To compare the efficiency of methods of DNA extraction from lungs of Pomacea canaliculataused in PCR assay, 80 P.canaliculata collected in field were divided into 8 groups and the lungs of each snail were separated from the soft body. Six methods of DNA extraction from lungs of P. canaliculata were used to extract DNA from lungs, i.e. With Qiagen, Tiangen,and Omega commercial DNA extraction kits, guanidine thiocyanate method, Chelex 100 resin method and Chelex-silica particle method. The 16S rDNA of C.canaliculata was amplified by PCR and the concentration of PCR-products relative to marker was determined in order to evaluate the efficiency of each method. It was demonstrated that each method was valid to extract DNA from lungs used in PCR assay, but the concentrations of PCR-products were different. The concentrations of PCR-products obtained by Qiangen kit, Omega kit, Chelex 100 resin method and Chelex-silica particle method were significantly higher than those of other 4 methods of DNA extraction, in which Qiangen and Omega kits were suitable for small sample size. In term of efficiency and cost, Chelex 100 method and Chelex-silica particle method were feasible for large sample scale, while the guanidine thiocyanate method was preferred due to its fast extraction and low cost, but on account of its toxicity, it is used in urgent status or in large scale of sample extraction.

Ojective To enhance the ability of diagnosis and therapy for occult indirect inguinal hernia. Methods Surgical exploration was performed for patients highly suspected with occult indirect inguinal hernia based on clinical manifestations and physical examinations in the initial stage. At present, herniography was used to confirm the diagnosis. High ligation of the hernial sac, non-mesh methods of open groin hernia repair or tension-free inguinal hernia repair was performed for the patients. Results Forty-four cases were given surgical exploration in the initial stage and 41 cases were confirmed and treated by operation. Misdiagnosis was found in 3 cases and surgical exploration revealed no sign of disease. Relapse occurred in two cases within 5 years. Nineteen out of 30 patients were confirmed by herniography in the last five years and operated on. There was no false-positive herniogram in this series. Follow-up evaluation of the patients showed disappearance of preoperative symptoms. Groin hernia has not recurred so far. Conclusions The ability of diagnosis for occult indirect inguinal hernia can be enhanced by herniography. Blind surgical exploration can be reduced. Herniography is a safe, reliable and practical diagnostic procedure.

To evaluate initial experience with shape memory alloy stent as an alterative to colostomy in patients with intestinal obstruction by colorectal cancer, forty four patients diagnosed with acute and chronic colorectal obstructions from malignant causes were subjected to stent placement. Among them 41 patients achieved clinical decompression. Thirty three cases died within 51～720 days and the other 5 cases were still alive without intestinal obstruction for 1～8 months. Our conclusions are shape memory alloy stent may be useful in the management of terminal or high risk surgical patients for palliative purposes instead of colostomy. Palliative management by stent placement combined with chemotherapy and immunotherapy might prolong the survival time of these patients. Placement of stent for acute colorectal bowel obstructions facilitates preoperative bowel preparation for surgical resection and obviates the need for fecal diversion or on table lavage.

Objective To detect and differentiate six major human herpesviruses DNA by PCR, RFLP, DNA clone and sequence analysis. Methods Based on the sequence of well coonserved regions of the DNA polymerse gene in human herpesvimses, we synthesized two pairs of primers, including one pair designed to amplify herpes simplex virus type 1 and 2, Epstein Barr virus and cytomegalovirus, other pair of primer to varicella zoster virus and human herpesvirus 6 by PCR. Identification of the virus species was achieved through restriction enzyme digestion with BamHI and BstUI. Results The products of six human herpesviruses after PCR amplification were from 510bp to 592bp and allowed characterization of herpesvirus type with restriction endonulease analysis. The sensitivity could reach 0.1fg DNA and had no cross reaction to human genomic DNA, bacteria, fungas and virus. 89 cerebrospinal fluids(CSF) and 75 blood specimens were tested for the presence of hepersvirus DNA by this PCR assay, in which 13(14.6%) CSF and 26(34.7%) blood specimens were positive. Comparatively, 10(13.3%) were positive by ELISA for HSVⅠ/Ⅱ,EBV,CMV IgM in blood specimens and significantly lower than that of the PCR rate (P

