Objective To develop a method of adult porcine pancreatic islet isolation.Methods The tails of adult porcine pancreas were perfused through the pancreatic duct with 0.1% cold collagenase(type Ⅺ) and incubated at 38.5 ℃.The digested tissue was dispersed in 4 ℃ Hanks balanced salt solution(HBSS).The tissue suspensions were filtered through a 600 ?m mesh.The residual tissue was resuspended in cold HBSS,and put in the Ricordi’s chamber and shaken for 5 minutes,then filtered again.The isolated islets were divided into three groups: control group( n =14),Pefabloc(trypsin inhibitor, n =8) group and FOY(trypsin inhibitor, n =5) group.The collagenase solution of the Pefabloc and FOY group was supplemented with 1.0 mmol/L Pefabloc and FOY respectively. Results The islet yields of the Pefabloc group and FOY group 〔(11 848?3 530) islet/g pancreas and (14 496?3 693) islet/g pancreas〕 were significantly higher than that of the control group 〔(8 505?3 349) islet/g pancreas〕, P

Bone metastases in advanced lung cancer patients are common and affect the life quality seriously. So early diagnosis and appropriate treatments are important. The methods of diagnosis include X-ray, CT, MRI, ECT and PET, which are different in sensitivity and specificity. The gene microarray technique is another advance for detecting micrometastasis. The gene analysis also can help understanding the biological characteristics of the primary tumor. The treatment of bone metastases in lung cancer is various, including systemic and local therapeutic means. Surgery is often used to prevent or treat the compressions and bone fractures. Radiotherapy is one of the most effective methods of pain relief. Diphosphonate provides a new convenient and effective way of alleviative treatment. In addition, supportive and symptomatic treatments should be keep in mind.

OBJECTIVE:To provide reference for continuous improvement of drug counseling and pharmaceutical care. METH-ODS:220 cases of drug counseling answered by inpatient general pharmacist were collected from our hospital during 2012-2013, and the contents of drug counseling were classified and analyzed. RESULTS:Among consultants,73.18% of them were physi-cians;the type of drug involved mainly were antibiotics,accounting for 42.73%;main contents of drug counseling included drug selection,usage and dosage,accounting for 47.27%. CONCLUSIONS:Pharmacists provide drug counseling for physicians and nurses,answer various questions about usage,and guide rational drug use so as to enhance the communication between pharma-cists and physicians,nurses,and promote rational drug use in the clinic.

Objective To construct the bicistronic retroviral vectors containing GFP gene and human survivin gene(wildtype and T34A mutant). Methods Survivin fragments were amplified from eukaryotic expression vectors by PCR, then cloned into pMIG retroviral vectors. The constructed recombinant was identified by double digestion with restriction enzymes. The constructed retroviral vectors was transfected into Phoenix E package cells under mediation of liposome. The expression of mRNA was examined by RT-PCR and the transcription of gene was determined by FACS analysis of GFP. Results The target fragments were successfully bound to pMIG vector. After the vectors were transfected into NIH3T3 cells, the expression of mRNA was confirmed by RT-PCR and the expression of GFP was confirmed by FACS analysis. Conclusion Human survivin gene wildtype and T34A mutant retroviral vectors have been constructed successfully, which can provide strong molecular tools for further studies and a novel approach for selective cancer gene therapy.

Objective　To analysis the relationship between the multipathogens infection and unhealthy pregnancy.Methods　All patients was selectively divided into three groups:group A including 106 patients with unhealthy pregnancy,group B:55 gravidas with normal induced abortion,group C:55 normal parturient with term labor.The ten kinds of pathogens from abortion tissues and placentas were tested by PCR method respectively.Anti-pathogen IgM antibodies were detected by ELISA in the blood specimens.Results　The distribution status of the pathogens were as followed:Human parvovirus B19(B19V 23.6%)＞Cytomegalovirus(CMV,16.0%)＞Toxoplasma (TOX，15.1%)＞Rubella virus(RV，10.4%)＞Ureaplasma urealyticum(9.4%)＞Coxsackie(7.5%)＞Herpes simplex virus(HSV,6.6%)＞Adenovirus,Eprstan Barr virus(3.8%)＞Chlamydia trachomatic(CT 2.8%) in the group A.The positive rates of the pathogens infection in both group C and group B were lower.The positive rates of B19V,CMV,RV,HSV,TOX were significant difference(χ2 test,P＝0.000～P＝0.031) among the three groups,hovever others were not(χ2 test,P＝0.121～P＝0.724).Conclusions　The pathogen infection is an important factor to occur the unhealthy pregnancy,the major pathogens of unhealthy pregnancy are B19V，CMV，TOX，RV，HSV.These pathogens infection should be detected timely in gestational period.

