Intracytoplasmic sperm injection,which combined with in vitro fertilization and micromanipulation techniques,has been applied to research the molecular mechanisms of fertilization,and produce the sexing livestock and transgenic animals.The research advances in porcine intracytoplasmic sperm injection,including in vitro maturation and pretreatment of oocytes,selection and treatment of spermatozoon,artificial activation of injected oocytes after injection,and improvement of the operation technique were reviewed.

Objective To analyze postoperative effect of a new rectopexy technique with Douglas pouch elevation for complete rectal prolapse (CRP) in adults.Methods From January 2010 to May 2011, 52 CRP cases were treated by rectopexy with Douglas pouch elevation.In terms of different surgical techniques, patients were divided into two groups : Group A (n =28) received laparoscopic rectopexy with Douglas pouch elevation;and Group B (n =24) received laparoscopic rectopexy with Douglas pouch elevation combined with procedure for prolapse and hemorrhoids (PPH).Rectal prolapse, constipation, and fecal incontinence in the two groups were assessed respectively before surgery, and on the 6th, 12th, and 24th month after surgery, postoperative complications were evaluated with severe grading of surgical complications.Results Symptoms of rectal prolapse disappeared at half a year after surgery in all patients, and relapsed to different extent afterwards.However, two years after operation, the status of rectal prolapse in Group B tended to be stable.The trend of constipation after surgery was consistent with that of rectal prolapse.In addition, fecal continence improved gradually on half a year after surgery in the two groups and recovered to the optimal status in the first postoperative year.However, in the second year, the results of fecal incontinence reduced slightly in Groups A, and Group B became stable.Two years after surgery, four cases in Groups A relapsed while there was no recurrence in Group B.Difference of Grade I to Grade Ⅲ complications among the two groups was statistically insignificant (x2 =0.05, P ＞ 0.05).Conclusions The clinical effect of laparoscopic rectopexy with Douglas pouch elevation associated with the procedure for prolapse and hemorrhoids (PPH) is better than that without PPH for female and male CRP patients with severe symptoms.

This study was aimed to observe the influence of Catalpol on real-time gait analysis of early recovery after permanent middle cerebral artery occlusion (pMCAO) in rats to evaluate its effect on the improvement after cerebral ischemia. Rats were randomly divided into 6 groups, which were the normal control group, model group, Tianbaoning group, Catalpol 15, 30, 60 mg·kg-1 group. Rats were trained on the gait instrument for 7 days before pMCAO, 3 times/day. After the training, pMCAO model was made. And continuous infusion was performed from the 3rd to the 14th day after the operation. Then, the real-time gait behavior was detected on the 14th day. The re-sults showed that 14 days after the surgery, compared with the normal control group, the models had a significant extending in the duty cycle (P < 0.01), and obvious increasing of the average body angle of absolute value (P <0.05), the shortening of the two feet supporting time (P < 0.05), and extending of three feet supporting time (P <0.01), and increasing of the coordination index (P < 0.01). Compared with the model group, the Catalpol of 30 mg·kg-1 and 60 mg·kg-1 group has obviously decreased duty cycle and average body angle of absolute value (P <0.05, or P < 0.01). The Catalpol of 30 mg·kg-1 can obviously reduce the coordination index of the right front foot relative to the other three feet (P < 0.05), which improved the coordination of cerebral ischemia animals. It was concluded that Catalpol can improve the real-time gait behavior changes of cerebral ischemia model rats. There-fore, Catalpol have a neural protective effect on cerebral ischemia.

