Objective To study the technique of transposon mutagenesis involving the use of Mini-Tn5 on a suicide plasmid for Citrobacter freundii (C. freundii). Methods A conjugation method was used in transposon mutagenesis. The stability of exconjugants was confirmed by series passages under nonselective condition. Detection of anti-ampicillin resistance and purification of the plasmid were conducted for excluding the possibility of plasmid-encoded antibiotic resistance. All exconjugants were screened for auxotrophic mutants and swarming motility mutants. Results Analysis of the generated exconjugants indicated that Mini-Tn5 Km was transposed in C. freundii at a high frequency (ca. 9.2?10 -6). The mutants were stable and retained the transposon-encoded antibiotic resistance after incubation for three passages under nonselective condition. The frequency of auxotrophic mutants in the population confirmed that the insertion of the transposon was random and the C. freundii chromosome did not contain significant insertional hot spots of transposition. No extrachromosomal plasmid DNA was found. Conclusion Transposon mutagenesis of Mini-Tn5 is a suitable technique for the study of differentiation in C. freundii.

Objective To study variations of plasma sE-selectin and sICAM-1 level of type 2 diabetes(T2DM) patients complicated with lower limb vascular disease,and contribution of these molecules to the formation of the lower limb vascular disease. Methods Plasma sE-selectin and sICAM-1 of 51 type 2 diabetic patients and 20 age & sex matched non-diabetes people(control group) were determined with ELISA.16 T2DM patients without lower limb vascular disease and 17 age & sex matched T2DM complicated with lower limb vascular disease were selected from the patients group,as group 1 and group 2 respectively,and 17 age and sex matched non-diabetic individuals were selected from control group as group 3.sE-selectin and sICAM-1 level were contrasted among the three groups. Results Levels of FPG,2 h PG,HOMA-IR,sE-selectin and sICAM-1 of diabetic group were significantly elevated than those of the control group(P0.05).FPG and 2 h PG in group 1 and group 2 were markedly higher than that in group 3(P

Objective To comprehend the status of Helicobacter pylori (Hp) infection in type 2 diabetes mellitus (T2DM) patients, and study the relationship between Hp and T2DM. Methods Seven hundred and sixty-eight in-hospital T2DM patients (T2DM group, including new diagnosed 194 patients and re-diagnosed 574 patients) and 172 sex, age matched health individuals(control group ) were involved in this study. 14C-urea breath test (14C-UBT) was proceeded in both groups to determine Hp infection, and infection rates were calculated. Results Total Hp infection rate in T2DM group was significantly higher than that in control group [56.0%(430/768) vs 37.8%(65/172) ,P< 0.01], delamination analysis according to sex also showed significant differences between T2DM group and control group(P < 0.01). The rate of Hp infection in new diagnosed patients and re-diagnosed patients, female patients and male patients had no significant difference (P > 0.05 ). The rate of Hp infection in different age and sex had no significant difference (P >0.05 ). Conclusions The rate of Hp infection in T2DM patients is higher than that in non-T2DM patients.Hp infection has relationship with T2DM, and has no relationship with age, sex and course of disease.

A strain of P. Aeruginosa,which was seperated from clinical environment ,shows a special characteristic . It keeps normal short rod shape when cultured at 37℃,however,it forms filament without pyocyanin producing when cultured at 25℃ overnight. The filaments will divide and form short rods,simultaneously,produce pyocyanin when culture time is prolonged to over 72h or culture temperature is raised to 37℃. The preliminary study indicates that this phenomenin has nothing to do with nutritive conditions and could the inbluenced by inoculating density and irradiating with ultraviolet rays The absence of pyocyanin was not the cause of filamentous formation by the test results.

