Objective To investigate the relationship between different expression of gastrocolic omentum visfatin (VF) mRNA and gestational diabetes mellitus (GDM) in late trimester pregnancy. Methods One hundred cases of pregnant women in late trimester were divided into two groups, including 45 GDM women( GDM group) and 55 normal glucose tolerance women( NGT group). Semi-quantitative RT-PCR was applied to determine the expression of VF mRNA in gastrocolie omentum. Fast-blood glucose, insulin,cholesterol(TC) and triglyceride (TG) levels were measured. The progestational body mass index(BMI) and the homeostasis model assessment(HOMA) insulin resistance(IR) were calculated in these cases. Results The expression of gastrocolic omentum VF mRNA in GDM group and NGT group were 0.8 ±0.4,0.5±0.3. Fast-blood glucose levels were(4.12±0.14),(3.65±0.13) mmol/L. Insulin levels were (72±5) ,(61±5) pmo]/L TG levels were (5.6±0.3), (3.8±0.3) mmol/L TC levels were(5.6±0.9 ), (3.9±0.3) mmol/L HOMA-IR were 12.5±5.9,9.5±0.8. The progestatianal BMIs were(22.6±0.8), (20.9±0.4) kg/m2. All the indices in the GDM group were significantly higher than NGT group (P<0.05). Furthermore, the expression of gastrocolic omentum VF mRNA had a positive correlation with BMI (r=0.32, P<0.01), but had no association with insulin resistance or metabolic parameters. Conclusion The experiment demonstrates that different expression of VF mRNA has a strong correlation with GDM and obesity.

Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.

Objective To explore operation method,skills,and curative effect of orthophofia microscope discectomy(OMD)for the treatment of lumber disc herniation.Method Retrospective analysis of the 64 patients(74 herniated discs)of lumbar disc herniation heated by OMD,which included 38 segments posterior-lateral extrusion,32 segments central type herniated,and 4 segments extra-lateral extrusion.Resuits All patients were followed-up 6 to 8 months,average 7 months,54 cases were good and 4 cases were fair and no case was bad according to the Nakai scale.The choiceness rate was 93.75%,and no serious complications.Conclusions Orthophoria microscope discectomy has a better third dimension,good operation field of vision,less invasion and best clinical curative effect.Orthophoria microscope discectomy may be the best way of discectomy for the treatment of lumber disc hemiation.

Objective To detect gene mutations associated with dapsone-,rifampicin-and ofloxacinresistance in lesions of patients with recurring or treatment-resistant leprosy collected from 2010 to 2011.Methods Clinical data and lesional specimens were collected during 2010-2011 from patients with recurring or treatment-resistant leprosy who were diagnosed and reported by provincial centers for leprosy control.Mycobacterium leprae DNA was extracted from the specimens and subjected to PCR for the amplification of folP1,rpoB and gyrA genes.The PCR products were directly sequenced and BLAST program was used to compare the sequence of isolated strains with the reference sequence in GenBank.Results Twenty-four patients were enrolled in this study,including 13 with recurring leprosy and 11 with treatment-resistant leprosy.Twenty-one patients showed positive PCR results in all the three regions.Of these PCR-positive specimens,3 from 1 patient with recurring and 2 patients with resistant leprosy harbored a point mutation,acc (threonine)→gcc (alanine),at codon 53 in the floP1 gene,1 from a patient with recurring leprosy harbored a missense mutation,gat (aspartic acid) → aac (asparagine),at codon 441 in the rpoB gene.Conclusions Mutations are detected in the folP1 and rpoB genes,which are associated with the resistance to dapsone and rifampicin respectively,but not in the ofloxacin resistance-associated gyrA gene,in Mycobacterium leprae isolates from patients with recurring or treatment-resistant leprosy.

Objective To explore the applied value of multi-slice spiral CT(MSCT) scan in the traumatic facial palsy.Methods 41 cases of traumatic facial palsy were scanned using MSCT,the MPR of interseted region of temporal bone and curved planar reconstruction(CPR) images were performed on Philips Mx8000 MSCT work station.The types,locations and course of the temporal bone fracture and the involving features of the external auditory canal,middle ear,inner ear were observed.Results 39 cases of the 41 patients had temporal fracture in different location.34 cases were longitudinal fracture,2 were transverse fracture and 2 were mixed fracture.The facial nerve canals were involved in 21 cases.Conclusion MSCT and the reformation images can help to study the facial nerve canal in the traumatic patients.It is important to use the MPR and CPR in the traumatic facial palsy.

