Objective This study aims to investigate the mutation spectrum and frequency of GJB2 , mtDNA12SrRNA,and SLC26A4 genes in Hui people,Tibetan,Tu nationality,and Mongolian patients with non-syndromic hearing loss in Qinghai province.Methods Peripheral blood samples were obtained from a total of 211 minority patients with nonsyndromic hearing loss in Qinghai province to extract genomic DNA.Three genes of GJB2,mitochondrialDNA12SrRNA,and SLC26A4 were screened for mutations in our study cohort using SNPscan technology.Results Among these 211 patients,5 Tu patients and 1 Mongolian patient were found to carry the ho-moplasmic mtDNAA1555G mutation.The GJB2 mutations detection rates were 11.38%,4.55%,5.88%,and 10%in Hui people,Tibetan,Tu nationality,and Mongolian patients,respectively.No statistically significant differences in the GJB2 mutations detection rates were found among all four ethnicities (P>0.05).c.235delC was the most prevalent mutation in both Tu patients and Mongolian patients.The allele frequency was 2.94% and 5%,respec-tively.While for Hui patients,c.299 300delAT was the most prevalent mutation with the allele frequency of 4.47%.The mutations detection rates of SLC26A4 were 6.5%,4.55%and 2.94%in Hui people,Tibetan,and Tu nationality patients,respectively.No statistically significant differences in the SLC26A4 mutations detection rates were found among all three ethnicities (P>0.05).c.235delC was the most prevalent mutation in Hui patients,the allele frequency was 2.44%.While for Tibetan patients,c.1226G>A was the most prevalent mutation with allele frequency of 2.27%.Conclusion A total of 10.9% of deaf patients have inherited hearing impairment caused by GJB2,SLC26A4,and mtDNAA1555G mutations.The mutation spectrum of GJB2 and SLC26A4 genes has the eth-nic specificity in nonsyndromic hearing loss patients of minority ethnicities in Qinghai province.

AIM: To construct a recombinant plasmid vector containing human pancreatic duodenal homeobox1(Pdx1) and neurogenic differentiation 1(NeuroD1) genes,and to detect its effective expression in eukaryocytes and the ability to induce differentiation of hepatocytes.METHODS: Using human embryo pancreas mRNA as template,Pdx1 and NeuroD1 genes were amplified by RT-PCR and cloned into the two different multiple cloning sites(MCSA and MCSB) of plasmid pIRES.The recombinant plasmid pI/Pdx1/NeuroD1 was transfected into L02 cells.The expression of Pdx1 and NeuroD1 in transfected cells was detected by immunocytochemistry,IFA,RT-PCR and Western blotting,respectively.RESULTS: The length and sequence of cloned segments were correct.Pdx1 and NeuroD1 were expressed in eukaryocytes.Furthermore,the hepatic cells were induced to express glucose transporter 2(GLUT2) and eukaryocyte transcription factor Nkx6.1,which were functionally correlated to ? cells.CONCLUSION: pI/Pdx1/NeuroD1 plasmid is successfully constructed and expressed in human eukaryocytes,with which the cells express the eukaryocyte transcription factor and GLUT2,indicating the transfected cells functionally correlates to ? cells.The results suggest that Pdx1 and NeuroD1 genes can induce the differentiation the cells from hepatic cells to pancreatic endocrine secretion cells.

AIM:To differentiate bone marrow mesenchymal stem cells(BMSCs) into functional insulin-producing cells to produce sufficient pancreatic islet cells for transplantation.METHODS:Recombinant adenovirus vectors carrying PDX1 and NKX6.1 genes were constructed and the bone marrow mesenchymal stem cells were infected by the recombinant adenovirus combined with several cytokines for differentiation.The expressions of PDX1,NKX6.1 and insulin and C-peptide in the differentiated bone marrow mesenchymal stem cells were detected by RT-PCR and Western blotting.After the differentiated bone marrow mesenchymal stem cells were transplanted into subrenal capsule of diabetic mice,cell morphology of the grafts as well as their secretion of insulin and C-peptide were detected.Besides,regulating capacities of grafts on the blood glucose level of the diabetic mice were also detected.RESULTS:BMSCs induced by recombinant adenovirus(pAdxsi-CMV-PDX1/CMV-NKX6.1) and several cytokines showed positive dithizone staining and the expressions of ?-cell related molecule such as insulin and glucose transporter 2 were detected by RT-PCR,which showed a sustaining and stable expression.The similar results were showed by Western blotting,immunohistochemical staining and indirect immunofluorescence.The insulin secretions in the cells stimulated with glucose at concentrations of 5.5 and 25 mmol/L in the experimental group were(1 240.4?109.3) mU/L and(3 539.8?245.1) mU/L,respectively,and were significantly higher than those in control group.Moreover,transplantation of the cells to STZ mice in treatment group made serum glucose recover to normal level.CONCLUSION:PDX1 and NKX6.1 gene-modified bone marrow mesenchymal stem cells differentiate into insulin-producing cells in vitro.When these cells transplanted into STZ induced diabetic mice,their serum glucose could return to the normal level and they could live well.Thus this is a promising method for diabetes treatment.

