Ceramics ; Ergonomics ; Female ; Humans ; Male ; Occupational Exposure ; Workload

Objective To explore an effective method in reconstructing of burn scar contracture of the chin and neck in children. Methods The blood in the transverse cervical artery might flow reverse into the suprascapular artery and the posterior intercostals perforators through anastomic branches. An axial skin flap in the cervico-scapulo-dorsal area could be adequately supplied by this artery, and we had therefore used this flap to repair burn scar in the neck. Results 30 cases obtained satisfactory clinical results with this flap. The follow-up for two years in 18 patients showed that the neck of the patients moved about freely, and the develpment of the chin was not influenced, and the color of flaps was fine. Conclusion The blood supply of the flap is reliable and stable. So it is a very useful method in reconstructing the burn scar of the chin-neck in children.

Transcriptions are regulated by transcription factors. Natural transcription factors usually consist of at least two functional domains: a DNA-binding domain and an effector domain. According to this, novel artificial transcription factors are designed to up or down regulate transcription and expression of a target gene. The Cys2-His2 zinc finger domain is a DNA-binding module that has been widely used as the DNA-binding domain in artificial transcription factors. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. According to the "bipartite complementary" library strategy, a pair of zinc finger phage display libraries were constructed. After construction of the libraries, a 9bp sequence (5'-GCAGAGGCC-3') on the promoter of SV40 was chosen as a target for next step. After parallel selection, PCR amplification, desired fragments recovery, re-ligation, and additional rounds of selection, phage enzyme-linked ELISA experiments were performed to identify specific binding clones displaying the zinc fingers with predetermined sequence-specificity to our target sequence. Then two clones with strong ELISA signals were chosen to be tested for binding both to its full target site (5'-GCAGAGGCC-3') and to sites containing single transition mutations. The binding specificity of one of the two clones (clone 3) was shown to be fairly good. The three-finger DNA-binding domain targeted to SV40 promoter, that is, zinc finger sequences on clone 3, was fused to KOX1 suppression domain KRAB and cloned into pcDNA3.1 (+) (which expression product was artificial transcription factor). The zinc fingers (which expression product was the DNA-binding domain of artificial transcription factor) and KRAB domain only (which expression product was effector domain of artificial transcription factor) were also cloned separately into the same expression vector. All constructs contained an N-terminal nuclear localization signal. Every of the vectors (including pcDNA3.1 (+) without inserting sequences) were cotransfected with pGL3-Control and pRL-TK and the activity of luciferase was used to indicate the function of product from transfected expression vectors. Our artificial transcription factor was proved to repress the expression of reporter gene efficiently,while with only DNA-binding domain or effector domain the repression was not remarkable. By adding different effector domains and changing the DNA-binding domain, artificial transcription factor would have a wide range of potential applications.
Enzyme-Linked Immunosorbent Assay ; Genes, Synthetic ; genetics ; physiology ; Models, Theoretical ; Peptide Library ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Transcription Factors ; chemical synthesis ; chemistry ; metabolism ; Zinc Fingers ; genetics ; physiology

Human Zona Pellucida(ZP), which is a complex matrix surrounding oocytes,is comprised of three immunologically distinct glycoproteins(hZP1, hZP2 and hZP3). Because hZP3 possesses the sperm receptor activity and the acrosome-inducing activity, it has long been used as a candidate antigen to develop an immunocontraceptive vaccine. However, a large amount of native hZP3 protein is unavailable. It is an effective way to express hZP3 protein directly in vitro. Nevertheless, it had been reported that the rhZP3 protein produced in Pichia pastoris was not secreted but accumulated in the cells and could only be purified after being solubilized by strong denaturants. More unfortunately, after purification the final product required 6mol/L urea to maintain solubility. An improved project was advanced with the aim to express secreted and soluble rhZP3 protein in yeast. In this study, the fragment of hZP3 cDNA coding for aa 23 - 408, which the N-terminal leader was removed and most of the C-terminal transmembrane-like domain was reserved, was amplified by two PCR primers including EcoR I and Not I sites respectively and a His6 codon cassette was added to 5'-terminal. The hZP3 insert was incorporated into expression vector pPIC9K. The resulting recombinant yeast expression vector was designated pPIC9K-rhZP3. Linearized pPIC9K-rhZP3 was transformed into Pichia pastoris. After G418 selection, the recombinant Pichia pastoris strains were identified by PCR and the rhZP3 was expressed following the manufacturer' s protocol. Following induction with methanol, the rhZP3 protein was secreted and dissolved into the culture supernatant. SDS-PAGE and Western blot analyses showed that the apparent molecular weight of the expressed rhPZ3 proteins in yeast was smaller and a little size heterogeneity than native ones; after purified with Ni-chelating affinity chromatography, the final product's apparent molecular weight was about 32 - 34KD and their yield more than 20mg/L. We supposed that the C-terminal transmembrane-like domain be useful for secretion of rhZP3 into the culture supernatant and the expressed rhZP3 protein be incompletely digested by proteinases of Pichia into shorter fragments which all were glycosylated inhomogeneously. Fortunately, the fragments of rhZP3 protein can be recognized in Western blot by the polyclonal antibodies to porcine ZP3 which has showed a cross-reactivity with human ZP in vitro. It will be expected that the rhZP3 protein expressed in Pichia pastoris not only has immunogencity, say, it can rise antibodies in vivo to prevent spermatozoa-ovum binding, but also does not contain ovarian factors that might be the cause of undesired side effects, e.g. ovaritis and can be used as a safe immunogen in human antifertility vaccine research.
Blotting, Western ; Chromatography, Affinity ; Egg Proteins ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism ; Zona Pellucida Glycoproteins

