Suppression subtractive hybridization(SSH) is a good and highly effective method for gene cloning,which posses some character,such as convenient operation,high specificity,low background,good replication etc,and it can separate lower specific fragment.This article principally introduced its main technical points,strength and shortcomings and improvement of SSH,and also reviewed its applications in detecting the effects of radiation,heat,hypoxia,coldness and other environmental factors on gene expression.

Objective To investigate the feasibility and safety of laparoscopic surgery of giant hysteromyoma.Methods A total of 56 cases of giant hysteromyoma(at least 1 lesion ≥ 9 cm in diameter) underwent laparoscopic operations from January 2003 to August 2006 in this hospital.There were 30 cases of laparoscopic myomectomy(LM),10 cases of laparoscopic-assisted vaginal hysterectomy(LAVH),1 case of total laparoscopic hysterectomy(TLH),and 15 cases of laparoscopic supracervical hysterectomy(LSH).Results All of the operations were successfully performed under laparoscope.No conversion to open surgery was needed.The operation time was 40～100 min(mean,52 min),and the hemorrhage volume was 10～100 ml(mean,45 ml).Postoperatively,ureterovaginal fistula was observed in 1 case,which was cured by a re-operation of laparoscopic ureteroneocystostomy.Conclusions Laparoscopic operation for giant hysteromyoma is safe and feasible.

Objective To evaluate efficacy and safety of laparoscopic surgery for ectopic pregnancy with hemorrhagic shock.Methods One hundred and thirty women with hemorrhagic shock due to ectopic pregnancy underwent laparoscopic surgery from January 2003 to August 2006,including salpingectomy in 80 cases,partial ovariotomy in 8 cases,uterus wedge resection in 15 cases,salpingotomy in 19 cases,and tubal lesion clearance in 8 cases.Results Laparoscopic surgery was successfully performed in all the cases.Intra-abdominal blood loss was 1000 to 3500 ml,with an average of 1500 ml.The operation time was 30-60 minutes(mean:40 minutes).No postoperative complication occurred.Follow-up was conducted in 106 cases,once in a week in the first month after discharge.Serial ?-hCG levels decreased gradually and reached the normal level 1-2 weeks after operation.Persistent ectopic pregnancy was not found in the studied cases.Conclusions Laparoscopic surgery is a safe and effective treatment for hemorrhagic shock due to ectopic pregnancy,given the skill of an experienced laparoscopist with the aid of optimal anesthesia and advanced cardiovascular monitoring.

Objective To investigate the diagnostic value and therapeutic effect of laparoscopy combined with hysteroscopy in patients with tubal infertility.Methods A total of 133 patients with tubal infertility were treated by laparoscopy combined with hysteroscopy in our hospital from September 2004 to December 2007.All the cases were diagnosed by hysterosalpingography(HSG) and hysteroscopy.Results Totally 225 fallopian tubes were examined in the 133 patients,among which 180 tubes were completely occluded,57 were partially occluded,and 18 were patent.After the surgeries,211 tubes wee patent,24 were partially blocked,while 20 were still occluded.The success rate of the operations was 81.4%(193/237).Follow-up was achieved in 92 patients for 4 to 18 months,in the cases,30 were pregnant during the period(pregnancy rate: 32.6%).Of the 30 cases,2 women had tubal pregnancy;22 women were pregnant in 6 months after the treatments,and the other 8 were pregnant in 6-12 months after the operations.In this group of patients,the pregnancy rate in the women with both patent fallopian tubes was 26.1%(24/92),and in those with single patent fallopian tube was 6.5%(6/92).The pregnancy rate in patients with simple tubal infertility was 22.8%(21/92),while that of those who were complicated with tubal diseases was 9.8%(9/92).Conclusion Laparoscopy combined with hysteroscopy can accurately diagnose tubal infertility,and improve the therapeutic effect.

