Objective To investigate the effects ofviaminate on the proliferation and differentiation of HaCaT cells,a human keratinocyte cell line.Methods Cultured HaCaT cells were treated with various concentrations (2,5,10,15,20,25 and 30?g/mL) of viaminate for various durations.The cell proliferation was assessed by MTT method,the changes of cell cycle and apoptosis rate by flow cytometry,the changes of keratin 10 and involucrin mRNA expressions by semi-quantitative reverse transcription PCR.Results The proliferation of HaCaT cells was inhibited by the treatment with viaminate of≥2?g/mL for 48 h,and the inhibition rate was raised with the increase of treatment time and dosage.The viaminate of 30?g/mL inhibited the proliferation of HaCaT cells by 57.67% and 82.00% at 48 and 72 h after the incubation respectively,and elevated the mRNA expression of involucrin from 40.80% to 156.12%,decreased the mRNA expression of keratin 10 from 96.46% to 14.60%.The mRNA expression of involucrin increased with the elevation of viarninate dosage.Under the treatment with viaminate for 48 h,the cell population at G_1 phase significantly increased,that at S and G_2 phases decreased;the switching of G_1 to G_2 was inhibited;but the cell apoptosis was not affected.Conclusion Viaminate could inhibit the proliferation and induce the differentiation of keratinocytes.

Objective To analyze the distribution characteristics of blood uric acid and its relationship with the risk factors of metabolic syndrome in Uygur.Methods The questionnaire,anthropometric measurements,and biochemical detection were carried out in 4 428 healthy Uygur subjects in Xinjiang Urumqi and Kashi hospitals.Results (1) The prevalences of hyperuricemia and metabolic syndrome were 21.3 ％ and 8.2％,respectively.With the increased blood uric acid level,the incidences of coronary heart disease,hyperglycemia,hypertension,central obesity,and dyslipidemia were raised.Blood pressure,blood glucose,HbA1c,triglyceride,total cholesterol,apolipoprotein A,low density lipoprotein-cholesterol,body mass index (BMI),and waist to hip ratio (WHR) were increased with increased uric acid level,while high density lipoprotein-cholesterol was decreased.(2) The incidence of hyperuricemia was increased further when the number of metabolic syndrome components was accumulated (P＜0.01).With the increase of uric acid level,the prevalence of metablic syndrome gradually raised (P＜0.01).(3) Multiple logistic regression analysis showed that WHR (OR =7.639,95 ％ CI 1.744-33.466),coronary heart disease (OR =2.784,95 ％ CI 1.718-4.510),hyperuricemia (OR =2.155,95 ％ CI 1.457-3.188),smoking (OR =1.437,95％ CI 1.071-1.927),family history of metabolic diseases (OR =1.333,95％ CI 1.044-1.703),occupational pressure (OR =1.290,95 ％ CI1.021-1.631),and BMI (OR =1.146,95 ％ CI 1.112-1.181) were the risk factors of metabolic syndrome.Exercise (OR=0.472,95％ CI0.370-0.604) and low salt diet (OR=0.793,95％ CI0.662-0.949) were the protective factors.Conclusion Serum uric acid level is correlated with a variety of metabolic parameters.With the increased uric acid level,the risk of multiple metabolic abnormality was increased.Comprehensive prevention and control should be taken for the reduction of the risk factors and much attention should be paid to the adverse effects of hyperuricemia.

Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

OBJECTIVE: To study the disability identification for cases with clinical diagnosis of diffuse axonal injury (DAI) due to traffic accidents, and to explore the possible effects of DAI on identification results. METHODS: Five hundred and fifty-six cases of cerebral injury due to traffic accidents were collected, including 467 cases diagnosed with cerebral contusion or laceration and 89 cases diagnosed with DAI. The identification results of different groups with diagnosis of DAI diagnosis, diagnosis of DAI with cerebral contusion (laceration), and diagnosis of cerebral contusion or laceration without DAI were compared and statistically analyzed, based on the results of CT and MRI re-review. RESULTS: The disability identification levels in DAI group (20 cases), DAI group (69 cases) with cerebral contusion (laceration) and DAI group (467 cases) not complicated by cerebral contusion (laceration) were 7.72 +/- 1.09, 7.78 +/- 1.11, and 8.86 +/- 0.66, respectively. The disability levels of the two groups diagnosed with DAI were higher than those of the group without DAI diagnosis (P < 0.05). CONCLUSION Patients with DAI diagnosis might have more severe cerebral injury. In the identification process, one should pay attention to the possible missed diagnosis and misdiagnosis, and meanwhile avoid relying on those evidences provided only by CT and MRI.
Accidents, Traffic ; Brain Injuries/diagnosis* ; Diagnostic Errors ; Diffuse Axonal Injury/etiology* ; Disability Evaluation ; Forensic Pathology ; Humans ; Magnetic Resonance Imaging ; Resin Cements ; Tomography, X-Ray Computed