BACKGROUND: Treatment of large bone defects is an important problem faced by orthopedic physicians. Allogeneic bone transplantation is a classic method, but it has many restrictions. The membrane guided bone regeneration technique has become an important method for the research nowadays.OBJECTIVE:To compare the effects of self-made high strength biodegradable nano-hydroxyapatite/multi(amino acid) copolymer (n-HA/MACP) guided bone regeneration membrane tube and allograft bone graft segment in the repair of large segmental bone defect in goats. METHODS: The model of 30 mm large segment bone defect in the middle section of the femur in 32 adult goats was established. Experimental group used self-made n-HA/MACP tube to bridge defects following bone plate fixation. The control group was treated with allograft bone graft bone segments combined with plate fixation. The animals were sacrificed at 4, 8, 12 and 16 weeks after operation, and the bone callus growth was observed in the specimens. The X-ray and histological observations were performed at the same time. Biomechanical measurement of plate fixation of allograft cortical bone segment was done at 12 and 16 weeks after operation. RESULTS AND CONCLUSION: (1) After 4 to 16 weeks, gross and pathological results showed fibrous callus growth inside and outside of the membrane tube, and the fibrous callus gradually hardened into a bony callus. Additionally, the callus was larger in the experimental group than in the control group. X-ray films showed significantly increased lateral callus density in the experimental group as compared with the control group. (2) Maximum bending strength was significantly higher in the experimental group than in the control group at 16 weeks (P < 0.05). Overall, the n-HA/MACP membrane tube used for bridging large segment bone defects in goats can obtain similar repair effects to the allograft cortical bone, and further achieve the better mechanical strength of the new bone segment than the allograft bone.

Objective To investigate the prognostic value of CDKN2A mRNA in glioblastoma (GBM).Methods CDKN2A gene mRNA data were obtained from three different GBM database online (TCGA,REMBARNDT and GSE16011).The correlations between overall survival (OS) and CDKN2A expression were analyzed by Kaplan-Meier method and multivariate Cox regression analysis.Results In the TCGA database (n =358),patients with high CDKN2A mRNA level got longer OS than those with low expression level [median OS:18.0 months (95 ％ CI 15.0-21.0 months) vs 13.9 months (95 ％ CI 12.4-15.4 months),P =0.001].In another two validation datasets,patients with high CDKN2A mRNA level had longer OS than those with low expression level [median OS in REMBRANT:16.6 months (95 ％ CI 13.3-19.8 months) vs 11.8 months (95 ％ CI 7.3-16.4 months),P =0.019; in GSE16011:11.9 months (95 ％ CI 8.3-15.6 months) vs 8.4 months (95 ％ CI 6.2-10.5 months),P =0.005].CDKN2A mRNA level was an independent prognostic factor for GBM.The combination of CDKN2A mRNA expression with MGMT promoter methylation status or G-CIMP status/IDH1 mutations provided an optimized prognostic factor in GBM patients.Conclusion The CDKN2A mRNA has prognostic value in GBM patients,which provided an optimized stratification strategy based on multiple biomarkers.

