OBJECTIVETo investigate the whole sequence of SLC26A4 gene among 1552 deaf students from 21 regions of China with SLC26A4 hot spot mutation IVS7-2A > G and analyze the epidemiological state of enlarged-vestibular-aqueduct-syndrome (EVAS) related hearing loss in China.

METHODSDNA was extracted from peripheral blood of 1552 students from deaf and dumb school of 21 regions in China. The nationality of the 1552 cases covers Han (1290 cases), Uigur (69 cases), Hui (37 cases), Mongolia (31 cases), Yi, Zhuang, Bai, Miao and other 13 nationalities (125 cases). Firstly, all subjects were analyzed for the hot spot mutation IVS7-2A > G by direct sequencing. Those carrying a single heterozygous IVS7-2A > G were given further analyzed for the probable second mutation in other exons except exon7 and exon8 of SLC26A4. One hundred and fifty cases with normal hearing were in the control group.

RESULTSThe sequencing results revealed 197 cases carrying IVS7-2A > G, of whom 83 carrying IVS7-2A > G homozygous mutation, 114 carrying IVS7-2A > G heterozygous mutation. Of the 114 cases with heterozygous IVS7-2A > G, 78 cases were found to have another mutation and 36 cases were found no other mutation in SLC26A4. Of the 1552 cases, the percentage of cases carrying homozygous IVS7-2A > G and compound heterozygous mutations was 10.37% (161/1552). Of the 78 cases with SLC26A4 compound heterozygous mutations, the mutations except IVS7-2A > G were found mainly in exon 19, 10, 17, 15, 11 + 12, 14 and 3. Twenty-one novel SLC26A4 mutations were found. In the control group, there were only 3 cases carrying heterozygous IVS7-2A > G, and no other mutation in SLC26A4 was found.

CONCLUSIONSSLC26A4 mutations account for at least 10% of EVAS related hereditary hearing loss in China. It's of great importance to screen SLC26A4 gene for making aetiological diagnosis for deafness. The discovery of novel variants of SLC26A4 gene makes the mutational and polymorphic spectrum more plentiful in Chinese population. We also provide preliminary evidence for the hot spot areas of SLC26A4.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; China ; epidemiology ; Female ; Hearing Loss, Sensorineural ; epidemiology ; ethnology ; genetics ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Sequence Analysis, DNA ; Vestibular Aqueduct

Glycoproteins ; genetics ; metabolism ; Herpesviridae ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo investigate the feasibility, effectiveness and practicability of transurethral enucleation plus pneumocystostomy rotary cut (TUE + PCRC) for large benign prostatic hyperplasia (BPH).

METHODSWe performed TUE + PCRC for 26 BPH patients aged 62 - 85 years with the prostate volume of 80 - 165 ml. We conducted transurethral enucleation of the hyperplastic prostate glands and pushed them into the bladder, followed by bladder puncture for pneumo-cystostomy rotary cut.

RESULTSAll the surgical procedures were successfully accomplished, with the mean surgical time of 41 (32 - 54) minutes and intraoperative blood loss < 60 ml in all the cases. Twenty-three of the patients were followed up for 2 - 8 months, which revealed no stricture of the urethra or any other severe complications. Compared with the preoperative baseline, significant improvement was achieved in the IPSS (6.5 +/- 2.2 vs 26.2 +/- 2.4), QOL (1.4 +/- 0.9 vs 4.6 +/- 1.2) and Qmax ([5.8 +/- 1.0 ] vs [19.6 +/- 2.8] ml/s) of the patients after surgery (P < 0.01).

CONCLUSIONTUE + PCRC, with its advantages of short operation time and less severe complications, is a safe and effective approach to the management of large BPH.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; surgery ; Transurethral Resection of Prostate ; methods

OBJECTIVETo summarize the clinical experience on the arterial switch operation for complex congenital heart disease in recent 3 years in our hospital.