Objective To investigate the prevalence of vitiligo in China through a multi-center and larse-population epidemiological survey.Methods A community-based survey was conducted in 6 cities from 6 provinces.Cluster sampling method was used.Subjects were required to fulfill the self-report questionnaires and received physical examination by dermatologists.EpiData and SPSS11.5 were utilized for statistical analysis. Results Totally,19 974 patients participated in this study,and 17 345 valid questionnaires were retrieved with a return rate of 86.84%.Of them,122 were found to have vitiligo.The prevalence and standardized prevalence of vitiligo was 0.70% and 0.56% in all patients,0.95% (75) and 0.69% in male patients and 0.50% (47) and O.45% in female patients.respectively.A significant elevation was observed in the prevalence of vitiligo in males than in females (P<0.01).The prevalence of vitiligo was increased with age and peaked in patients aging from 60 to 69 years and those over 70 years.The age at onset of vitiligo varied from 0 to 19 years in 21.85% of these patients,from 20 to 49 years in 47.05%.The most connnon type was focal vitiligo,which accounted for 36.06%,while the rarest type wag segmental type (2.46%).The pesitivity rate of family history of vitiligo was 9.84% in patients and 1.31% in community population (P<0.01).About 31.97% of the patients complained of negative influence of vitiligo on quality of life.Conclusions The standardized prevalence of vitiligo is 0.56%in 6 provinces from China.Males seem to have a higher prevalence of vifiligo than females.

OBJECTIVETo evaluate the efficacy of nucleos(t)ide analogues (NA) treatment and to assess the long-term outcomes, including survival, liver function improvement and virologic response, in patients with decompensated cirrhosis due to hepatitis B virus (HBV) infection.

METHODSPatients with Child-Turcotte-Pugh (CTP) scores more than or equal to 7, who had been treated with either lamivudine or other agents, but who were free of co-infection with other hepatitis virus were enrolled between January 2005 and December 2009. The study participants were subgrouped according to the antiviral drugs received or model for endstage liver disease (MELD) score for comparative analyses.Additionally, the 19 patients who were treated with NA for more than 5 years were investigated for changes in biochemical and virological indices, before and after the antiviral treatment.

RESULTSA total of 166 patients (125 males; 89 e-negative) and 52 untreated healthy patients (as control) were analyzed.The cohort of patients receiving antiviral therapy had significantly better 5-year actuarial survival than the untreated patients (74.1% vs.34.9%, P less than 0.001). For patients with MELD score more than or equal to 18, actuarial survival was not significantly different between the two groups (P=0.073).

CONCLUSIONAntiviral therapy significantly increases survival and improves the clinical long-term outcome of patients with HBV-induced decompensated cirrhosis.Antiviral treatment should be initiated at an early stage to maximize benefit in the improvement of clinical status.

Administration, Oral ; Antiviral Agents ; administration & dosage ; therapeutic use ; Cohort Studies ; Coinfection ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Lamivudine ; Liver Cirrhosis ; etiology ; Male ; Retrospective Studies ; Treatment Outcome

BACKGROUND: Increasing evidence has shown that visit-to-visit variability (VVV) of blood pressure (BP) is associated with an increased risk of cardiovascular disease (CVD). The objective of this study was to evaluate the impact of VVV of systolic blood pressure (SBP) and diastolic blood pressure (DBP) on the risk of CVD among patients with type 2 diabetes mellitus (T2DM) in China. METHODS: We conducted a retrospective cohort study of 10,163 T2DM patients who were not previously diagnosed with CVD from January 2008 to December 2012 in Ningbo, China. The VVV of BP was calculated using five metrics, including standard deviation (SD), coefficient of variation (CV), variation independent of mean, average real variability, and successive variability (SV) of measurements, obtained over a 24-month measurement period. Hazard ratios and 95% confidence intervals (CIs) were estimated by Cox proportional hazards regression models for the associations of variability in BP with risk of CVD. RESULTS: A total of 894 CVD events were observed during a median follow-up of 49.5 months. The hazard ratio in the highest quintile of SD of SBP was 1.24 (95% CI, 1.01 to 1.52) compared with patients in the lowest quintile. The association between higher VVV of DBP and risk of CVD was not consistent across different metrics and sensitivity analyses. CONCLUSION: Higher VVV of SBP was associated with an increased risk of CVD, irrespective of the mean SBP level. Future studies are needed to confirm these findings.
Blood Pressure ; Cardiovascular Diseases ; China ; Cohort Studies ; Diabetes Mellitus, Type 2 ; Follow-Up Studies ; Humans ; Retrospective Studies

OBJECTIVETo investigate the association between the development of hypertension and nutrition in school-age children in Fengdu County of Chongqing, China.