Objective To observe any rehabilitative effects of individualized exercise training in the treatment of rotator cuff injury in elite table tennis players.Methods Forty table tennis players from the Chinese National Team were studied.Twenty (the experimental group) had rotator cuff injuries and 20 without the injury formed the control group.An individualized rehabilitation treatment protocol was prepared for each of the players in the experimental group and implemented twice weekly for a total of 8 weeks.There was no intervention for the control group.Before and after the treatment,both groups were assessed using a questionnaire for disabilities of the arm,shoulder and hand (DASH),a motion assessment,and BIODEX isokinetic muscle testing.Results There were significant differences in average DASH scores between the two groups,with the experimental group scoring higher than the controls both before and after the treatment.Before the treatment,at angular velocities of 60°/s,180°/s and 300°/s the peak moments of the ER (external rotation,ER) and IR (internal rotation,IR) muscles and the ER/IR ratio were all lower in the experimental group than among the controls.However,after the treatment the performance of the experimental group had improved in all three tests.Conclusion The ER/IR ratio of the dominant shoulder of table tennis players with rotator cuff injury is lower than that of the players without the injury,but individualized rehabilitation treatment can effectively increase the ratio and help to improve their shoulder function.

Objective To summarize the clinical effects of one stop hybrid approach for treating the tetralogy of fallot complicated with aorta pulmonary collateral arteries. Methods From Janu 2008 to June 2013, 79 cases of tetralogy of fallot were complicated with aorta pulmonary collateral arteries with mean age (5.4±3.9) years and weight (18.2±5.7) kg. All patients were diagnosed by echocardiography examination and accepted 64-row CT for demonstration of no dysplasia in pulmonary ressels, McGoon index was (1.7±0.6) and Nakata index was (176.7±7.3). 76 cases had aorta pulmonary collateral arteries for preliminary screening, and accepted cardiovascular angiography before operation. 3 cases were misdiagnosed before surgery and were confirmed by cardiovascular angiography after operation. Results There were 4 deceased cases with an operative mortality of 5.1%. 1 case died of lung infection and 3 cases died of refractory heart failure. In all aorta pulmonary collateral arteries, the smallest diameter was 2.5 mm, the largest was 9.4 mm, average (5.3±2.1) mm. 2 to 21 coils were implanted in respective case with an average of 10.2 coils per case. The maximum number of coils implanted in 1 single ressel was 9. The average number of ressels occluded was 3.7 (range from 1 to 11 ressels) in each case. One case received second closure due to re-open of collateral arteries on neo-vasculanigation. 14 cases had lung infection and 3 cases had pulmonary edema post operation. 75 cases were followed up for 3-72 months. 4 cases with post operative heart function of NYHA gradeⅢtoⅣand all other cases with gradeⅡon higher. Conclusions Peri-operative hybrid management for aorta-pulmonary collateral arteries can increase the operative success rates and reduce complications.

BackgroundHigh myopia is one of leading causes of blindness,so far the pathogenesis remains unclear.Two single-nucleotide polymorphisms (SNPs) of rs6885224 and rs12716080 in CTNND2 gene were recently found to be associated with high myopia in Singaporean Chinese.But whether these SNPs are related with the pathogenesis of high myopia in Han Chinese is worth studying,Objective This study was to investigate the relationship between the genetic variations of the CTNND2 gene and high myopia in Han Chinese.MethodsA case-controlled association study was designed.Nine hundred and thirty-three individuals with high myopia and 1227age- and gender-matched normal subjects were included in this study.The 5 ml of periphery blood was obtained from all subjects for the extraction of genomic DNA.The target DNA was amplified using PCR and purified by the SNaPshot method.Four SNPs rs12716080,rs917012,rs6885224 and rs16901340 in the CTNND2 gene were genotyped.This study was approved by the Ethic Committee of Sichuan Provincial People Hospital.Written informed consent was obtained from each subject before his/her enrollment.Results The frequencies of the genotypes rs6885224,rs12716080,rs917012,rs16901340 SNPs were in Hardy-Weinberg equilibrium (HWE) ( P=0.181,0.085,0.732,0.313,0.264,0.663,0.084,0.196).There were no significant differences in genotypes frequency distribution ( in turn P =0.654,0.406,0.828,0.403 ) and allele frequency distribution of the CTNND2 gene ( in turn P =0.377,0.209,0.743,0.198) between the high myopia group and normal control group.The haplotypes (TA and GA)frequencies of rs12716080 and rs917012 in the high myopia group were significantly different from those of the normal control group(TA:0.784 vs.0.719;GA:0.087 vs.0.136) (x2 =6.115,P=0.013 ;x2 =6.634,P=0.010),but those of GG were similar between the high myopia group and normal control group ( 0.123 vs.0.143,x2 =0.889,P =0.346). ConclusionsThe SNPs rs12716080,rs917012,rs6885224 and rs16901340 in CTNND2 gene were not responsible for high myopia,however,the haplotypes of rs12716080 and rs917012 are susceptible for high myopia in Han Chinese.