METHODSSixty-one patients with complex congenital heart disease received by arterial switch operation from Jan 2000 to May 2003. There were 26 patients with transposition of the great arteries and intact ventricular septum (TGA-IVS), 19 patients with transposition of the great arteries and ventricular septal defect (TGA-VSD) and 16 patients with double outlet of right ventricle with subpulmonary ventricular septum defect (Taussig-Bing).

RESULTSThere were 2 death among TGA-IVS cases, 4 deaths among TGA-VSD cases and 4 deaths among Taussig-Bing cases. The total operative mortality was 16%. All patients were followed up 3 months to 3 years. In all patients, the cyanosis disappeared and the physical activities increased obviously. One patients with Taussig-Bing keep moderate mitral valve reflux unchanged, 2 patients with TGA had pulmonary and aortic supra-valve stenosis with 40 mmHg pressure gradient. One patient with subpulmonary stenosis and residual VSD was re-operated 3 months later. All survivor had good heart function.

CONCLUSIONSThe effect of arterial switch operation on the treatment of TGA was well accepted in this study. The procedure used in treatment on Taussig-Bing can prevent pulmonary obstruction and avoid the complication of left ventricular outflow tract obstruction caused by intraventricular repair.

Aorta ; surgery ; Cardiovascular Surgical Procedures ; methods ; Double Outlet Right Ventricle ; complications ; surgery ; Female ; Follow-Up Studies ; Heart Septal Defects, Ventricular ; complications ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Pulmonary Artery ; surgery ; Retrospective Studies ; Transposition of Great Vessels ; complications ; surgery

OBJECTIVETo investigate the etiology of patients with severe to profound hearing loss and to identify the ratio of hereditary hearing loss in Chifeng area in Northern China.

METHODSDNA were extracted from peripheral blood of 134 deaf patients from Chifeng special educational school and 100 normal hearing controls in Northern China. Audiology examinations showed that all patients had severe to profound bilateral sensorineural hearing impairment. Sequence analysis of the whole coding areas of GJB2, GJB3, GJB6, SLC26A4, mtDNA12SrRNA and mtDNAtRNASer(UCN) were performed. Individuals carrying SLC26A4 mutation were given further temporal bone CT scan.

RESULTSThe ratio of hearing loss related to genetic factors in this population was 60.45% (81/134). About 33.58% (45/134) of the patients were given accurate genetic diagnosis. GJB2 mutations were responsible for approximately 17.16% of the cases in ChiFeng area. By screening SLC26A4 followed by temporal bone CT scan, we diagnosed 20 cases of enlarged vestibular aqueduct (EVA) and/or other inner ear malformation. SLC26A4 mutations account for about 14.93% of the cases. The aminoglycoside-related mtDNA 1555A>G mutation accounted for 0.76% of the cases in Chifeng area. In addition, another 13.43% (18/134) of the cases carried heterozygous GJB2 mutation and their hearing loss may be related to GJB2. 6.72% (9/134) of the cases carried heterozygous SLC26A4 mutation who were not found EVA by temporal bone CT or not took CT examination for some reasons. However, their hearing loss may also be SLC26A4-related. About 2.24% (3/134) of the cases carried mtDNA 12SrRNA 1095 T>C which may also be an aminoglycoside-related mutation and very likely be the cause of hearing loss. GJB3 might participate in the pathomechanism of hearing loss in 1.49% (2/134) of the patients. GJB6 mutation was not detected in this population.

CONCLUSIONSThe ratio of hearing loss related to genetic factors in the sample drawing population from Chifeng was 60.45% (81 cases). GJB2 is the most common gene and SLC26A4 is the second common gene next to GJB2 that cause deafness in this area.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Connexin 26 ; Connexin 30 ; Connexins ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Gene Frequency ; Genotype ; Hearing Loss ; epidemiology ; genetics ; Heterozygote ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Young Adult

OBJECTIVETo investigate the effects of clinical and pathomorphological parameters on the prognosis of colon carcinoma and rectal carcinoma.