Objective To explore the technique and clinical outcome of the atlantoaxial pedicle screw system in the treatment of the unstable atlantoaxial dislocation post traction.Methods The study involved 32 patients with atlantoaxial dislocation(type T2 of TOI classification)admitted from March 2005 to September 2009.There were 17 patients with fresh odontoid fracture and two with old odontoid fracture,five with traumatic disruption of the transverse atlantal ligament and eight with congenital odontoid dysplasia.JOA scores of neurological function before operation was at a range of 5-13(average 8.38).The average of space available for the cord(SAC)was 9.15 mm.Before the atlantoaxial pedicle screw system was carried out,the skull traction was performed in all the patients preoperatively.Results A total of 128 pedicle screws were inserted safely,with mean operation time and perioperative blood loss for 1.5 hours and 300 ml,respectively.No injury to the vertebral artery or spinal cord was observed.All the patients were followed up for 12-24 months,which showed that JOA scores one year after operation was increased to 10-17(average 14.56),with the improvement rate of 71.70%,and that the SAC was average 14.86 mm.The X-ray and SCT scans verified the proper position of the screws,with no internal fixation failure or atlantoaxial redislocation.After 3-6 months,all the patients except for one patient achieved a solid bone fusion.One year after operation,the one patient with no bone graft fusion was removed of the internal fixation system and obtained satisfactory restoration of the rotational function.Conclusions Atlantoaxial pedicle screw system is an effective method for the treatment of the unstable atlantoaxial dislocation post traction,for it has the advantages of stable three-dimension fixation,direct screw placement,intraoperative reduction and high fusion rate.

Objective To investigate the thyroid ultrastructural pathological changes of diabetes mellitus (DM) rats as well as the intervention effects of insulin and aminoguanidine. Methods Totally 87 rats were treated with streptozotocin to establish DM animal models and divided into DM group(n=27),insulin intervention group(n=32) and aminoguanidine intervention group(n=28),25 rats were taken as normal controls. Twelve and 20 weeks after the animal model establishment, animals were sacrificed, thyroid tissue was taken and ultrastructure was observed. Results In the thyroid of DM rats, follicular epithelial cells present as applanate shape, microvilli were depleted, rough endoplasmic reticulum dilated to irregular vesicular. None pinocytotic vacuole and casual primary or secondary lysosome were seen. Follicular cavity was dilated, colloid in the cavity had higher electronic-density. Interstitial edema, capillary base lamian was thickened at different stage. Proteo-substance deposition with granulo-shape, cloud shape or homogeneity appeared. The number of thyroid parafollicular cells increased. But endocrine granule in parafollicular cells was few. When compared with DM group, the thyroid tissue injury of insulin intervention group and aminoguanidine intervention group were lessened to different degree. Conclusion The hypofunctional thyroid follicular cells, large quantity of proteo-substance deposition in the interstitium and increased parafollicular cells of DM rats may be related with hyperglycemia toxicity. Insulin and aminoguanidine treatment have some protection effects.

Objective To evaluate the efficacy of moxifloxacin-based triple therapy combined with rebamipide for Helicobacter pylori (Hp) eradication in type 2 diabetic patients.Methods Two-hundred age and sex matched type 2 diabetic patients with Hp infection and accompanied by gastrointestinal symptoms were assigned into rebamipide group (n =100) and placebo group (n =100).Both groups received moxifloxacin-based triple therapy for 10 d to eradicate Hp.Rebamipide was administrated in rebamipide group and placebo was administrated in placebo group for 30 d.All patients proceeded 14C-urea breath test (14C-UBT) hefore and 7-8 weeks after eradication therapy.The blood sugar indices,gastrointestinal symptom scores and Hp eradication rates were measured and compared.Those who had gained successful Hp eradication in both groups proceeded 14 C-UBT again 12 months after eradication therapy,and the re-infection rates of both groups were compared.Results There were no significant differences in the average fasting glucose,postprandial glucose and HbAlc levels between the rebamipide group and the placebo group before and after therapy (P ＞ 0.05),the gastrointestinal symptoms of the rebamipide group were improved more markedly than those of the placebo group after therapy (Rome Ⅲ Criteria,2.1 ± 0.9 vs.4.4 ± 0.7,P ＜ 0.01).The Hp eradication rate with both intention-to-treat (ITT) and per protocol (PP) analysis of the rebamipide group were higher than that of the placebo group [86.0％ (86/100) vs.73.0％ (73/100),P ＜ 0.05 and 92.5％ (86/93) vs.76.8％ (73/95),P ＜ 0.01].The re-infection rates with both ITT and PP analysis of those who had successful eradication in rebamipide group were lower than that in placebo group [19.8％ (17/86) vs.35.6％ (26/73),P＜0.05and20.5％ (17/83) vs.36.6％ (26/71),P＜0.05].Conclusions Moxifloxacin-based triple therapy combined with rebamipide has a more beneficial effect on Hp related symptoms,a higher Hp eradication rate and a lower Hp re-infection rate for type 2 diabetic patients.

