Objective: To establish a stable, reproducible, and suitable reaction system of Anoectochilus formosanus for ISSR analysis of genetic differences according to the ISSR-PCR characters in plants of Anoectochilus Bl. Methods: The initial ISSR reaction system in plants of Anoectochilus Bl. was established and then the primers were screened. After that, PCR amplifications were carried out until the different concentrations of the factors in the ISSR reaction system were designed, and based on the principle of high stability and reproducibility, the stable and reliable ISSR reaction system was eventually established after a series of adjustment and optimization of the reaction system. Results: The optimal ISSR reaction system in plants of Anoectochilus Bl. was reported for the first time, and the volume of 25 μL contained approximately 20 ng template DNA, 1 U Taq DNA polymerase, 0.2 mmol/L deoxyribonucleotide triphosphate (dNTP), 2 mmol/L MgCl2, 200 umol/L dNTP Mixture, and 0.2 μmol/L primer within 1x reaction buffer [10 mmol/L Tris-HCl (pH 8.3), 50 mmol/L KCl]. The reaction mixtures were denatured at 94°C for 7 min and subjected to 45 cycles of 30 s at 94 °C, 45 s at 52 °C, 2 min at 72 °C, and a final extension step of 10 min at 72 °C and eventually stored at 4 °C. Conclusion: The ISSR-PCR systems established for studying on the plants of Anoectochilus Bl. show the characters of stable reaction system, better repeatability, clear marker site, and reliable abundant polymorphisms. It is suitable for the study on genetic diversity in plants of Anoectochilus Bl. in difference wild populations and genetic variation of its sterile seedlings after long time tissue culture.

Objective To investigate the protective effects of mild hypothermia on cells in the CA_1 region of the hippocampus in gerbils following global ischemia-reperfusion injury(IRI),and to explore their mechanism. Methods IRI models were established in 75 gerbils.Any changes in TUNEL positive cells and the expression of Bax and Cytochrome C were then observed in normothermic(N) ,hypothermic(H)and sham(S) groups through immu- nohistochemistry methods.Results In the H group(as compared with the N group)apoptotic cells in the CA_1 sub- field of the hippocampus significantly decreased.The expression of Bax and Cyt C at 3 h,6 h and 1 d were de- creased,and the expression fastigium was delayed.Conclusion Mild hypothermia can moderate and delay cell ap- optosis,and its mechanism might be related with reducing and delaying the expression of Bax and Cyt C released by mitochondria.

Genomics ; Medicine, Chinese Traditional

Objective To explore the high risk factors, pathogenesis and methods of early diagnosis of periventricular leukomalacia(PVL) in premature infants.Methods The history of intrauterine hypoxia-ischemia was investigated in premature infants;TORCH-IgM antibodies in cord blood of premature infants were measured by ELISA; Nitric oxide (NO) levels in their cord blood were determined by nitric acid reducing enzyme means. Results Thirty-nine of 52 premature infants in PVL group had a history of intrauterine hypoxia-ischemia; TORCH-IgM antibody positive rate in cord blood of premature infants in PVL group was significantly higher than that in control group(P

Objective To evaluate the outcome of dorsal double locking compression plate (LCP) in treatment of stretched unstable distal radial fractures.Methods Fourteen cases of stretched unstable distal radial fractures were treated by reduction and internal fixation using straight or L-shaped anatomic LCP via dorsal approach.Wrist functional exercise was conducted immediately after operation.Follow up was made after operation to assess motion pain,functional score of wrist,and complication incidence at postoperative 12 weeks,24 weeks,and 1 year.Results Follow-up was lasted for 5-24 months.At postoperative 12 weeks,24 weeks and 1 year,mean visual analogue scale (VAS) was (1.88 ±0.26) points,(0.87 ± 0.14) points and (0.37 ± 0.06) points respectively and wrist functional score (Gartland-Werley score) was (6.45 ± 1.72) points,(2.73 ± 0.52) points and (2.10 ± 0.31) points respectively.According to Garfland-Werley score in the latest follow-up,the results were excellent in 10 cases,good in two,and fair in two.Besides,one case was combined with myotenositis of extensor pollicis longus muscle tendon and another case with myotenositis of extensor tendon.Conclusion Dorsal double LCP is one of the effective methods for stretched unstable distal radial fractures that can reconstruct anatomical structure of the wrist efficiently and attain satisfactory functional recovery,with no obvious pain.