METHODSUnivariate and multivariate COX proportional hazard models were used to study the effects of the clinical and pathomorphological factors on the prognosis in 101 cases of colon carcinoma, 219 of rectal carcinoma and 137 of rectal carcinoma under curative resections.

RESULTBy using univariate analysis, we identified that lymph node metastasis and distant metastasis were the common prognostic factors for both colon carcinoma and rectal carcinoma. Smoking, deep infiltration, chemotherapy and serum albumin concentration were the uncertain prognostic factors for colon carcinoma. Signet-ring cell carcinoma, larger tumor size (>6 cm), deep infiltration, lack of radical surgery, and advanced TNM stage were the exclusive adverse prognostic factors for rectal carcinoma. Further studies showed that the adverse prognostic factors for the rectal carcinoma under curative resection included deep infiltration, lymph node metastasis, vessel invasion, less of peritumoral lymphocyte infiltration, lack of Crohn's like reactivity, high level of tumor budding, advanced TNM stage and positive urine glucose. By using multivariate analysis based on a COX proportional hazard model, it was identified that smoking, lymph node metastasis and serum albumin concentration were independent prognostic factors for colon carcinoma; advanced TNM stage, distant metastasis and palliative surgery for rectal carcinoma; and vessel invasion, lymph node metastasis and urine glucose for rectal carcinoma under curative resections.

CONCLUSIONThe various clinical and pathomorphological parameters show different prognostic value for colon carcinoma, rectal carcinoma and rectal carcinoma under curative resections.

Adult ; Aged ; Carcinoma, Signet Ring Cell ; pathology ; surgery ; Colonic Neoplasms ; pathology ; surgery ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Rectal Neoplasms ; pathology ; surgery

The tumor suppressor gene p53 and p16, both of which play an important role in inhibition of tumorigenesis, are homozygously deleted in human myeloid leukemia cell line K562. To explore the inhibition of K562 cell proliferation by wild type p16 and p53 genes, both p16 and p53 genes were co-transfected into K562 cells mediated by liposome. The expression of the two genes was measured by immunocytochemical method, the cell cycle was analysed by flow cytometry, and the number of recovered viable cells was assessed after transfection. After co-transfection, the p53 and p16 positive cells were 23% and 28%, respectively. The results showed that co-transfection of p16 and p53 genes significantly inhibits cell proliferation comparing with transfection either by p16 gene or by p53 gene (P < 0.05). Expression of p16 and p53 proteins increased the cell number in G(1) phase but decreased the cell number in S phase. It is concluded that co-transfection of p16 and p53 genes has a stronger growth-inhibitory effect on K562 cell growth than that of transfection only by p16 gene or by p53 gene, may be a pathway for gene therapy in leukemia.
Cell Division ; Genes, p16 ; physiology ; Genes, p53 ; physiology ; Humans ; K562 Cells ; Plasmids ; Transfection