Object To explore the antitumor mechanism of Stellera chamaejasme Linn.(SC).Methods SC containing-serum(SCCS)was derived from mice pretreated with different doses of SC.Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used.Inhibition of proliferation was measured using MTT assay.Morphological assessment of apoptosis was performed with fluorescence microscope.DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry.Expression of bcl-2 protein was measured with immunohistochemistry.Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC(pretreated with SC3,6, and 12 g/kg)for 48h resulted in growth inhibition in a dose-dependent manner.Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced."Apobodies'in the apoptotic cells were observed,'ladder"pattern of agarose gel electrophoresis of DNA from 11.7% to 57.4%.Treatment with SC containing serum decreased the percentage of SGC-7901 cell of bcl-2 protein positive expression from 78.3% to 32.9%.Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.

BACKGROUND: There is no study addressing transplantation of umbilical cord mesenchymal stem cells for the treatment of spinocerebellar ataxia.OBJECTIVE: To study the clinical effect of human umbilical cord stem cells transplantation in the treatment of autosomal dominant spinocerebellar ataxias. METHODS: Spinocerebellar ataxias patients selected from Stem Transplantation Center of the 455 Hospital of Chinese PLA were treated with umbilical cord mesenchymal stem cells transplantation via intrathecal injection. The number of umbilical cord mesenchymal stem cells was 107 per transplantation, once per week, for 4 weeks.RESULTS AND CONCLUSION: Both the total score of the International Cooperative Ataxia Rating Scale and Activities of Daily Living score were significantly decreased at 1 month after transplantation compared with before treatment (P < 0.05). The nerve function was significantly improved and the total effective rate was up to 16.7%. Experimental findings indicate that, transplantation of umbilical cord mesenchymal stem cells via intrathecal injection is a feasible and effective treatment to ameliorate the clinical efficacy of spinocerebellar ataxias patients and improve their quality of life.

Objective To observe the expression of 7 cytokeratins in skin epithelial tumors and to assess their diagnostic value. Methods The expression of 7 cytokeratins, including cytokeratin 7(K72.2), cytokeratin 8(C-51), cytokeratin 10(DE-K10), cytokeratin 14(LL002), cytokeratin 17(E3), cytokeratin 18(DC10)and cytokeratin 19(KS19.1), was assessed by immunohistochemical staining (S-P method) in 54 cases of different skin epithelial tumors and 20 normal skin specimens. Results Of 54 cases studied, 10 were squmous cell carcinoma (SCC), 10 basal cell carcinoma (BCC), 19 hair follicle tumor, 2 sebaceous carcinoma, and 13 sweat gland tumor. The expression patterns and distribution of the 7 cytokeratins were different in different skin epithelial tumors. Most of the tumor cells stained diffusely for cytokeratins in SCC, BCC and hair follicle tumor; different cytokeratins expressed in different parts of each of the subtypes of sweat gland tumors. Conclusions Analysis of a selected group of cytokeratins may be helpful in the diagnosis and differential diagnosis of SCC, basal cell carcinoma and skin adenexal tumor.