BACKGROUND: Construction of lentivirus vector has the advantages of simplicity,it is regard as the most effective and successful method in transgene therapy. OBJECTIVE: To clone the metastasis suppressor gene KiSS-1 from human normal placenta tissue and construct its lentivirus vector. DESIGN,TIME AND SETTING: The open experiment was performed at the Laboratory of Fujian Normal University from September 2006 to December 2007. MATERIAL: The pNL-IRES2-EGFP vector was conservated by the Laboratory of Fujian Normal University. METHODS: Total RNA was extracted from human placenta tissue. The opening reading frame cDNA of KiSS-1 was isolated by using RT-PCR,and cloned into its lentiviral vector pNL-IRES2-EGFP to construct expression plasmid pNL-IRES2-EGFP-KiSS-1. MAIN OUTCOME MEASURES: Clone objective gene fragment of KiSS-1,restriction enzyme digestion and gene sequencing of the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 were observed in the study. RESULTS: The nucleotide sequence isolated from the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 was confirmed the same as expected by restriction enzyme digestion and gene sequencing. CONCLUSION: The recombinant plasmid pNL-IRES2-EGFP-KiSS-1 has been constructed successfully.

To evaluate the application of wavefront aberration in diagnosis and operation of retinal detachment.Totally 17 rhegmatogenous retinal detachment patients(17 eyes),who received outer scleral cryocoagulation,silicon cushion and(or)intravitreous injection of gas,were selected from July 2004 to February 2005 at First Hospital Affiliated to Dalian Medical University.These cases were all retinal detachment in one eye.Of the 17 patients,10 were diagnosed as having myopic eye retinal detachment and 7 were non-myopic eye retinal detachment.The wavefront aberrations of these patients were checked with wavefront aberrometer produced by German Wavelight Laser Technology AG Company before operation,1 and 3 months after operation,and compared with the other eyes.The results showed that ①Root mean square(RMS)1,RMS3-RMS6 and root of mean square of higher aberration(RMSH)were significantly higher than those in the control group before operation(P 0.05).②RMS1,RMS3 and RMSH in the myopic retinal detachment eyes were lower than those in the non-myopic retinal detachment eyes before operation(P

Objective To establish the HPLC fingerprint analysis method for the quality control of Flos Daturae. Methods Column:Zorbax C18 (4.6 mm?250 mm, 5 ?m);Mobile phase:0.07 mol/L NaPO4 (adjust pH to 6.0 with phosphatic acid, including 17.5 mmol Soldium dodecyi Sulphate)-Acetotrile (2∶1). Flow rate was 0.8 mL/min and detection wave length was 216 nm. Results The repetition and precision of the method was good. There was obvious difference between South and North Flos Daturae based on the results of cluster and similarity analysis. Conclusion The method can control the quality of Flos Daturae effectively.

Aim To explore the mechanisms of Curcumin which plays a role of protecting cells through regulating the activity of GSK-3?.Methods Plasmid APPswe and BACE1-mychis were transiently co-transfected into SHSY5Y cells.The cells were treated with Curcumin and then were collected to determine the mRNA and protein levels of GSK-3?,?-catenin and CyclinD1 by RT-PCR and Western blot. Immuno-fluoresecnt staining was used to detect the location and expression of ?-catenin and GSK-3?, and ELISA was for the generation of A?.Results RT-PCR and Western blot results showed that in the cells, Curcumin inhibited the expression of GSK-3? mRNA and protein, and promoted the expression of GSK-3?-Ser9 protein (P

Fluid therapy plays a quite important role in peri-operation period,however,conventional measurements are not sensitive to hypovolemia.Goal directed fluid therapy( GDT),which is based on balance of oxygen delivery and consumption,can achieve the aim of fluid therapy more efficiently.Many RCTs have proven that GDT could reduce morbidity,shorten hospital time of high risk surgical patients,therefore,it is a beneficial work.

OBJECTIVE:To understand the availability and affordability of essential medicines in Nanjing after implementing national essential medicine system,and to provide evidence for formulating and completing related policy. METHODS:With inves-tigation subjects of 6 classes of diseases with high incidence,WHO/HAI standard survey method was adopted to select 24 essential medicines and investigate and evaluate the availability and affordability of them in governmental health institutions and social retail pharmacies in Nanjing. RESULTS:Availability of essential medicines in community health service centers in Nanjing was lower than governmental hospitals and social retail pharmacies,and availability of lowest price generic drugs (LPGs) was much higher than original brand drugs (OBs);affordability was totally good,however,there were large gap in the affordability between OBs and LPGs,the affordability of OBs was relatively poor. CONCLUSIONS:The national essential medicine system has made some progress in Nanjing,but worse than expected. It is suggested to update and adjust the essential medicine list based on the clinical medication requirement,guarantee the equipment of essential medicines in medical and health institutions,popularize the knowl-edge about essential medicines,properly adjust the price of original brand drugs and ease the treatment burden.