Objective To investigate the effectiveness and safety of endovascular coiling and microsurgical clipping for ruptured intracranial aneurysms. Methods Patients w ith ruptured intracranial aneurysm treated w ith endovascular coiling or microsurgical clipping w ere enrol ed retrospectively. The demography, baseline clinical data, outcome, and complications in patients received endovascular coiling and microsurgical clipping w ere compared. Results A total of 85 patients w ith ruptured intracranial aneurysm were enroled, including 40 were treated with microsurgical clipping (surgical clipping group) and 45 were treated w ith endovascular coiling (endovascular coiling group). There w ere no significant differences in the proportions of the patients in male (37.5%vs.40.0%; χ2 =0.056, P=0.813), hypertension (30.0%vs. 33.3%; χ2 =0.109, P=0.742 ), smoking ( 50.0%vs.48.9%; χ2 =0.010, P=0.918 ), drinking (45.0%vs.46.7%; χ2 =0.024, P=0.878), aneurysm site (anterior communicating artery: 50.0%vs. 48.9%;posterior communicating artery:35.0%vs.33.3%; middle cerebral artery:10.0 %vs.11.1%;vertebral artery: 5.0%vs.6.7%; al P>0.05), aneurysm maximum diameter < 10 mm (80.0%vs. 77.8%;χ2 =0.063, P=0.802), Hunt-Hess grade 1-2 (55.0%vs.57.8%; χ2 =0.066, P=0.797), Fisher grade 1-2 ( 60.0%vs.57.8%; χ2 =0.043, P=0.835 ), and time from onset to treatment < 72 h (62.5%vs.64.4%; χ2 =0.035, P=0.853) in the surgical clipping group and endovascular coiling group. There w ere no significant differences in the complete occlusion rate of aneurysms ( 97.5%vs.91.1%;P=0.364) and the good outcome rate (65.0%vs.68.9%; χ2 =0.145, P=0.703) betw een the surgical clipping group and the endovascular coiling group. No patients died in the surgical clipping group and 1 patient died in the endovascular coiling group, and there w as no significant difference ( P=1.000). One patient (2.5%) had cerebral infarction in the surgical clipping group and no patients had cerebral infarction in the endovascular coiling group, and there w as no significant difference ( P=0.471). Conclusions The efficacy and safety of microsurgical clipping are the same as those of endovascular coiling for ruptured intracranial aneurysms.

Objective To compare the efficacy, complications, safety and prognosis of the minimally invasive puncture approach and key hole in the treatment of hypertensive cerebral hematoma.Methods A totol of 68 patients with hypertensive cerebral hematoma confirmed by CT from April 2012 to October 2013 in Nongken Sanya Hospital were randomly divided into key hole evacuation group(n=32) and minimally invasive puncture group (n =36).Comparisons were made between the two surgical methods in the operative time, postoperative complications, the fatality and the postoperative re-haemorrhagia rate, neurological function deficit score also been observed and evaluated in the 1 st,2nd and 4th weeks after surgery.Results The NFDS scores of the two groups both decreased in the 1st week after surgery,but compare with preoperative the difference was not statistically significant (P ＞ 0.05).In the 2nd weeks and 4th weeks after surgery, NFDS scores further decreased in both group,and there was statistically significant compare with preoperative(the key hole evacuation group : (26.2±4.5) vs.(17.8 ± 3.6) vs.(44.1 ± 5.4) scores;the minimally invasive puncture group: (22.1 ± ±3.7) vs.(15.4±2.8) vs.(43.9±6.2)scores;P＜0.05) ,but during the same period there was no significant difference between the two groups with NFDS scores(P＞0.05).The rebleeding rate of the minimally invasive puncture group was significantly lower than the key hole evacuation group (4.08％ vs.16.33％, x2=6.56, P＜0.05).There was no significant difference in mortality rate and long term total effect between two groups (P＞0.05).Conclusion Although both key hole and minimally invasive puncture are effective measures for treatment of hypertensive cerebral hemorrhage, but minimally invasive puncture with less trauma, definite curative effect and higher security advantages in clinical.

