Objective To investigate the protective effect and mechanism of MST1 inhibition on kidney tissue in diabetic rats,and to find a new therapeutic target for diabetic nephropathy.Methods Total of 54 male SD rats enrolled in this study were divided into 3 groups including normal control (group A,n=18),MST1 inhibition group (Group B,n=18) and diabetes group (group C,n=18).Diabetes was induced by a single streptozotocin (STZ,50 mg/kg) injection in group B and group C.rats in group B received lentiviral vector contain Mst1 interference RNA (shRNA) and the rats in group C received empty vector.The end of 4th,8th and 12th week after modeling were considered as time points in this study.At each time point,the level of 24 hours urine protein (24-HUP),blood glucose and serum creatinine were examined.Pathological changes were observed with HE stain; Injury of podocyte and glomerular basement membrane (GBM) were examined with transmission electron microscope (TEM).The intensity and location of MST1 in kidney tissue were detected by immunohistochemistry.The level of MST1,Phosphorylated-MST1,nephrin,Caspase-3 and FasL were detected by western bloting.Results (1) At the starting point,there were no significant differences among groups in terms of weight,activity,eating and drinking.Since the end of 72nd hour after modeling,the levels of glucose in both group B and group C,compared to those in group A,significantly increased (P ＜ 0.05).There was no significant difference between group B and group C for glucose level at each time point (P ＞ 0.05); the level of 24-HUP increased significantly since the end of 4th week after modeling,and the level in group C was higher than its counterpart in group B at the same point (P ＜ 0.05); (2) There was no significant pathological lesion observed in group A.Without obvious K-W nodular changes,mesangial proliferation was observed in group B and group C.It was shown by TEM that podocyte fusion and thickening of the GBM could be found in group B and group C.The pathological change in group B was better than that in group C; (3) Compared to group A,it was shown by western blot that the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B and group C were significantly higher (P ＜ 0.05),and the levels of nephrin in group B and group C were significantly lower (P ＜ 0.05) since the end of 4th week after modeling.Meanwhile,the levels of MST1,Phosphorylated-MST1,Caspase-3 and FasL in group B were significantly lower than that in group C at each time point (P ＜ 0.05),the level of nephrin in group B was significantly higher than the one in group C; (4) It was shown by immunohistochemistry that there was low MST1 expression in normal condition,especially in cytoplasm of tubular epithelial cells.The level of MST1 in group B and group C significantly increased after modeling,and the change could be the same as Western blot shown.Conclusions MST1 pathway could be involved in kidney injury induced by diabetes.MST1 inhibition could alleviate the kidney injury in STZ-induced diabetes animal model.

OBJECTIVE: To study the regulatory effect of IL-35 on the balance of Treg/Th17 cells in AR patients. METHOD: In this study, 30 cases were randomly selected from outpatients of otolaryngological department in the second hospital of Hebei Medical university who were diagnosed as AR. Another 20 healthy cases enrolled from physical examination branch of our hospital were control group. The expression level of IL-35 and IL-17 in peripheral blood were detected by using ELISA and defeced CD4+CD25+Foxp3+ T cell and CD4+IL-17+T cell expression level were identified via flow cytometry. RESULT: The expression level of IL-35 in AR group was obviously lower than that in control group, and the difference was a statistically significance (t = -8.145, P < 0.01). The expression level of IL-17 in AR group was obviously higher than that in control group, and the difference was a statistically significance (t = 14.969, P < 0.01). There was a remarkable negative correlation between the IL-35 and IL-17 expression in the serum of AR group (r = -0.773, P < 0.01). The percentage of CD4+CD25+Foxp3+ T cell in CD4+ T cell was significant lower in AR group than that in control group (t = -13.678, P < 0.01). The percentage of CD4+IL-17+ T cell in CD4+ T cell was much higher in AR group than that in control group (t = 5.632, P < 0.01). There was a remarkable negative correlation between the Treg and Th17 expression in the peripheral blood of AR group (r = -0.613, P < 0.01). There was a positive correlation between the expression of CD4+ CD25+Foxp3+ T cell and IL-35. There was a negative correlation between the IL-35 and Th17 in AR group (r = 0. -594, P < 0.01). CONCLUSION The lower expression of IL-35 was related to the incidence of AR, and it was an important cytokines for that. The lower expression of IL-35 may inhibit the proliferation of Treg cells, lead to hyper function of Th17 cells, increase secretion of s IL-17 and result in unbalance of Treg/Th17 cells; these may be the important mechanism of the occurrence of AR, thus regulation of IL-35 may become a new target for the immunological therapy of AR.
Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-17 ; blood ; Interleukins ; Rhinitis, Allergic ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