METHODSA total of 8 033 children from 2 public primary schools in Fengdu County of Chongqing, whose registered residence was in the subdistricts where the two schools were located, were selected as study subjects using cluster random sampling. Body height, body weight, and blood pressure were measured, and the semi-quantitative food frequency questionnaire was used for dietary survey. The association between body mass index (BMI), dietary nutrients, and the development of hypertension in children was analyzed.

RESULTSA total of 7 538 children were enrolled for analysis. The detection rates of obesity, overweight, and hypertension were 9.11%, 12.27%, and 11.83% respectively. In children with obesity and overweight, the detection rate of hypertension was 33.62% and 17.84% respectively, 4.02 and 2.13 times that in normal children. The multivariate logistic stepwise regression analysis revealed that increased intake of calcium and sodium increased the risk of hypertension (OR=1.003 and 1.002 respectively), while the increased iron intake and calcium intake per unit body weight reduced the risk of hypertension (OR=0.979 and 0.926 respectively).

CONCLUSIONSThe prevalence of hypertension and obesity in school-age children in Fengdu County of Chongqing is high. BMI and dietary nutrients are closely associated with the development of hypertension in children. Active control of body weight, adjustment of dietary structure, and limitation of sodium intake should be adopted to reduce the development of hypertension in school-age children.

Body Mass Index ; Child ; Child Nutritional Physiological Phenomena ; Female ; Humans ; Hypertension ; etiology ; Logistic Models ; Male ; Obesity ; complications ; Sodium, Dietary ; administration & dosage

The objective of this study was to investigate the effect of hexamethylene bisacetamide (HMBA) on differentiation of HL-60 cells and its possible molecular mechanism. HL-60 cells were co-cultured with different concentrations of HMBA (0.5, 1, 2 mmol/L) for 4 days, then the proliferation was assayed by MTT at different time points. Wright-Giemsa staining was used to observe the change in morphology. Cell differentiation antigen CD11b expression and the altered distribution of cell cycle in HL-60 induced by HMBA were analyzed by flow cytometry. The expressions of c-myc, mad1, p21, p27, hTERT and HDAC1 mRNA were detected by RT-PCR. The results indicated that the proliferation of HL-60 cells was inhibited by HMBA in a time-and-dose-dependent manner. Upon 2 mmol/L HMBA treatment, the HL-60 cells arrested at G(0)/G(1) phase and differentiated into granular line in morphology, with the up-regulation of CD11b expression. The expression of c-myc and hTERT mRNA obviously down-regulated, the expression of p21, p27 and mad1 mRNA up-regulated, while there was no change of the expression of hTERT mRNA. It is concluded that effect of HMBA on the differentiation of HL-60 cells may partly contribute to switch from c-myc to mad1 expression, to up-regulate expressions of p21 and p27 mRNA, and down-regulate hTERT mRNA expression, while there is no relation with the expression of HDAC1 mRNA.
Acetamides ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; genetics ; Cell Proliferation ; drug effects ; HL-60 Cells ; Humans

BACKGROUNDBerberine is one of the main constituents of Coptidis rhizoma (CR) and Cortex phellodendri. In this study, we investigated the beneficial effects of berberine on renal function and its possible mechanisms in rats with diabetic nephropathy (DN).

METHODSMale Wistar rats were divided into three groups: normal, diabetic model, and berberine treatment groups. Rats in the diabetic model and berberine treatment groups were induced to diabetes by intraperitonal injection with streptozotocin (STZ). Glomerular area, glomerular volume, fasting blood glucose (FBG), blood urea nitrogen (BUN), serum creatinine (Cr) and urine protein for 24 hours (UP24h) were measured using commercially available kits. Meanwhile, the activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) in serum, activity of aldose reductase (AR) and the expression of AR mRNA and protein in kidney were detected by different methods.

RESULTSThe results showed that oral administration of berberine (200 mg x kg(-1) x d(-1)) significantly ameliorated the ratio of kidney weight to body weight. Glomerular area, glomerular volume, FBG, BUN, Cr and UP24h were significantly decreased in the berberine treatment group compared with the diabetic model group (P < 0.05). Berberine treatment significantly increased serum SOD activity and decreased the content of MDA compared with diabetic model group (P < 0.05). AR activity as well as the expression of AR mRNA and protein in the kidney was markedly decreased in the berberine treatment group compared with diabetic model group (P < 0.05).

CONCLUSIONThese results suggested that berberine could ameliorate renal dysfunction in DN rats through controlling blood glucose, reduction of oxidative stress and inhibition of the activation of the polyol pathway.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Berberine ; pharmacology ; therapeutic use ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; drug therapy ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Wistar ; Streptozocin