic blood pressure (DBP),respectively.The correlation curves between CIMT and age showed the slopes of curves were decreased successively in control,RT and HD groups,and the curves between CD and age showed the slopes were decreased in order of RT ＞ control ＞ HD groups.It was concluded that CIMT and CD were significantly correlated with age in RT and moderately with age in HD patients.RT could reduce the progress of arteriosclerosis in patients with end-stage renal disease.

Objective To explore the relationship between the endemic arsenism and the liver,renal damage.Methods Some permanent residents were selected as investigated subjects who lived at 3 villages in Datong in Shanxi Province,an arseniasis-endemic areas,These objects were divided into arsenic poisoning and control group on the basis of Diagnosis Standard for Endemic Arsenism(WS/T 211-2001).Then blood and urine samples were collected in the surveyed people.Serum glutamate pyruvic transaminase(ALT)were detected by Enzyme-linked immunosorbent assay as the indicator of the impaired hepatic function.The microdosis albumen (mAlb)and acetylglucosaminidase(NAG)in urine were detected by end-point method and alkaline picric acid as the renal damage indicators.Results A total of 661 people investigated,of which 144 cases were arsenic poisoning patients.The rates of abnormal liver function were significant hisher in arsenic poisoning group[10.42% (15/144)]than that in control[5.22%(27/517)],and both wag significant[X2=5.107,P＜0.05;OR=2.11,95%CI (1.09-4.08)].The geometric mean of mAlb/Ucr was 2.16 mg/g Cr in control,and 2.31 mg/g Cr in arsenic poisoning group,and both was not significant(t=-1.71,P＞0.05).The geometric mean of NAG waft higher in arsenic poisoning group(2.43 U/g Cr)than that in the control(2.22 U/g Cr),and both was significant(t=-3.55, P＜0.05).Conclusions The damage of the liver and renal function were related with endemic arsenism,and NAG is the early indicators suggesting impaired renal function due to endemic arsenism.

Objective To investigate the prevalence for endemic fluorosis of drinking water type and to discuss the relationship between endemic fluorosis and urinary fluorine in Linyi county, Shanxi province. Methods In 2006, three counties were selected as heavy, medium and control areas according to the distributing feature of the disease. The dental fluorosis in each spots was examined by Dean method. The levels of urinary fluorine were determined by fluorine selective ion electrode. The skeletal fluorosis of adults were examined by X-ray. Results There was evident differences of dental fluorosis and skeletal fluoresis among the heavy and the medium endemic fluorosis and control areas(X~2 = 410.945, P < 0.01 ), the prevalence of dental fluoresis in the medium area and the heavy area were 92.34% (253/274), 90.09% ( 291/323), significantly higher than in the control area[23.27% (64/275), X~2 = 274.927,268.287, all P < 0.01]. The heavy area had the highest rate of the skeletal fluorosis rate [59.75% (141/236) ], the medium area had the middle-level of the skeletal fluorosis rate[24.76%(52/210), X~2 = 183.578, P< 0.01]. Urine fluorine contents in both beavy[ (4.69 ± 0.17)mg/L] and medium areal (4.86 ± 0.13)mg/L] were higher than that in the control areas[ (1.75 ± 0.04)mg/L, H = 411.197, P< 0.01], and there was linear relevance between the different degree of skeletal fluorosis and urine fluorine contents (r = 0.508, P < 0.01). Conclusions The local fluoresis condition of Linyi county in Shanxi province was serious. The degree of skeletal fluorosis is associated with the fluoride content in urine.

BACKGROUNDPrevious studies suggested that zinc level was related to a certain diabetic microvascular complication. However, the relationship between zinc level and all the microvascular complications in type 2 diabetic patients remains unknown. The purpose of this study was to analyze the relationship between zinc level and each diabetic microvascular complication and identify the features related to low serum zinc level.

METHODSWe included the hospitalized patients with type 2 diabetes (T2D) at our department from May 30, 2013 to March 31, 2014. We initially compared the serum zinc levels between patients with specific microvascular complications and those without. We then analyzed the association between zinc level and each microvascular complication. Furthermore, we identified the unique features of patients with high and low serum zinc levels and analyzed the risk factors related to low zinc level.

RESULTSThe 412 patients included 271 with microvascular complications and 141 without any microvascular complications. Serum zinc level was significantly lower in patients with diabetic retinopathy (P < 0.001), diabetic nephropathy (DN, P < 0.001), or diabetic peripheral neuropathy (P = 0.002) compared with patients without that specific complication. Lower zinc level was an independent risk factor for DN (odds ratio = 0.869, 95% confidence interval = 0.765-0.987, P < 0.05). The subjects with lower serum zinc level had manifested a longer duration of diabetes, higher level of hemoglobin A1c, higher prevalence of hypertension and microvascular complications, and lower fasting and 2-h C-peptide levels.

CONCLUSIONSLower serum zinc level in T2D patients was related to higher prevalence of diabetic microvascular complications, and represented as an independent risk factor for DN. Patients with lower zinc level were more likely to have a longer duration of diabetes, poorer glucose control, and worse β-cell function.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; blood ; complications ; Diabetic Nephropathies ; blood ; etiology ; Diabetic Neuropathies ; blood ; etiology ; Diabetic Retinopathy ; blood ; etiology ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Zinc ; blood