Objective To investigate the effect of Curcumin on rats with severe acute pancreatitis associated renal injury and explore the possible mechanisms.Methods A total of 72 rats were randomLy divided into sham-operated group(S group, n =24), severe acute panceratitis with renal injury group(M group, n =24) ,Curcumin-treated group(Cur group,n=24).The S and M groups were given 1.5 ml saline through intragastric administration 3 hours before operation,while the Cur group was fed with same amount of Curcumin diluent(200 mg/kg).The pancreas and pancreatic tail-segment was dissociated and the head of pancreas was occlused in rats to form the model,blood vessel forceps was loosed after 3 hours.All the rats were sacrificed at 12 h,24 h, 36 h and 48 h after modeling.The level of ascites, serum amylase, creatinine, blood urea nitrogen were detected and the pathological chang of pancreas was observed under light microscope.Take the right kidney for Superoxide Dismutase (SOD) determination and the expression of hypoxia inducible factor-1αmRNA in the right kidney was detected with realtime polymerase chain reaction (RT-PCR).Results Compared with Cur group, the level of ascites, serum amylase, creatinine, blood urea nitrogen in M group were significantly increased, but the activity of SOD, the express of hypoxia inducible factor-1αmRNA had a significant decline (P＜0.05).The tissue damage of pancreas and the kidneys became more serious.But a better performance was to be found in the function of the Cur group's kidney and pancreas, and the the difference was statistical significance (P ＜0.05).Conclusion Curcumin has a good protective effect on severe acute pancreatitis associated renal injury.It may be through up-regulation expression of HIF-1α mRNA and increase the activity of superoxidase dismutase, then reduce the cell apoptosis and necrosis of the kidney, improve the ability of the kidney to tolerate hypoxia.

Objective To provide the experimental basis for the further research of the interacting proteins with Stathmin ,the Stathmin gene Pichia pastoris expression system was constructed ,the expressed Stathmin product was purified and identified .Meth‐ods Stathmin gene was amplified from tumor cell line of SKBR3 by PCR method and cloned into the yeast expression vector pPIC3 .5K .The recombinant vector pPIC3 .5K‐Stathmin was constructed and transformed into Pichia pastoris GS115 .The positive clones were screened by YPD medium containing Geneticin 600 μg/mL .Expression was induced with 0 .5% methanol and expres‐sion products were identified by SDS‐PAGE and Western Blotting .Results DNA sequencing result showed that the gene fragment was consistent with Stathmin gene sequence .pPIC3 .5K‐Stathmin was selected from YPD culture medium containing Geneticin ,and the positive clones were identified by PCR .SDS‐PAGE showed that a 37 × 103 protein band could be seen on the PAGE gel after Coomassie Blue staining ,which was further confirmed and identified as Stathmin protein by Western Blotting .Conclusion Stathmin yeast expression vector is successfully constructed and expressed in Pichia pastoris ,which laid the foundation for the study of inter‐acting proteins with Stathmin ,and for the preparation of the biological treatment drugs of Stahtmin target .

Objective To observe the efficacy and toxicity of low dose thalidomide combined with VAD regimen contained one-week dexamethasone in treatment on patients with multiple myeloma (MM). Methods 42 patients with MM were diagnosed in our hospital.The VAD regimen contained one-week dexamethasone was vincristine 0.4 mg/d, adramycin 10 mg/d and dexamethasone 40 mg/d for 4 days. At the same time, thalidomide is given 150-200 mg/d. Results The overall response rate (ORR) is 80.95%. Before and after treatment, all the detection indicators in MM patients were statistically different (P＜0.001). There is small adverse reactions. Conclusion Low dose thalidomide combined with VAD regimen contained one-week dexamethasone in treatment of MM has more merits such as less side effects, good tolerance, convenient administration and significant curative effects, so it should be recommended in clinical works.

Objective To explore the feasibility of using the quantitative reference extract of ginkgo leaf total lactones instead of single component reference for the quantitative assay of Ginkgo Folium.Methods HPLC-ELSD method was performed by using a Diamonsil C18 column (250 mm×4.6 mm,5 μm) with methanol-water as the mobile phase at the gradient elution mode.Flow rate was 1.0 mL·min-1.The parameters of ELSD detector were as follows,the drifit tube temperature was 105 ℃,and the flow rate of nitrogen(N2) was 3 L·min-1.Results The linear ranges of ginkgolide A,ginkgolide B,ginkgolide C,and bilobalide were 0.735-5.879 μg (r=0.999 6),0.404-6.060 μg (r=0.999 6),0.296-4.439 μg (r=0.999 6),and 1.001-6.006 μg (r=0.999 7),respectively.The recoveries and RSD of the four components were 95.6% (4.0%),97.3% (4.5%),99.3% (5.0%),and 100.4% (2.1%),respectively.Conclusion The quantitative reference extract of ginkgo leaf total lactones can be used as the substitute for the determination of terpene lactones.