BACKGROUND: Pathophysiological mechanism of local microenvironment is complex after central nerve injury; especially,both inflammatory reaction at an acute phase and formation of secondary glial scar have tremendous effects on effective regeneration of axon, regeneration and arrangement of local nerve cells, proliferation and migration of local stem cells;therefore, it becomes a basic reason for blocking nerve repair in an early period. Thus, how to effectively resist inflammatory factors in injured region at an acute phase and how to optimize microenvironment of neural regeneration are the most important strategies for repairing spinal cord injury in recent years.OBJECTIVE: To design, establish and screen the best expression of interleukin-6 receptor (IL-6R) α to inhibit shRNA adenovirus expression vector by using spinal cord injury models.DESIGN: Duplicative measurement study.SETTING: Department of Spine Surgery, the Second People's Hospital of Shenzhen.MATERIALS: A total of 40 healthy Wistar rats, either gender, 8-10 weeks old, were selected in this study. Rabbit-anti-rat glial fibrillary acidic protein (GFAP) antibody Ⅰ was provided by Santa Cruz Compan; siRNA eukaryon expression plasmid pGenesil (cohtaining green fluorescent expression system) was provided by Wuhan Jingsai Bioengineering Company.METHODS: The experiment was carried out in ImmuneOpening Laboratory, Basic Medical Faulty, Tongji Medical College, Huazhong University of Science and Technology, and Medica Laboratory Center, the Second People's Hospital of Shenzhen in November 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of IL-6 receptor (IL-6R) α target sequence with 9 bp spacer were designed and synthesized, then the recombinant adenovirus expression vectors with green fluorescence protein were constructed in vitro respectively. The acute spinal cord injury models were completed, and the adenovirus recombinants were regionally injected post 12 hours after spinal cord injury;in addition, the inhibitory effect of RNA interference (RNAi) on expression of IL-6R in local region after spinal cord injury were detected by using real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot so as to screen adenovirus expression vector which had the best inhibitory effect on expression of IL-6R.MAIN OUTCOME MEASURES: Inhibitory effect of RNAi on expressions of IL-6R RNA and protein in local region after spinal cord injury.RESULTS: Sequence analysis showed that IL-6R-shRNA recombinant adenovirus expression vector was successfully constructed, and optimal IL-6R-shRNA recombinant adenovirus expression vector was screened by using real-time fluorescence quantitative RT-PCR and Western blot. The IL-6R expressions were 49% and 56% at the levels of mRNA and protein, respectively.CONCLUSION: The IL-6R--shRNA recombinant adenovirus expression vector is successfully constructed and screened.The gene expression of IL-6R can be highly inhibited after acute spinal cord injury.

objective To explore the indication of mechanical ventilation and evaluate the effects of mechanical ventilation with the least delay on the Severe cases of hand-foot-mouth disease(HFMD).Methods Retrospective cohort study was conducted among the severe HFMD cases(n=66)admitted to pediatric intensive care unit(PICU)between July 2008 and september 2009.Sixty-six cases were divided into two groups:42 cases(group A,July 5 to October 31,2008)were ventilated with the common mechanical ventilation,24 cases(group B,April 21 to september 30,2009)were ventilated at least delay when the symptoms of neurogenic pulmonary edema appeared.Then the diffcrences on the survival rate and the mortality in the group A and group B were investigated.Results Twenty out of 42 patients died(47.6%),eighteen were cured(42.6%) and four showed improved signs(9.5%)in group A.Twenty-two out of 24 patients were cured(91.7%),two showed improved signs(8.3%)and no death in group B.The clinical effect of group B was much better tllan group A(P<0.001).Conclusion Early mechanical ventilation would improve the survival rate and decrease the mortality of severe hand-foot-mouth disease.

BACKGROUND:Bisphosphonates that can suppress osteoclast activity strongly is a powerful inhibitor for bone resorption, which has been reported to have good effects in the treatment of fibrous dysplasia.
OBJECTIVE:To evaluate the clinical effect of surgical treatment combined with bisphosphonate therapy in the treatment of fibrous dysplasia.
METHODS: Fifteen patients with fibrous dysplasia of the long bone were subjected to surgical treatment and oral bisphosphonate therapy postoperatively. Limb pain, limb function, local X-ray manifestations and blood alkaline phosphatase activity were observed before and at 3, 6, 12, 24 months after treatment. The therapeutic effects were evaluated using modified Macnab standards.
RESULTS AND CONCLUSION:Al the 15 patients were folowed up for more than 24 months, and their pain symptoms were significantly relieved after the combined treatment of surgery and bisphosphonates, excelent in 12 cases and effective in 1 case, and the total efficiency was 100%. X-ray films showed that the density slightly decreased in operation areas after 3 months, but at 6 months after treatment, the cortical bone was thickened and the marrow cavity density increased in the operation areas, and non-focal zone was continuously enlarged. No fractures and recurrence were found in al patients. At 6 months after treatment, the serum alkaline phosphatase activity decreased significantly (P < 0.05). The results suggest that surgical operation with bisphosphonate treatment for bone fibrous dysplasia can effectively inhibit the bone resorption, promote bone formation, increase the intensity of the lesion, reduce fractures and recurrence rate, so as to achieve a better therapeutic effect in clinic.