BACKGROUND: It is proved that nerve regeneration induced by terminolateral neurorrhaphy(TLN) is not as active as that induced by end-to-end suture. Exogenous epidermal growth factor(EGF) increases the opportunity of neuron survival in vitro and promotes nerve regeneration. Whether it can increase nerve regeneration after terminolateral neurorrhaphy deserves further study.OBJECTIVE: To evaluate the effect of exogenous EGF in promoting nerve regeneration after terminolateral neurorrhaphy.DESIGN: A randomized controlled trial.SETTING: Orthopedic Institute of Chinese PLA General Hospital.PARTICIPANTS: The trial was conducted in the Orthopedic Institute of Chinese PLA General Hospital from September 2001 to February 2002. A total of 32 male Wistar rats, weighting 200- 250 g, were randomized to control group and EGF group with 16 rats in each group.METHODS: The right peroneal nerve was transected and an epineural window of 1 mm was created on the neighboring tibial nerve. The distal end of the transected peroneal nerve was sutured to the windowed tibial nerve by means of end-to-side attachment. Each rat in EGF group received injection of 0. 1 mL/d EGF diluted with normal saline at 2 g/L for two weeks while each in control group received injection of normal saline (0. 1 mL/d) at the distal site of the transected peroneal nerve for two weeks. Histological, morphological and electrophysiological examinations were performed 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: The regeneration rate of myelinated nerve, motor nerve conduction velocity and ultrastructural changes of the two groups.rate of myelinated nerve fibers: 4 and 8 weeks after operation, it was better in EGF group[ (52.42 ± 1.45)% and(61.41 ± 1.54)% ] than that in control nerve conduction velocity: 4 and 8 weeks after operation it was obviously greater in EGF group[ (30. 33 ±0. 88)m/s and(34. 36 ± 1.09)m/s] than that in conObservation of ultrastructure: The number of myelinated nerve fibers, and the thickness and maturation degree of myelin sheath were significantly better than those in control group.CONCLUSION: Exogenous EGF can promote nerve regeneration, increase nerve conduction velocity after terminolateral neurorrhaphy.

Objective To evaluate the primary result of proximal femoral nail anti-rotation(PF-NA)in treating osteoporotic intertrochanterie fractures. Methods Between March 2006 and August 2007.PFNA or Gamma nail intemal fixation were performed in 31 patients with osteoporotic intertrochan-tefie fractures.of whom were 19 males and 12 females at a mean age of 75.6 years(45-89 years).Ac-cording to AO classification,there were 8 type A1 fractures,16 type A2 fractures and 7 type A3 fractures.Aeearding to Evans-Jensen classification.18 cases were rated as Evans Ⅱ and 13 as Evans II.AIl frac-tares were closed ones and 6 patients were combined with internal diseases.PFNA was implanted throush a mini-incision without exposure of the fracture section during operation in 12 patients. Results Of all.6 patients received emergency operation,19 did operation within 3 days and 6 waited for selective op-eration(5-7 days)after controlling internal diseases.PFNA had shorter operation time,less hlool loss and shorter union time compared with Gamma nail(P＜0.05).No complications such as infection,non-union or loosening of nails were found.According to HarriS hip rate scale system,the excellence rate of PFNA was 83.4%and that of Gamma nail 73.7%. Conclusion PFNA is a good choice to treat osteo-perotie intertrochanteric fractures.owing to its advantages of stable fixation,minor trauma,eaSy opera-tion.short operation time,little bleeding,minor lOSS of selerotin and early exercise.