Objective To explore the relationship between the gait behavior changes and cognitive function in APP/PS1 transgenic mice .Methods 16 APP/PS1 transgenic mice were divided into model group and Huperzine A group, C57/BL6J mice with the same age were chosed as control group .After a 150 days consecutive treatment , Morris water maze(MWM) was used to detect the learning and memory ability and Gait analysis system (GAS-2) was used to detect the gait behavior after the treatment when the mice were 8-month-old.Results The escape latency of the model group was significantly higher than that of the control group ( P <0.05 ) , the time spended in the target quadrant , swimming distance in the target quadrant significantly lower than that of the control group ( P <0.05 ) , the first time passing through the platform prolonged significantly than the control group (P <0.05), and the number of passing though the platform reduced significantly than the control group (P <0.05).In the gait behavior experiments , compared with the control group, the average walking speed of the model group reduced significantly (P <0.05), the average walking cycle, the absolute average body angle and lateral movement increased significantly (P <0.05);The percentage of support time in a walking cycle of the left and right foot increased significantly (P <0.05).Accordingly, the percentage of swing time in a walking cycle of the left and right foot reduced significantly (P <0.05).The propulsion index of the left hand , right hand, right foot increased significantly ( P <0.05), and then the braking index of the above three feet decreased significantly ( P <0.05) .Huperzine A can improve the cognitive function , rectify the changes in the gait behavior .The two behavioral relevance shows that cognitive function and the front two feet braking , propulsion index have a high correlation index (correlation coefficients were -0.433, -0.379, P values were 0.039,0.079), the others were not. Conclusion APP/PS1 transgenic mice of 8-months-old have a remarkable impairment of learning and memory ability and disorder of gait behavior , and these two behaviors have a correlation in some extent .

Disability is one of the common geriatric syndromes.Researches addressing elderly disability are still in starting stages.Both opportunities and challenges occur in this field.In this review,we summarized the definition and diagnostic criteria popular assessment tools for the diagnosis of elderly disability,the disease burden and relevant studies regarding elderly disability.

Objective To analyze the influencing factors of quality of life of patients with occupational pneumoconiosis in Methods Chongqing using a structural equation model. A total of 1 181 pneumoconiosis patients in Chongqing were selected - as the research subjects using convenient sampling method. The Chinese version of the SF 36 Scale was used to investigate the quality of life of the patients. Amos 24.0 software was used to construct a structural equation model reflecting the quality of life of Results ， （ ），（ ） （ ）， patients. The scores of physical health mental health and quality of life were 61.7±18.5 68.7±20.5 and 64.7±16.8 respectively. The scores of the eight dimensions of the quality of life of the research subjects were lower than that of the （ P ） Chongqing City municipal and national norms all <0.01 . Multiple linear regression results showed that personal monthly ， ， ， income pneumoconiosis stage complications employment status and smoking were the influencing factors of the quality of life （ P ） ， of pneumoconiosis patients all <0.05 . The results of the structural equation model showed that the stage of pneumoconiosis ， ， complications personal monthly income and pulmonary rehabilitation treatment could directly affect the physical and mental （ P ） ， health of the research subjects all <0.05 . In addition the stage of pneumoconiosis and complications indirectly affected the （ P ）Conclusion physical and mental health of patients by affecting pulmonary rehabilitation treatment all <0.05 . The structural equation model can be used to verify the influencing factors of the quality of life of patients with occupational pneumoconiosis. ， Complications personal monthly income and pneumoconiosis stage are the main factors affecting the quality of life of patients with occupational pneumoconiosis.