Objective To explore the best time point for the ipsilateral hepatocellular apoptosis and the contralateraI hepatic hypertrophy after selective portal vein embolization(SPVE)in rabbit.Methods In a randomized study design,forty rabbits were divided into 5 groups with 8 rabbits per-group,including one as the control and the other 4 were treated with SPVE during open surgery.The rabbits were killed postoperatively,in 3,7,14,21 days respectively after the embolization.The hepatic lobes volume,the ipsilateral hepatocellular necrosis rates and apoptosis index,and liver functions were determined as well. Results In the treatment groups,the average amount of the right liver volumes decreased from 46.4 cm~3 preoperatively to 46.0,44.4,42.0,39.7 cm~3 in groups of 3,7,14,21 days postoperatively;meanwhile,the left liver volumes increased from 54.0 cm~3 preoperatively to 54.5,56.3,61.7,63.9 cm~3 respectively during 3, 7,14,21 days after the procedures.The rates of future remaining live volumes(FRLV)increased from 53.8% preoperatively to 54.2%,55.9%,59.0%,61.0% at 3,7,14,21 days postoperatively.The apoptosis indexes of hepatocells from group A to E were 8.1%,12.2%,19.4%,20.1%,14.2% respectively.Conclusions SPVE leads to atrophy of the ipsilateral hepatic lobe and hypertrophy of contralateral lobe,indicating that hepatocytes undergone apoptosis,rather than necrosis.The time point is 7 to 14 days.

OBJECTIVE: To explore the mutation of Y-STR loci in meiotic allelic transmission in a large pedigree. METHODS: The oral swabs of 163 male individuals were collected from a Lin pedigree. Twenty-two Y-STR genetic markers were typed with AGCU Y24 fluorescent detection kit (AGCU Y24 system), which also contained 16 Y-STR markers included in Yfiler multiple amplification kit (Yfiler system). The genotyping results of Y-STR loci were compared between each two males in the pedigree. RESULTS: There were 20 and 30 kinds of haplotypes obtained with Yfiler and AGCU Y24 systems in 163 male individuals from the Lin pedigree, respectively. The rates referred to haplotype differences (RRHD) of these two typing systems between male pairs were 0.910 5 and 0.922 7, respectively. The average number of marker differences were 6.582 1 and 9.824 8, respectively. The RRHD increased along with the incidents of meiosis. CONCLUSION Y-STR mutation leads to different Y-STR haplotypes among the male members in a paternal pedigree and the rate of difference increases along with the incidents of meiosis.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA Fingerprinting ; Genetic Linkage ; Genetic Markers/physiology* ; Genotype ; Haplotypes ; Humans ; Male ; Mutation/genetics* ; Pedigree

OBJECTIVE: To establish a 15-plex rapid STR multiplex amplification system. METHODS: Fourteen auto-chromosome loci and one sex-chromosome were selected to compare the situations of allelic losses and nonspecific amplication under different conditions. FastStart Taq DNA polymerase and DNA standard sample 9947A were used during amplification and optimization process.15-plex rapid STR amplification system was achieved by performing various experiments including selection of amplification conditions and the volume of DNA polymerase, adjustment of inter-locus balance, optimization of rapid amplification, screening of reaction buffers, selection of reaction volume, and a variety of additives. RESULTS: Using 10 μL rapid PCR system, including 1 ng DNA templates, 0.4 μL polymerase and 10xFastStart high fidelity reaction buffer, a complete and well-balance DNA profile of 15 STR loci for standard genomic DNA was obtained in 32 minutes, without the allele drop-out and non-specific amplicons. Meanwhile, 5% glycerinum, 0.01% gelatin, 0.05% gelatin and 5 mmol/L ammonium sulfate could be used as the reactive additive during the amplification procedure. CONCLUSION The 15-plex rapid STR multiplex amplification system can be used to decrease reaction time and enhance sample throughput.
Alleles ; Chromosome Mapping ; DNA/genetics* ; DNA Fingerprinting/methods* ; Forensic Genetics/methods* ; Humans ; Microsatellite Repeats/genetics* ; Polymerase Chain Reaction/methods* ; Racial Groups/genetics* ; Sensitivity and Specificity ; Tandem Repeat Sequences

0.05).Conclusion The distribution of G990R CASR genotype in PHPT patients is different from healthy women,and R allele is higher in PHPT group.Among PHPT patients,A986S and G990R polymorphisms are associated with serum calcium and ICa levels.Patients with S or G allele have lower levels of serum calcium and ICa.A986S genotype is also associated with serum PTH level and patients with S allele have relatively lower level of PTH.

