A novel member of extremely halophilic archaea, strain AJ2, was isolated from Ayakekum Lake located in Altun Mountain National Nature Reserve of Xinjiang Uygur Autonomous Region in China. The strain AJ2 requires at least 10% (w/v) NaCl and grows 10% to 30% (optimum at 20%). Phylogenetic analysis based on 16S rDNA sequence comparison revealed that strain AJ2 clustered to three Natrinema species with less than 97.7% sequence similarities, suggesting AJ2 is a novel member of Natrinema. A bacteriorhodopsin-encoding (bop) gene was subsequently detected in the AJ2 genome using the polymerase chain reaction technique. The cloning and sequencing of a 401 base pairs fragment indicated the deduced amino acid sequence of bop from AJ2 is different from that reported for bacteriorhodopsins. This is the first reported detection of a bop gene in Natrinema.
Bacteriorhodopsins ; genetics ; metabolism ; Cell Proliferation ; Gene Expression Profiling ; Genome, Archaeal ; Halobacteriaceae ; classification ; isolation & purification ; physiology ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity

A novel member of extremely halophilic archaea, strain AJ2, was isolated from Ayakekum Lake located in Altun Mountain National Nature Reserve of Xinjiang Uygur Autonomous Region in China. The strain A J2 requires at least 10% (w/v)NaCl and grows 10% to 30% (optimum at 20%). Phylogenetic analysis based on 16S rDNA sequence comparison revealed that strain A J2 clustered to three Natrinema species with less than 97.7% sequence similarities, suggesting A J2 is a novel member of Natrinema. A bacteriorhodopsin-encoding (bop) gene was subsequently detected in the A J2 genome using the polymerase chain reaction technique. The cloning and sequencing of a 401 base pairs fragment indicated the deduced amino acid sequence of bop from A J2 is different from that reported for bacteriorhodopsins. This is the first reported detection of a bop gene in Natrinema.

Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.

OBJECTIVE: To evaluate the VMA/Cr and HVA/Cr of random urine in the diagnosis of neuroblastoma. METHODS: VMA/Cr and HVA/Cr were examined in 50 normal children and 11 cases of neuroblastoma (NB) aged 1 approximate, equals 5 years. The concentration of 4-hydroxy-3-methoxy-mandelic acid (VMA) and homovanillic acid (HVA) in random urine collection were analyzed by high-performance liquid chromatography and creatinine (Cr) analyzed by auto-biochemical analyzer. RESULTS: The normal range of VMA/Cr and HVA/Cr were <14.9 mmol/mol and <23.8 mmol/mol, respectively. The positive rate of VMA/Cr and HVA/Cr in NB patients was 90.9%. CONCLUSION: Determination of VMA/Cr and HVA/Cr in random urine collection can be used for NB diagnosis.

Objective To report a three-generation Chinese family with freckle and to make a genetic linkage analysis in this family.MethodsGenetic linkage analysis was carried out in this family using microsatellite markers distributed over chromosome 4q and 1.Two-point logarithm of odds(LOD)scores were calculated using the Linkage program package(version 5.1),and haplotype was analyzed with Cyrillic version 2.01 software.Results Freckle was inherited in an autosomal dominant pattern with a penetrance of99.9% in this family;linkage to chromosome 4q was ruled out however,supportive evidence was obtained for linkage to microsatellite markers D1S2635 and D1S2844 in chromosome 1q with a maximum LOD score of 1.50.Haplotype analysis in this family localized the locus of freckle to a 12 Mb region flanked by D1S2624 and D1S2799.Conclusions Freckle is a genetically heterogeneous disorder.The causative gene may be located in a 21.2 cM region on chromosome 1q22-24.