Objective: To explore the effect of the umbilical cord mesenchymal stem cells(UC-MSCs) on the pulmonary fibrosis in silicosis rats. Methods: SPF male Sprague Dawley rats were randomly divided into control group, silica model group and UC-MSCs treatment group with 12 rats each group. SiO₂ intra-tracheal injection(0.5 ml of 50 mg/ml/rat) were applied to silica model group and UC-MSCs treatment groups. After that UC-MSCs treatment group received 1 ml UC-MSCs suspension (3×10⁶ cells/ml) by tail vein injection on the 29th, 36th, 43th and 50th day after exposure to the first silica suspension. On the 60th and 75th day after exposure to silica suspension, all animals were examed for pulmonary CT. Then the rats were euthanized on 75th day after the first exposure to silica.Lung's histopathological examination of the rats from all the groups were carried out. The content of hydroxyproline in lungs, TGF-β1 and IL-6 in serum were examined. Results: The lung's histopathological examination showed no obvious inflammatory cell and no fibrosis in the lung tissue of the control group, there were a lot of inflammatory cell aggregation and collagen fiber deposition in silica model group, while in the UC-MSCs intervention group and treatment group, there were less inflammatory cells and collagen fiber. The rats from silica model groups had higher HYP, TGF-β1 and IL-6 than the rats from UC-MSCs treatment group and control group. Lung fields of rats in the control group were clear and no obvious high-density shadow. Different-sized granular high-density shadows or reticular fibrous shadows were found diffusely distributed in the lungs of the rats in silica model group. Lung field of rats in UC-MSCs intervention group and treatment group were less high density shadows, and more clear. Conclusion UC-MSCs can alleviate the pulmonary fibrosis in silica model rats through regulating the secretion of some fibrosis related cytokines.

Objective:To describe the epidemiological characteristics of mental disorders in Ximeng Wa Au-tonomous County and explore psychosocial risk factors of mental disorders. Methods:Two thousand three hundred and eighty one residents aged 1 8 years and over were sampled using multistage sampling in Ximeng Wa Autono-mous County in 2013. All respondents were investigated by face-to-face interview. Mental disorders were screened out by using the General Health Questionnaire (GHQ)and diagnosed according to the International Classification of Diseases Checklist (ICD-10-Checklist)criteria. Results:Life time prevalence of any mental disorder was 19. 86%(419/2110). The lifetime prevalence rates of substance use disorder,anxiety disorder,insomnia,mood disorder, schizophrenia were 12. 99%(274/2110),6. 30%(133/2110),2. 94%(62/2110),2. 32%(49/2110)and 1. 00%(21/2110)respectively. Male (OR=0. 43),older age (35 -49 years,OR=1. 78;50 -64 years,OR=2. 59;≥65 years,OR=3. 5 1 ),unmarried and other marital status (OR=0. 3 1 ),non-Wa and non-Lahu ethnic groups (OR=0. 29)were associated with neurotic,stress-related and somatoform disorders. Male (OR=2. 41),older age (35 -49 years,OR=2. 29;50-64 years,OR=3. 20;≥65 years,OR=4. 58),non-farmer and non-self-employed occupation (OR=0. 41),and non-Wa and non-Lahu ethnic groups (OR=0. 32)were associated with psychoactive substance use disorder. Male (OR =0. 35 ) and order age (≥65 years,OR =3. 05 ) were associated with mood disorders. Conclusion:Lifetime prevalence of any mental disorder,substance use disorders and anxiety disorders are high in Ximeng Wa Autonomous County. Measures should be strengthened against prevalence of mental disorders in ethnic minority areas.

Adult ; Asian Continental Ancestry Group ; DNA Mutational Analysis ; Female ; Humans ; Male ; Middle Aged ; Parkinsonian Disorders ; genetics ; Pedigree ; Proton-Translocating ATPases ; genetics ; Young Adult

BACKGROUNDThe Akt2 protein kinase is thought to be a key mediator of the insulin signal transduction process. Akt2 is suggested to play a role in glucose metabolism and the development or maintenance of proper adipose tissue and islet mass. In order to determine whether the Akt2 gene plays a role in the pathogenesis of type 2 diabetes characterized by insulin resistance, and to further identify if variations in this gene have a relationship with type 2 diabetes, we sequenced the entire coding region and splice junctions of Akt2 and made a further case-control study to explore the association between single-nucleotide polymorphisms (SNPs) in this gene and type 2 diabetes in the Chinese Han population.

METHODSWe selected 23 probands with a type 2 diabetic pedigree whose family members' average onset age was within 25 to 45 years old. The body mass index of all the participants was lower than 28 kg/m(2) and all of them were insulin-resistant (the fasting insulin level > 100 pmol/L or 16 µIU/ml). The entire coding region and splice junctions of Akt2 were directly sequenced in these 23 probands. SNPs with a frequency of minor allele over 20 percent were selected to be further studied in a case-control study. We chose 743 non-diabetic subjects as the control group and 742 type 2 diabetic patients as the case group. All these subjects were genotyped. A Snapshot Technology Platform (Applied Biosystems) was used for genotyping.

RESULTSThe Akt2 genes from all 23 subjects were successfully sequenced. We did not identify any mutation in the type 2 diabetic pedigree. Two SNPs were identified, 13010323T > C and 13007939G > T. 13010323T > C was in intron 9, which was the location of rs2304188 reported in Genbank. Its minor allele frequency was 13.04%. 13007939G > T was in the 3'-untranslated region (UTR) of exon 14, which was the location of rs2304186 reported in Genbank. Its minor allele frequency was 34.78%. The allele frequency of rs2304188 and rs2304186 were consistent with the frequency reported in Genbank. In the case-control study with 742 patients and 743 controls, there was no significant difference between the two groups for the allele frequency of rs2304186 (odd ratio: 0.96, 95% confidence interval: 0.82 - 1.12, P = 0.597).

CONCLUSIONSThe Akt2 gene is not a major cause of diabetes in a non-obese Chinese Han population characterized by insulin resistance. There is no significant relationship between rs2304186 and type 2 diabetes in the Chinese Han population.

Adult ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Proto-Oncogene Proteins c-akt ; genetics