0.05).(4) Toxicity:Grade Ⅲ-Ⅳ myelosuppression was 60%(18/30)in Tp group,26%(8/31)in TC group and 30%(10/33)in PC group.The TP regimen had the greatest hematologic toxicity(P0.05). Conclusions As first line chemotherapy in epithelial ovarian cancer,TP regimen comparable to the standard chemotherapy regimen.

Objective To summarize the clinical manifestations of 46, XX ovotesticular disorder of sex development (DSD) caused by a NR5A1 heterozygous mutation. Methods The first case of 46,XX ovotesticular DSD was caused by a NR5A1 heterozygous mutation in China and was reported with a review of 11 similar cases in the literatures since July 2016. Results A 5. 6-year-old child raised as female was born with ambiguous genitalia. The left gonad was palpable in the inguinal region while the right one was located in abdomen. Gonadal histology showed both ovotestis. Vaginoscopy revealed a short, blind-ending vagina. No uterine was detected by laparoscopy. Repeated karyotype results were 46, XX with SRY gene negative. A heterozygous de novo mutation ( p. Arg92Trp) in the accessory DNA-binding region of NR5A1 gene was found in that child. Conclusions We reported for the first time in China a new phenotype caused by a NR5A1 heterozygous mutation-46,XX ovotesticular DSD. According to the review of literatures, such mutation seemed with incomplete penetrance. It could cause both 46, XX DSD and 46, XY DSD with varied manifestations. The possible underlying mechanism might relate to the impairment of the binding between the mutant protein and target DNA which might lead to a decreased inhibition of the male developmental pathway through downregulation of female antitestis genes.

BACKGROUNDAspergillus fumigatus (A. fumigatus) is a ubiquitous saprophytic fungus responsible for the majority of invasive mold infections in patients undergoing chemotherapy, organ transplantation or with persistent neutropenia. This study aimed to determine the role of E-cadherin for adhesion and endocytosis of A. fumigatus blastospores in the human epithelial cell line A549.

METHODSA. fumigatus blastospores were incubated with the total protein of A549 to investigate the binding of E-cadherin and blastospores followed by an affinity purification procedure. After establishing the adhesion model, the adhesion and endocytosis of A. fumigatus blastospores by A549 cells were evaluated by down-regulating E-cadherin of A549 cells using blocking antibody or small interfering RNA (siRNA).

RESULTSE-cadherin was adhered to the surface of A. fumigatus blastospore. Adhesion and endocytosis of the blastospores were reduced by blocking or down-regulating E-cadherin in A549 cells.

CONCLUSIONSE-cadherin is a receptor for adhesion and endocytosis of A. fumigatus blastospores in epithelial cells. This may open a new approach to treat this fungal infection.

Aspergillus fumigatus ; cytology ; Cadherins ; genetics ; metabolism ; Cell Line ; Endocytosis ; physiology ; Epithelial Cells ; metabolism ; microbiology ; Fungal Proteins ; chemistry ; metabolism ; Humans ; In Vitro Techniques ; Protein Binding ; physiology ; RNA, Small Interfering ; Spores, Fungal ; cytology