Objective： A prospective randomized trial was performed to evaluate the contribution of neoadjuvant chemotherapy in the patients with locoregionally advanced nasopharyngeal carcinoma. Methods： The patients with locoregionally advanced nasopharyngeal carcinoma were treated with either radiotherapy alone (RT group) or neoadjuvant chemotherapy plus radiotherapy (CT/RT group). Neoadjuvant chemotherapy consisting of 2-3 cycles of cisplatin (20 mg/m2 on Day 1 to Day 5), bleomycin (7 mg/m2 on Day 1 and Day 5), and 5-FU (500 mg/m2 on Day 1 to Day 5) followed by radiotherapy was given to CT/RT group. All patients were treated in a uniform definitive-intent radiation therapy in two groups. Results： From 1992 to 1993, 457 patients were enrolled and 440 patients (221 in RT group, 219 in CT/RT group) were assessable. The 5-year overall survival (OS) rates were 62％ for CT/RT group and 55％ for RT group (P=0.1335)； The 5-year relapse free survival rate was 48％ versus 58％ , respectively (P=0.0539). The 5-year free local recurrence (FLR) rate was 82％ for CT/RT group, 74％ for RT group (P=0.0412). There was no significant difference in free distant metastasis (FDM) between two treatment groups (CT/RT group, 79％ ; RT group, 75％ ; P=0.4177). Subgroup analyses showed that neoadjuvant chemotherapy improved local control in patients with T3-4 disease, and had no effect in preventing distant metastases in patients with N2-3 disease. Conclusion： Despite improving FLR and RFS, neoadjuvant chemotherapy and radiatherapy failed to gain other survival benefit or reduce distant metastases in patients with locoregionally advanced nasopharyngeal carcinoma. The indication for neoadjuvant chemotherapy is proposed.

Objective:To explore correlation between fibrinogen(Fg)level and coronary stenotic degree in aged coronary heart disease(CHD)people and its possible pathogenesis.Methods:A total of 112 patients,who were >65 years and diag-nosed as CHD by coronary angiography(CAG),were selected as CHD group.Another 108 non-CHD patients hospitalized during the same period were enrolled as non-CHD group.According to CAG results,CHD group was divided into single vessel coronary disease group(n=30,single vessel group)and multi vessel coronary disease group(n= 82,multi vessel group).Serum levels of Fg,D-dimer(D-D),C reactive protein(CRP),homocysteine(Hcy)and brain natriuretic peptide (BNP)were measured and compared between two groups.The correlation between serum Fg level and Gensini score was analyzed in CHD patients.Results:Compared with non-CHD group,there were significant rise in serum levels of Fg,D-D, triglyceride(TG),apolipoprotein B(ApoB),lipoprotein a,CRP,BNP and Hcy in CHD group,P=0.001 all;in CHD group,compared with single vessel group,there were significant rise in serum levels of Fg,D-D,ApoB,uric acid,lipo-protein a,CRP,BNP,Hcy,and significant reductions in serum levels of high density lipoprotein cholesterol,ApoA1 and ApoA1/ApoB in multi vessel group,P<0.05 or <0.01. Serum Fg level was significant positively correlated with coronary stenotic degree(Gensini score),r=0.303,P=0.001. Fg was independent risk factor of CHD(OR=2.74,95% CI:1.95~3.97,P=0.001).Conclusion:Serum Fg level is significant positively correlated with coronary stenotic degree in aged CHD patients.Its mechanism of triggering atherosclerosis may be related to risk factors such as dyslipidemia,hyper-homocysteinemia and inflammations etc.

OBJECTIVETo investigate the frequency of micrometastasis in levels lII - IV of clinical negative neck (cN0) in patients with squamous cell carcinoma (SCC) of oral tongue, and to discuss the management of cervical lymph node for cN0 tongue SCC.

METHODSA total of 471 cervical lymph nodes derived from 25 patients with cN0 tongue SCC, including 263 lymph nodes in level III and 208 lymph nodes in level IV, were included in this study. All lymph nodes were re-examined by anti-cytokeratin (CK) immunohistochemical staining combined with semi-serial section per 500 microm.

RESULTSAmong the 25 cases, seven patients were confirmed harboring metastasis in 11 lymph nodes of level III, and no positive lymph node in level IV was detected by routine hematoxylin-eosin (HE) staining. 11 positive lymph nodes in level IIl, which confirmed by HE staining, were also detected by immunohistochemical staining with CK combined with semiserial section. Among the 460 cervical lymph nodes in which HE staining did not show metastasis, only one lymph node in level III harboring a 2.0 mm x 1.5 mm micrometastasis was detected by immunohistochemical staining with CK, and no positive lymph node in level IV was detected by immunohistochemical staining with CK.