ObjectiveTo investigate the effect of vitamin D supplementation on the outcome of acute ischemic stroke in young patients with vitamin D deficiency.MethodsThe prospective controlled study was used to select the consecutive young patients with acute ischemic stroke.Vitamin D deficiency was defined as 25-hydroxyvitamin D (25(OH)D) ≤50 nmol/L.The Patients with vitamin D deficiency were randomly divided into an intervention group and a routine treatment group according to the random number table method.Routine treatment group didn't receive the drug intervention for vitamin D deficiency, and the intervention group received daily oral alfacalcidol 0.5 μg.After 1 year of treatment, the 25(OH)D levels were examined again;the adverse reactions during the drug treatment were monitored;the modified Rankin Scale (mRS) was used to evaluate the functional outcome and 0-2 was defined as good outcome.ResultsThere are 94 patients (53.41%) with vitamin D deficiency among 176 young patients with acute ischemic stroke.They were randomly divided into either an intervention group or a routine treatment group (n=47 in each group).At the end of the follow-up, the good outcome rate (82.98% vs.63.83%;χ2=4.414, P=0.036) and serum 25(OH)D level (85.83±10.53 nmol/L vs.39.10±11.18 nmol/L;t=20.860, P<0.001) in the intervention group were significantly higher than those in the routine treatment group.During the follow-up period, there was no loss to follow-up or death events in both groups.Only 2 cases of nausea and 1 case of dizziness were observed, and the incidence of adverse reaction was 6.38% in the intervention group.ConclusionsVitamin D supplement can increase the vitamin D levels and improve functional outcome in young patients with acute ischemic stroke and vitamin D deficiency.

Mild cognitive impairment is a transitional stage between normal aging and dementia. It is important in terms of recognizing memory loss in older people as well as identifying a group of individuals at high risk of developing dementia and who may benefit from preventive strategies. Ginkgo biloba extract has been shown to possess polyvalent properties, such as anti-oxidation, anti-apoptosis and anti-inflammation. Ginkgo biloba extract appears to have a neuroprotective effect against neurodegenerative diseases.

Aim To provide practical method for screening human carbonic anhydrase Ⅱ(hCA Ⅱ) inhibitors in drug discovery.Methods hCA Ⅱ protein was obtained from induced BL21(DE3) E.coli containing plasmid pET-28b-hCA Ⅱ.The hCA Ⅱ activity was detected under pH 7.6 and 25℃ by its esterase activity which could decompose PNPA to increase the photoabsorption at 348 nm. After the assay conditions were finally selected, 35 new compounds were tested.Results A practical method for screening hCA Ⅱ inhibitors was successfully constituted by using recombinant hCA Ⅱ protein expressed in E.coli as the source of hCA Ⅱ enzyme.10 new compounds had better inhibitory effect and 9 new compounds had the same inhibitory effect on hCA Ⅱ compared with acetazolamide.Conclusions The hCA II inhibitor screening technique constituted in this work possesses advantages of being reliable, rapid, and practical. 19 new compounds are worth further research for developing high efficiency and low side effect drugs used for high-altitude illness.

ObjectiveTo discuss the clinical value of serum CA125 and endometrial antibody (EMAb) for the diagnosis of endometriosis.Methods216 patients were determined by the presences of CA125 and EMAb before operation.ResultsAll cases were diagnosed by pathology after operation. CA125 positive rate in the endometriosis group was 58.3% and that in the control group was 12.5%. The difference between two groups was significant (P<0.01).EMAb positive rate in the endometriosis group was 31.3% and that in the control group was 14.3%. The difference between two groups was also significant (P<0.01). When determining CA125 alone to diagnose endometriosis, the sensitivity rate was 58.3% and specificity rate was 87.5%. If determining EMAb alone to diagnose endometriosis, the sensitivity rate was 31.3% and specificity rate was 85.7%. When one of them was used as diagnostic criterion, the sensitivity and specificity were 64.6% and 73.2% respectively. If combining use of both CA125 and EMAb as diagnostic criterion, the sensitivity and specificity were 25.0% and 100% respectively.Conclusions The determination of serum CA125 or EMAb levels is helpful for the qualitative diagnosis of endometriosis, especially using them combined, the diagnostic accuracy may be enhanced.