BACKGROUNDEarly diagnosis and timely treatment are important for improving therapeutic efficiency of prostate cancer. DNA array is a new bio-technology for disease diagnosis. This study was conducted to diagnose prostate cancer with cDNA macroarray and analysis gene expression profiles of some selective genes in prostate cancer.

METHODSTotal RNA was isolated from patients with prostate cancer and from normal people, and poly (A) RNA was further purified. Then it was analyzed for differentially expressed genes in prostate cancer and normal prostate by cDNA macroarray system.

RESULTSThere were different expressions in the nine prostate-associated specific genes in prostate cancer as compared with normal prostate, in which, 7 were significantly upregulated and 2 were down-regulated.

CONCLUSIONAs a diagnostic approach at molecular level, the cDNA macroarray is an effectively diagnostic method for prostate cancer.

Gene Expression Profiling ; Genes, Tumor Suppressor ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis ; genetics

OBJECTIVETo study the significance of toll-like receptors (TLR) -7 and -8 in the pathogenesis of infection caused by Enterovirus type 71 (EV71) through measuring the expression of TLR7 and TLR8 in brain and lung tissues from the death cases caused by EV71 infection.

METHODSNine children who died of EV71 infection (EV71 group) were selected as study subjects, and 7 children who died of accidents or non-infectious diseases were used as the control group. Brain and lung tissues from the death cases in both groups at autopsy were collected, and immunohistochemistry was applied to detect the expression of TLR7 and TLR8 in lung and brain tissues in both groups. Integrated optical density (IOD) was applied for semi-quantitative analysis of the expression of TLR7 and TLR8.

RESULTSImmunohistochemical results showed that the expression of TLR7 and TLR8 in lung and brain tissues was strongly positive in the EV71 group, and the IOD values in the EV71 group were also significantly higher than those in the control group (P<0.05). There was no significant difference in the expression of TLR7 and TLR8 between lung and brain tissues in the EV71 group (P>0.05).

CONCLUSIONSTLR7 and TLR8 are highly expressed in lung and brain tissues from the patients who die of severe EV71 infection, suggesting that TLR7 and TLR8 may be involved in the pathogenesis of brain and lung damages caused by severe EV71 infection.

Brain ; immunology ; Child ; Cytokines ; physiology ; Enterovirus A, Human ; Enterovirus Infections ; etiology ; immunology ; Humans ; Lung ; immunology ; Toll-Like Receptor 7 ; analysis ; physiology ; Toll-Like Receptor 8 ; analysis ; physiology

OBJECTIVETo investigate and compare the short-term outcome of patients with low-middle frequency sudden deafness treated with alone or combination treatment.

METHODSFrom August 2007 to October 2011, 205 patients with the diagnosis of low-middle frequency sudden deafness who were from 33 different clinical centers were recruited. All patients were followed up for four weeks from the initial examination. Patients were treated with steroid , Ginaton, batroxobin respectively, or Ginaton and steroid combination treatment.

RESULTSThe total effective rate was 90.73%. In Ginaton group, the total effective rate was 87.27%, 89.19% in steroid group, 87.80% in batroxobin group, and 95.83% in Ginaton and steroid group. Considering the total effective rate, there was no statistical difference between four groups (χ(2) = 7.98, P = 0.54). The clinical cure rate for steroid alone was 81.01%, Ginaton alone 76.36%, batroxobin alone 68.29%, and Ginaton and steroid combination treatment 80.56%. There were no clinically significant differences between the different treatments (P > 0.05).

CONCLUSIONSThe low-middle frequency sudden deafness tends to have a relatively favorable prognosis. The steroid played a good effect in the treatment. But different treatments either improving the microcirculation of inner ear or alleviating edema blood has undifferentiated results. Therefore the combination therapy may be more effective.

Batroxobin ; China ; epidemiology ; Combined Modality Therapy ; Drug Therapy, Combination ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; classification ; diagnosis ; epidemiology ; therapy ; Humans

OBJECTIVETo identify the genetic cause for a Chinese Han family affected with hereditary multiple osteochondromas.

METHODSTwo patients, five unaffected relatives of the family and 100 unrelated healthy controls were collected. The coding sequences and intron/exon boundaries of EXT1 gene were amplified with polymerase chain reaction (PCR) and sequenced.

RESULTSA heterozygous c.600G>A (p.Trp200X) mutation in exon 1 of the EXT1 gene was detected in the patients. The same mutation was not found in unaffected family members and 100 healthy controls.

CONCLUSIONThe hereditary multiple osteochondromas in the family is caused by a nonsense mutation (p.Trp200X) in the EXT1 gene.

Asian Continental Ancestry Group ; genetics ; Child ; Exostoses, Multiple Hereditary ; diagnosis ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation ; N-Acetylglucosaminyltransferases ; genetics ; Pedigree