Objective To investigate the anatomical features, operative method and efficacy of internal fixation in the treatment of iutra-articular fractures of caleaneum via the sinus tarsi approach. Methods The pathway, branches distribution and anastomosis of perforating descending branch of peroneal artery were observed on 18 adult cadaveric lower limbs. A sinus tarsi approach was designed. From July 2001 to January 2008, 71 intra-articular calcaneal fractures in 68 patients were treated with open reduction and internal fixation via sinus tarsi approach at lateral sides of calcaneus. According to the Sanders classification, there were 26 type Ⅱ fractures, 32 type Ⅲ fractures and 13 type Ⅳ fractures. Results All patients were followed up for a mean period of 39.3 months (13-85 months), and the fractures were completely healed. There was a significant difference in the length, width and height of the calcaneus, Bohler angle and Gissane angle before and after operation (P < 0.01). According to Maryland Foot Score, the operative effect was excellent in 33 feet, good in 29 feet, fair in 6 feet and poor in 3 feet. Conclusion Open reduction and internal fixation via sinus tarsi approach is an effective method for minimally invasive treatment of intraarticular fractures of the calcaneus, with the advantages of good clinical results and causing minimal damage to soft tissues.

Objective To investigate the prognostic factors and survival of patients with bronchopulmonary carcinoid tumors (BPC) after surgical treatment .Methods The clinical data of 87 patients undergoing surgery for BPC from Jan .2002 to Dec .2008 were reviewed retrospectively .Kaplan-Meier method was used to analyze the survival of the patients .The risk factors such as age , gender ,smoking history ,histological type ,tumor size ,were analyzed by Cox proportional hazards regression model .Results The 1-,3-and 5-year overall survival rates were 85 .1% ,71 .3% and 63 .2% .Univariate analysis revealed that age (P=0 .016) ,smoking history(P=0 .007) ,histological type(P=0 .000) ,tumor stage(P= 0 .000) ,tumor size(P= 0 .006) lymph node metastasis(P=0 .000) ,surgery type (P= 0 .045) and postoperative chemotherapy (P= 0 .000) were prognostic factors .Multivariate analysis showed that histological type(P=0 .008) ,tumor stage(P=0 .000) ,lymph node metastasis(P=0 .033) were independent prognostic factor .Conclusion The survival rate of the BPC patient after surgical treatment is high ,histological type ,tumor stage and lymph node metastasis were independent prognostic factors .

Objective To investigate the influence of ursolic acid on vascular endothelial growth factor ( VEGF) , cycloxygen-ase-2 (COX-2), and matrix metalloproteinases-2 (MMP-2) expressed in the mouse retinal ischemic model , and to explore the mecha-nisms of anti-angiogenesis.Methods Sixty 7-day clean-class C57BL/6J mice were divided randomly into 6 groups [ n =10 mice (20 eyes) per group]:blank control, model control (PBS), positive control (triamcinolone), and ursolic acid (UA) intervention (low-dose, medium-dose, and high-dose).Mice in the blank control group were raised in air , and mice in other groups in(75%±2%)O2 high-oxygen environment for 5 consecutive days .Mice in the model control group and breastfeeding mice were put back in air environ-ment (21%O2 ) on the 12th day after the new-born mice to induce the generation of retinal neovascularization .When models were suc-cessful, the drug treatments were applied immediately to the corresponding groups , with injection of 3μl of sterile PBS in model control group, 3 μl of 1.5, 3.00 and 6.0 μg UA in UA intervention group, and 3 μl of triamcinolone (1 ml∶40 mg) in positive control group, respectively.All mice were killed after overdose anesthesia on the 17th day.Their eyeballs were made into samples and retinal tissue pathological sections with H-E dying method.The positive expressions of VEGF , COX-2, and MMP-2 were detected with immu-nohistochemical method .The fresh retinal tissue homogenate was prepared to detect the protein expressions of VEGF , COX-2, and MMP-2 in retinal tissue with western blot method ,and mRNA expressions of VEGF , COX-2, and MMP-2 were detected with real-time fluorescent quantitative polymerase chain reaction ( RT-PCR) .Results According to protein and mRNA expressions of VEGF , COX-2,and MMP-2 in retinal tissue among six groups , protein expressions of VEGF , COX-2, and MMP-2 in model group were significantly higher than those in blank group ( P <0.05 ) .Each protein expression in the high UA intervention group was significantly lower than that in the model group ( P <0.05 ) .Each protein expression in the high UA intervention group was not significantly different from that in the positive group ( P >0.05 ) .Each protein expression in the high UA intervention group was significantly lower than that in the low UA intervention group( P <0.05).Conclusions UA inhibited expressions of VEGF, COX-2, and MMP-2 in retinal ischemia model .UA also played an inhibitory role in the formation of neovascularization , and this role was positively correlated with UA dose .