OBJECTIVETo evaluate the clinical results of the medial patellofemoral ligament (MPFL) reconstruction combined with the lateral retinacular release for the treatment of recurrent patellar dislocation.

METHODSFrom March 2011 to June 2013, 15 patients with recurrent patellar dislocation underwent arthroscopic MPFL reconstruction combined with the lateral retinacular release. The graft was autogenous semitendinosus and semimembranosus tendon. There were 5 males and 10 females with an average age of 19.4 years old (ranged,14 to 32 years old). The patients suffered recurrent patellar dislocation at least twice preoperatively. Preoperative conventional X-ray, CT, and MR examination were used to analyze the causes of the patellofemoral joint and MPFL injury. Preoperative Lysholm score was 69.85 ± 11.52. During operation, the arthroscopic examination was performed to evaluate the patellofemoral alignment and patellar tracking.

RESULTSAll the patients were followed up for an average of 27.6 months (ranged,12 to 36 months) with no recurrent dislocation and sub-dislocation. All the patients showed negative apprehension test at straight and 30 ° flexions of knee. The range of motion of knee returned to normal level at 12 months after operation. There were no patients with subjective discomfort of knee. Postoperative Lysholm score was improved to 92.60 ± 5.75.

CONCLUSIONThe technique of arthroscopic MPFL reconstruction combined with the lateral retinacular release is an effective surgical procedure for the treatment of recurrent patellar dislocation, which can relieve the symptom of knee and improve the patella stability and knee function.

Adolescent ; Adult ; Arthroscopy ; Female ; Humans ; Knee Joint ; surgery ; Male ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; physiopathology ; surgery ; Patellofemoral Joint ; physiopathology ; surgery ; Range of Motion, Articular ; Treatment Outcome ; Young Adult

Objective To evaluate arthroscopic PCL (posterior cruciate ligament) reconstruction with an implant fixaliun of hamstring tendon knot and bottle-neck technique.Methods Thirteen cases with PCL injury were treated with arthroscopic PCL reconstruction with an implant fixation of hamstring tendon knot and bottle-neck technique between April,2002 to June,2005.The patients were followed up fur at least seven months to evaluate the clinical effects by means of ROM of the joint,post drawer test,Lachman test,Lysholm and IKDC (International Knee Documents Committee) scores.Results The patients were followed up for an average of 13.4 months.No limitation of knee extension was found six months after operation.Eleven patients had normal knee flexion(＞140?), one had knee flexion of 120?and one 100?.In posterior drawer test,11 patients were gradeⅠand two gradeⅡ.In Lachman test,11 patients were rated as within gradeⅠand two gradeⅡ.By Lysholm knee function scale,the average knee scores were 90.47?4.13 and 78.82?2.46 respectively before and after operation,and the dif- ference was of statistical significance (t=2.416,P=0.029).By IKDC scale,six were grade A,five grade B and one grade C.Conclusion Arthroseopic PCL reconstruction with an implant fixation of hamstring tendon knot and bottle-neck technique is a reliable method to restore knee function and will have a prospect of wide application be- cause of its good tissue biocompatibility,less invasion,and less operative expenditure.