A pair primer was designed by Oligo 6.0 according to the pilA gene sequence of E. coli isolated from human in GenBank. The pilA Gene was obtained by PCR with the enteropathogenic E. coli isolated from ducks as template and cloned into pMD18-T vector. It was identified by PCR, restriction endonuclease analysis, DNA sequencing and then subcloned into BamH I/Hind III site of prokaryotic expression vector pET-32a(+) and recombinant expression plasmid pET-32a-pilA was constructed successfully. The plasmid was transformed into Eschericha coli BL21 (DE3) and 36kD pilA recombinant protein was expressed be induced with IPTG. The protein was purified by Ni-agarose affinity chromatograghy and was prepared as vaccine with Freund' s adjuvant. The ducklings were immunized with the vaccine at 1 and 8-day-old respectively. Two weeks after last immunized, the antibody titer of duck serum was detected by ELISA and the ducklings were challenged with 10(9) PFU enteropathogenic E. coli GH1.2 virulent strain. The immunoprotection effect of pilA recombinant protein vaccine was evaluated according to the mortality, re-isolated rate of E. coli, and grades of pathological changes. The results show that the antibody titer are 1:12800, but 1:200 were detected from ducklings immunized with homologous whole cells E. coli inactivated vaccine. The mortality, re-isolated rate of E. coli, degree of pathological changes of immunized ducklings is lower than that of the control ducklings and showed significant or extremely significant differences (P < 0.01 or P < 0.05), but non-significant difference compared to the ducklings which immunized with homologous whole cells E. coli inactivated vaccine (P > 0.05). The results show that pilA recombinant protein has some immunoprotection effect with the challenging of virulent strains of E. coli GH1.2.
Animals ; Animals, Newborn ; Antibodies, Bacterial ; blood ; immunology ; Ducks ; microbiology ; Electrophoresis, Polyacrylamide Gel ; Enteropathogenic Escherichia coli ; genetics ; immunology ; pathogenicity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli Proteins ; genetics ; immunology ; metabolism ; Fimbriae Proteins ; genetics ; immunology ; metabolism ; Humans ; Immunization ; Polymerase Chain Reaction ; Poultry Diseases ; immunology ; mortality ; prevention & control ; Recombinant Proteins ; administration & dosage ; immunology ; metabolism ; Survival Rate ; Virulence

OBJECTIVETo study the influence of clinical and pathological-morphological parameters on the prognosis of colorectal carcinoma.

METHODSUnivariate and multivariate Cox proportional hazard model were used to study the influence of clinical and pathological-morphological factors on the prognosis in 226 colorectal carcinoma cases.

RESULTSUsing univariate analysis, data showed that the factors significantly related to disease prognosis would include: the depth of direct spread, vessel invasion, perineural invasion, tumor budding, peritumoral-lymphocytic infiltration, Crohn-like reaction, number of positive lymph nodes, distant metastasis, TNM stage and urine glucose. Multivariate Cox proportional hazard model showed that six factors were identified to be associated with higher relative-risk (RR), including: older age, advanced TNM stage, more severe budding, perineural invasion, less peritumoral-lymphocytic infiltration and urine glucose.

CONCLUSIONAge, TNM stage, tumor budding, perineural invasive, peritumoral-lymphocytic infiltration and urine glucose were independent predictors to the prognosis of colorectal carcinoma.

Aged ; China ; epidemiology ; Colorectal Neoplasms ; diagnosis ; epidemiology ; pathology ; Female ; Glycosuria ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Regression Analysis ; Retrospective Studies ; Risk Factors

Replication of duck plague virus(DPV) in artificially infected ducks were detected by in situ hybridization (ISH) which employed a 37bp oligonucleotide as probe designed according to DPV DNA sequence in GenBank. The results indicated that DPV DNA was detected in liver, intestine and bursa Fabricius at 4 h, in spleen and esophagus at 6h, in thymus at 12h post infection; DPV DNA in lung and kidney was detected only in dead ducks and no positive signal was detected in muscle, heart, cerebrum and pancreas. DPV DNA was distributed in cell nucleus and cytoplasm. Hepatocytes, sinus endodermal cells and Kuffer's cells were the mainly infected cell types in liver. DPV DNA was mainly detected in epithelium of villi, in lamina propria of intestinal villi of duodenum, in stratum spinosum of esophagus, and in epithelium, cortex, medulla of bursa Fabricius. The positive signals were mainly detected in medulla of thymus, lymphocytes and macrophages of spleen. The research suggests that ISH is a direct and specific method in detecting DPV DNA in paraffin sections and it's also a good method for virus diagnosis and DNA location of DPV.
Animals ; DNA, Viral ; analysis ; Ducks ; virology ; In Situ Hybridization ; Influenza A virus ; genetics ; isolation & purification ; physiology ; Virus Replication