CONCLUSIONThe frequency of occult metastasis in level IV was very low, so it seemed unnecessary to dissect level IV for all patients with cN0 tongue SCC.

Adult ; Aged ; Carcinoma, Squamous Cell ; Female ; Humans ; Keratins ; Lymph Nodes ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck ; Neoplasm Micrometastasis ; Tongue ; Tongue Neoplasms

BACKGROUNDMolecular hydrogen, as a novel antioxidant, has been proven effective in treating many diseases. This study aimed to evaluate the therapeutic effects of hydrogen saturated saline in treatment of a rat model of diabetes mellitus and a rat model of insulin resistant.

METHODSA rat diabetes mellitus model was established by feeding a high fat/high carbohydrate diet followed by injection of a small dose of streptozotocin, and an insulin resistant model was induced with a high glucose and high fat diet. Hydrogen saturated saline was administered to rats with both models conditions on a daily basis for eight weeks. A pioglitazone-treated group and normal saline-treated group served as positive and negative controls. The general condition, body weight, blood glucose, blood lipids, and serum insulin levels of rats were examined at the 8th week after treatment. The oxidative stress indices, including serum superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) were also evaluated after eight weeks of treatment using the commercial kits.

RESULTSHydrogen saturated saline showed great efficiency in improving the insulin sensitivity and lowering blood glucose and lipids. Meanwhile, the therapeutic effects of hydrogen saturated saline were superior to those of pioglitazone. Hydrogen saturated saline markedly attenuated the MDA level and elevated the levels of antioxidants SOD and GSH.

CONCLUSIONHydrogen saturated saline may improve the insulin resistance and alleviate the symptoms of diabetes mellitus by reducing the oxidative stress and enhancing the anti-oxidant system.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; Hydrogen ; therapeutic use ; Hypoglycemic Agents ; therapeutic use ; Insulin Resistance ; Oxidative Stress ; drug effects ; Rats ; Sodium Chloride ; chemistry ; Thiazolidinediones ; therapeutic use

OBJECTIVETo evaluate the biocompatibility of pectin/poly vinyl alcohol composite (CoPP) hydrogel for use as a prosthetic nucleus pulposus material.

METHODSThe in vitro cytotoxicity of CoPP hydrogel was tested in NCTC L929 cells, which were divided into normal control group, negative control group [treated with poly (vinyl alcohol) hydrogel, PVA], experimental group (treated with CoPP) and positive control group (0.64% phenol). The optical density of the cells on days 2, 4, and 7 of the corresponding treatments was determined and the relative growth rate calculated. For in vivo biocompatibility evaluation, dehydrated CoPP and PVA hydrogel were respectively implanted into the left and right gluteus of SD rats, and the wound healing and general status were observed. The muscular tissues containing the implants were taken 1, 4, and 12 weeks after the implantation for gross observation and microscopic observation of the inflammatory cell infiltration (ICI) and formation of the fibrous capsulation around the implants.

RESULTSThe L929 cells incubated with PVA and CoPP group both grew well, with relative growth rate over 80% and 75%, respectively. The cytotoxicity of PVA and CoPP was both lower than grade 1. In contrast, the relative growth rate in the positive control group was below 24%, with cytotoxicity over grade 4. In the SD rats, ICI of grade IV occurred in the muscular tissues around the PVA and CoPP implants at 1 week without formation of complete capsule, and at 4 weeks, ICI was lowered to grade 1 with grade 4 capsular reaction. Till week 12, the ICI and capsular reaction were both first grade.

CONCLUSIONCoPP hydrogel has in vitro grade 0 or 1 cytotoxicity and causes only mild inflammation after implantation in rats, suggesting good biocompatibility of the material.

Animals ; Biocompatible Materials ; Hydrogel, Polyethylene Glycol Dimethacrylate ; Implants, Experimental ; Intervertebral Disc ; surgery ; Materials Testing ; methods ; Pectins ; chemistry ; Polyvinyl Alcohol ; chemistry ; Rats