Background:Cerebellar fastigial nucleus (FN)is involved in regulation of visceral activities such as cardiovascular, ingestion,respiratory,and acute gastric mucosal injury,yet it is unclear whether it participates in the regulation of visceral hypersensitivity and what is the possible mechanism. Aims:To investigate the effect and possible mechanism of glutamic acid (Glu ) injection into cerebellar FN on chronic visceral hypersensitivity in rats. Methods: Chronic visceral hypersensitivity rat model was established by neonatal colorectal distension (CRD). After 8 weeks,the rats were divided into CRD group,solvent group (0. 2 μL 0. 9% NaCl solution injection into cerebellar FN),high-,medium-,low-dose Glu groups (12,6,3 μg Glu injection into cerebellar FN,respectively),3-MPA +Glu group (12 μg Glu injection after glutamate decarboxylase inhibitor 3-MPA injection into cerebellar FN),Bic + Glu group (12 μg Glu injection into cerebellar FN after GABAAreceptor blocker Bic injection into lateral hypothalamic area). Pain threshold,abdominal withdrawal reflex (AWR)score and abdominal external oblique muscle electromyography (EMG)were used to detect visceral sensitivity,and malondialdehyde (MDA)content and superoxide dismutase (SOD)activity were measured. Results:Chronic visceral hypersensitivity rat model was successfully established. Compared with CRD group,pain threshold was significantly increased (P＜0. 05),AWR score,EMG amplitude,MDA content were significantly decreased (P＜0. 05 ),and SOD activity was significantly increased in a dose-dependent manner in Glu group (P ＜0. 05 ). Compared with 12 μg Glu group,pain threshold was significantly decreased (P＜0. 05),AWR score,EMG amplitude, MDA content were significantly increased (P ＜0. 05),and SOD activity was significantly decreased in 3-MPA +Glu group,Bic+Glu group (P＜0. 05). Conclusions:Glu injection into cerebellar FN can significantly reduce the visceral sensitivity in rats. The mechanism may be that Glu in cerebellar FN produces GABA via glutamate decarboxylase,and then binding GABAAreceptor in lateral hypothalamic area,resulting in increased intestinal mucosal antioxidant capacity, thereby reducing visceral hypersensitivity.

Objective: To investigate the effects of glutamate (Glu) injected into hypothalamic paraventricular nucleus (PVN) on visceral pain of chronic visceral hypersensitivity (CVH) rats and its possible mechanism. Methods: Newborn SD rats were given CVH rat model by colorectal distension (CRD) on the 8th, 10th and 12th day after birth. Thirty rats with successful CVH model were randomly divided into CVH model group (CVH group), CVH + injection of saline into PVN group (NS group), CVH+ injection of Glu into PVN (3, 6, and 12 μg Glu, namely G3, G6, and G12, respectively), 6 rats in each group, and 6 SD rats with matching body mass were taken as sham operation group (Sham group). The pain behavior of the rats was evaluated by pain threshold, abdominal withdrawal reflex (AWR) score, and abdominal external oblique muscle electromyography (EMG). The expression of arginine vasopressin (AVP) and the proliferation of colon tissue were detected by immunohistochemical staining. The apoptosis of colon tissue was detected by TUNEL. Results: Compared with the NS group, the pain thresholds of the G3, G6 and G12 groups increased, and the AWR scores and EMG amplitudes decreased. The differences were statistically significant(Pain threshold: t=7.65, 16.31, 24.78, both P<0.05; AWR scores: t=-2.98, -4.77, -7.29, both P<0.05; EMG amplitudes: t=-3.06, -5.75, -8.92, both P<0.05). Compared with the Sham group, the expression of AVP in PVN of the CVH group and NS group decreased ((42.63±5.20) %, (18.67±2.94) %, (17.53±2.47) %; t=6.95, t=7.56, both P<0.05). The expression of AVP was increased after different doses of Glu into PVN, and the AVP level in G12 group ((18.15±6.49)%) was higher than that of NS group, the difference was statistically significant (t=-4.21, P<0.05). Compared with the Sham group, the expression of PCNA in colonic mucosal cells of the CVH group and NS group decreased ((65.48±1.68) %, (18.39±1.67) %, (17.69±1.68) %; t=34.35, t=34.80, both P<0.05). The expression of PCNA was increased after different doses of Glu injected into PVN, and the PCNA level in G12 group ((59.91±5.63)%) was higher than that of NS group, the difference was statistically significant (t=-12.44, P<0.05). Compared with the Sham group, the expression of apoptotic cells in colonic mucosal cells of the CVH group and NS group increased ((23.38±11.40)%, (83.79±3.57)%, (80.91±2.47)%; t=-8.77, t=-8.54, both P<0.05). The expression of apoptotic cells was decreased after different doses of Glu into PVN, and the G12 group was ((18.15±6.49) %). Compared with NS group, the difference was statistically significant (t=15.65, P<0.05). Conclusion Injection of Glu into hypothalamic PVN can alleviate the visceral pain behaviors in CVH rats, and its mechanism may be related to arginine vasopressin.