Aim The kinetics of nicotinic acetylcholine receptor (N-AChR) was stadied. MethodsThe saturation experiments of Torpedo electric organ N-AChR were taken with125I-?-BuTX. The results were calculated by Scatfit program. Experiments were thenperformed to study the kinetics of dissociation of ?-BuTX and nicotine from N-AChR,respectively. When their binding reached saturation, an excess of 1000 times ?-BuTX ornicotine was added. The saturation binding properties of the N-AChR extracted fromleg muscles of the 13th day chick embroys and optic lobes of the 20th day chick em-broys were studied. The results were calculated by Scatfit program. The othe experi-ments were taken to observe the competition between mcotine and 125I-?-BuTX forbinding with Torpedo, optic lobe and skeletal muscle of the chick N-AChRs. ResultsThe saturation experiments of Torpedo electric organ N-AChR resulted in a Scatchardplot of hyperbola which responded to the model of two kinds of receptor with on eli-gand.The difference of and B max between high affinity binding site and low affnitybineing site was significant. The dissaciation experiment showed that the fast dissocia-tion rate of tow ligands was 500 times more than that of the slow dissociation rate. Thisresult suggested that there may be two subtype N-AChRs.The saturation bindingproperties of N-AChR of leg muscles and optic lobe of the chick embroys revealed aScatchard plot of two kinds of N-AChR indicating a single type of site. The bindingaffinity of receptor of optic lobe was 100 times more than that of muscles. In competi-tion for Torpedo receptor by nicotine and 125I-?-BuTX, the values of IC50 were different:which suggested that two kinds of receptor sites were existent. In competition ofNicotine and 125I-?-BuTX for optic lobe and skeletal muscles of the chick N-AChR, thevalue IC50 of skeletal muscles N-AChR was 7. 7 times higher than that of optic lobe. Itindicated that two kinds of N-AChR subtype existed in the optic lobe and in theskeletal muscles of chick respectively. Conclusion N-AChR of Torpedo electric organcontains two kinds of subtype receptors. N-AChR of optic lobe of chick embroys is onesubtype receptor, and N-AChR of skeletal muscles of chick embroy is another subtypereceptor.

Objective To investigate the three-dimensional structure of normal male canine prostat- ic duct and acinus system,and to study the mechanism of intraprostatic urinary reflux(IPUR)resulting from high pressure of the posterior urethra by experiment.Methods Using scanning electron microscopy (SEM)and corrosion casting methods,high pressure of the prostatic urethra was produced based upon IPUR in normal male canines.Acrylonitrile-butadiene-styrene(ABS)casting solution was injected into 8 Beagle canines'normal prostates to induce intraprostatie resin solution reflux.Results Six casting molds of nor- real canine prostate specimens were obtained,and the casting failed in 2.By corrosion and casting treatment, the prostate appeared to consist of several glandular lobes.The columnar apophyses of glandular surface were observed.The most glandular ducts pointed outward from urethral peristome.Each glandular duct system was made up of many bifurcated branches from the bough as a duct tree.More than 90% of the bifurcations were Y-shaped,and 6 bifurcated branches were rarely seen.The tip of the duct was vesicular.The diameter and length of the bough were(0.37?0.14)mm and(1.14?0.04)mm,respectively.The intersectional angle of urethra and peripheral glandular duct was the biggest.Peripheral glandular duct was perpendicular or con- verse to urinary flow.Peripheral glandular aeinus appeared elliptic and velvet.The mesh-like and crater-like depressions were shown on the surface of central glandular aeinus.Ejaculatory ducts independently and branchlessly entered the urethra.Conclusions The resin corrosion casting methods combined with SEM can clearly show the three-dimensional structure of normal male canine prostatic duct and acinus system. IPUR may easily occur in the peripheral glands from the morphological view.It is suggested that prostatitis occurs more commonly in peripheral glands,confirming a fact that high pressure of the posterior urethra can induce IPUR.

ObjectiveTo explore the mechanism of estrogen inhibiting osteoblast apoptosis induced with serum hungry.MethodsOsteoblasts of the second or third generation from newly born SD rats calvaria were divided randomly into three groups: control group,serum hungry group,serum hungry with estrogen group.Cells of each group were incubated for 1,2,3,5,7 or 14 d,and then were stained immunohistochemically.The rates of positive cells of each group were analyzed.ResultsThere was a little positive expression of Bax,Bcl2 and Fas in control group.The expression of Bax and Fas were significantly increased(P<0.05)in serum hungry group,peak time was 14 d,but the expression of Bcl-2 were not affected.Compared with that of serum hungry group,the expression of Bax and Fas significantly decreased(P<0.05) in serum hungry and estrogen group,peak time was still 14 d,while that of Bcl-2 increased(P<0.05).ConclusionSerum hungry can increase the expression of Bax and Fas in osteoblast,that can be inhibited by estrogen.Estrogen can also increase the expression of Bcl-2 in osteoblast.All of these may play a role in inhibiting osteoblast apoptosis induced with serum hungry.