Comprehensive treatment dominated by surgery is the mainstay in the treatment of hepatic cancer,and hepatectomy is still the most effective treatment method.Bile duct reconstruction after hepatectomy is still the difficult point for the treatment of hepatic cancer complicated by bile duct invasion.A 45-year-old patient with hepatic cancer and gallstone was admitted to the Affiliated Hospital of Guiyang Medical College,magnetic resonance cholangiopancreatography and enhanced computed tomography indicated that intrahepatic duct was dilated and tumor had invaded both left and right hepatic ducts.Cholecystectomy,mesohepatectomy,duct to duct anastomosis of left hepatic duct and common hepatic duct,duct to duct anastomosis of right hepatic duct and cystic duct were performed during the operation.The patient was cured 2 weeks after surgery.

OBJECTIVETo investigate the changes in the quantity of colonizing Streptococcus mutans(S. mutans) and Actinomyces on the root surface plaque before and after post-core crown restoration of the mandibular first molars in the elderly patients.

METHODSA total of 30 elderly patients, each with one post-core crown restoration of the mandibular first molar, were randomly chosen to participate in the studies. Patients with mandibular first molars with post-core crown restoration and those with healthy contralateral mandibular first molars were divided into the test and control groups, respectively. Root surface plaques of the two groups were collected before tooth preparation, 72 h after preparation, one week after preparation, and one month after restoration. S. mutans, Actinomyces naeslundii (A. naeslundii) and Actinomyces viscosus (A. viscosus), were identified using colony morphology, biochemical techniques, and polymerase chain reaction (PCR). Plaque count was measured using microbial colony count.

RESULTSThe number of S. mutans and A. viscosus and A. naeslundii in the test group, which was statistically significant (P<0.05), increased 72 h after preparation. The quantities of S. mutans, A. viscosus, and A. naeslundii one week after preparation were significantly different (P<0.05). The plaque count of S. mutans, A. viscosus, and A. naeslundii in the test group decreased one month after restoration (P<0.05).

CONCLUSIONThe quantities of S. mutans, A. viscosus and A. naeslundii increase one week after preparation but decrease one month after restoration. The finding suggests that dentists should educate patients about plaque control during the early period after tooth preparation.

Actinomyces ; Actinomyces viscosus ; Aged ; Bacteria ; Crowns ; Dental Plaque ; Humans ; Post and Core Technique ; Streptococcus mutans ; Tooth Root

Objective To describe the US, CT, and MR imaging findings and diagnosis of hepatic adenomas. Methods The comprehensive imaging features in 6 patients with 6 hepatic adenomas confirmed pathologically were reviewed retrospectively and correlated with pathologic findings. Results One case was diagnosed correctly, four cases were mistaken for hepatocellular carcinomas (HCC), and one case was mistaken for focal nodular hyperplasia. US: six lesions were hypoechoic with hypohalo in four lesions, and there was low velocity arterial and venous flow within the six lesions. CT: six lesions were hypodense with pseudocapsule in four lesions, and the four lesions showed slight enhancement during arterial and portal venous phases, and one lesion showed moderate enhancement during arterial phase and slight enhancement during portal venous phase. MRI: six lesions had heterogeneously high signal intensity on T 1WI and T 2WI, and the high intensity on T 1WI remained unchanged after using fat saturation. Two lesions showed strong enhancement during arterial phase and slight enhancement during portal venous and delayed phases, and three lesions showed slight enhancement during arterial, portal venous, and delayed phases. Pseudocapsule detected in six lesions showed slight enhancement on portal venous or delayed phases. Conclusion The comprehensive imaging findings of hepatic adenomas were nonspecific. The presence of pseudocapsule, heterogeneous high signal intensity on T 1WI, and the high intensity remained unchanged after using fat saturation may help make a correct diagnosis of hepatic adenoma.

An analysis of the opportunities and challenges brought forth to research hospitals in the face of emerging new technologies,described the development direction of such hospitals in terms of strategic planning of precision medicine, breakthrough of mechanism barriers for technology development,andInternet+ service mode innovation.These efforts aim at exploring to build a new type of medical technology service system.