Objective To explore the expression of mismatch repair gene (MMR) (MLH1,MSH2,PMS2 and MSH6) in endometrioid adenocarcinoma,and to analyze its clinical pathological significance.Methods Inmunohistochemical EnVision method was used to detect the expressions of DNA mismatch repair proteins (MLH1,MSH2,PMS2 and MSH6) in 101 cases of endometrial adenocarcinoma.The phenotypes of tumor microsatellite instability (MSI) and microsatellite stable were determined,and its relationship with onset age,differentiation,depth of myometrial invasion,lymphatic metastasis and prognosis of tumor were analyzed.Results There were 67 cases of hysterectomy and 34 cases of endometrial biopsy in 101 cases of endometrial adenocarcinoma.52 cases of surgical specimens and 14 cases of biopsy specimens were included in 66 cases that had complete follow-up data.Rates of negative expression were as follows:MLH1 31.6 ％ (32/101),MSH228.7 ％ (29/101),PMS2 16.8 ％ (17/101),MSH6 8.9 ％ (9/101).MSI phenotype 53.5 ％ (54/101),MSS phenotype 46.5 ％ (47/101).Univariate analysis showed that prognoses of MSI-L&MSS group (34 cases) were better than those of MSI-H group (18 cases) in hysterectomy organization (P =0.041).The differences between depth of myometrial invasion,degree of differentiation,age and prognosis of endometrial adenocarcinoma were statistically significant (P ＜0.05).Cox multivariate analysis showed that differences between depth of myometrial invasion,age and prognosis of endometrial adenocarcinoma were statistically significant (P =0.034,P =0.009),while there was no significant difference between MSI-L&MSS group and MSI-H group in prognosis (P ＞ 0.05).Conclusions MSI is one of the molecular events that occur in the endometrioid adenocarcinoma.Age and depth of tumor invasion are independent prognostic factors of endometrioid adenocarcinoma.In view of inconsistency between univariate analysis and multivariate analysis,whether MSI can be used as deterministic accordance for endometrioid adenocarcinoma prognostic evaluation requires further verification.

Objective To evaluate the effect of dexmedetomidine on baroreflex in the patients.Methods Forty-five ASA physical status Ⅰ or Ⅱ patients,aged 20-60 yr,with body mass index 18-24 kg/m2,scheduled for elective partial thyroidectomy or nasal polypectomy under general anesthesia,were randomly divided into 3 groups (n =15 each) using a random number table:control group (group C),low-dose dexmedetomidine group (group LD) and medium-dose dexmedetomidine group (group MD).Dexmedetomidine 0.5 μg/kg was injected intravenously followed by infusion at 0.2 μg· kg-1 · h-1 in group LD.Dexmedetomidine 1.0 μg/kg was injected intravenously followed by infusion at 0.5 μg· kg-1 · h-1 in group MD.After 30 min of dexmedetomidine infusion,a modified Oxford pharmacologic technique was used for evaluating baroreflex sensitivity.Results There was no significant difference in baroreflex sensitivity between the three groups.Conclusion Dexemedtomidine exerts no effect on baroreflex in the patients.

Objective To investigate the long-time effects of early recurrence treated by aptos thread after skin grafting of lower eyelid ectropion.Methods Eight eyelids of 6 patients with early recurrence after skin grafting of lower eyelid ectropion were reviewed.They were operated between January 2009 and January 2013.Surgeries were done under local anesthesia.Aptos thread was fixed on the deep temporal fascia above the outer canthus and implanted under the orbicularis oculi muscle.Then aptos thread was fixed with the periorbit of inner canthus.Results 8 eyes of 6 patients were followed for 6～12 months.All the patients were satisfied with the good closure function,ideal cosmetic outcome and non-recurrences of lower eyelids extropion.Conclusions The technique of deep temporal fascia and periorbit fixed by aptos thread is an effective method for early recurrence after skin grafting of lower eyelid ectropion.It would be helpful to obtain a lasting post-operative effects.

Objective To explore a technique to correct severe congenital nipple inversion,which has preferable clinical curative effect and lower recurrence rate.Methods 16 patients with bilateral severe congenital nipple inversion were included in the study.The rhomboid skin incision and triangular dermal breast compound tissue flap was designed in two directions.One compound tissue flap was pulled out through the tunnel under the nipple to the opposite side and sutured at the base of the other flap.The cross-fixation was accomplished to support the nipple tissue.The shortened lactiferous ducts and fibrous bands beneath the nipples were released.The nipple neck was tightened by suturing the shorter diagonal of rhomboid skin incision.Results No complications,such as infection,hematoma and necrosis,were found in all cases.All nipples manifested satisfactory aesthetic projection without recurrence after 6-24 months follow-up.Conclusions In the treatment of severe congenital nipple inversion,the technique of the nipple suspension using two triangular dermal breast compound tissue flaps could provide a strong suspension system with less recurrence rate.And the nipple neck could be tightened by suturing the shorter diagonal of rhomboid skin incision in the meantime.The nipple will manifest satisfactory aesthetic projection.