Objective:To assess the clinical outcome of autologous fat injection in treatment of hemifacial atrophy,so as to search for an ideal treatment of hemifacial atrophy.Methods: Autologous fat granules were obtained by tumescent liposuction.After purification,the fat granules were injected in a multi-strata,multi-tunnel,and multi-spot manner to repair facial introcession abnormality.Patients whose introcession was not improved after the stabilization of the fillings were re-injected with autologous fat granules twice or more until satisfactory outcomes were achieved.Results: All the 7 patients in this study obtained satisfactory outcomes after fat injection and basically reached symmetry.The follow-up result was good after 2 years.Conclusion: Local injection transplantation of autologous fat granules is effective for the treatment of facial depression and hemifacial atrophy,with slight trauma,little complication,and satisfactory outcome.

Objective:To investigate the anatomic characteristics of the pudendal-thigh flap and provide anatomic basis for clinical design and application. Methods: The perineal regions of 15 male cadavers were anatomized. The flap was designed on the cadavers and the blood vessels/nerves contained in the flap were studied. Based on the above anatomic findings, we designed the flaps in our operation. The axial arteries were isolated according to the designing and the tissue adjacent to the vessel pedicles was preserved. The flaps were transferred to the recipient site. The incision of donor site was directly sutured; skin graft was transplanted when the incision could not be directly sutured. Results: The perineal regions had rich and constant blood supply and had stable nerve innervation. The blood supplies of the perineal region included external pudendal artery, anterior cutaneous branches of obturator artery, lateral branch of posterior labial or scrotum arteries. The location of the above vessels was comparatively constant, especially external pudendal arteries and posterior scrotum arteries. Three groups of blood vessels went through the lower, middle and upper parts, and there were 4 groups of major nerves responsible for the innervation of the pudendal region. We have used 23 cases of pudenal-thigh flaps, with the largest flap being 17 cm X 9 cm and the smallest being 8 cm X 5 cm, and satisfactory outcomes were achieved in all the cases. Conclusion: The pudenal thigh flap has rich blood supply, convenient incision, and covert donor site. It has satisfactory outcome (morphologic and sensory) in repairing and reconstructing male perineal region and is worth popularizing.

Objective To study relationship between blood-glucose change and prognosis of the patients after severe traumatism.Methods 70 cases severe traumatic emergency patients selected tested the level of blood-glucose instantly when they presented themselves to the clinic after injury,used saccharimether and blood-glucose test paper provided by the Johnson Limited Corporation,and 30 slight traumatic emergency patients at the same time were selected as contrel.Result The blood-glucose after severe traumatic were markedly higher than that of slight traumatic group,which have significance(P

Objective To study the inhibitory effects of tea flower extract (TFE) on α-glucosidase and glucose intestinal absorption. Methods Three different postprandial hyperglycemia models (2 g/kg glucose, 4 g/kg sucrose, and 6 g/kg starch) were used, with 8 mice in each group. Oral administration of 150 or 300 mg/kg of TFE, 6.25 mg/kg of acarbose, or water was performed on mice 1 day and 30 mins before the oral administration of 2 g/kg glucose, 4 g/kg sucrose, and 6 g/kg starch at 10 ml/kg of body weight. Blood glucose levels were analyzed chronologically to evaluate the effect of TFE. In vitro studies were also performed to study the inhibitory effects of TFE on α-glucosidase and small intestinal mucosa glycosidase.

Objective To study a-synuclein(SNCA)rs356200 and rs356165 loci polymorphism in the patients with Parkin son's disease(PD)and to investigate the correlation between SNCA gene polymorphism and PD cognitive impairment.Methods One hundred and twenty cases of PD in our hospital from June 2014 to December 2015 and 120 heaalthy people were selected as the research subjects.The PD cases served as the PD group and 120 healthy subjects as control group.The PD cases were also divided into the cognitive impairment group and non-cognitive impairment group according to the cognitive impairment.The rs356200 and rs356165 allele frequencies and genotype in each group were studied.Results The rs356200 allele frequency had statistical difference between the PD group and control group(P＜0.05),the rs356200 G/G,A/A and A/G genotype had statistical difference between the PD group and control group(P＜0.05).The rs356165 allele frequency had no statistical difference between the PD group and the control group(P＞0.05),the rs356165 G/G,A/A and A/G genotype had statistical difference between the PD group and control group(P＜0.05).The rs356200 and rs356165 allele frequencies and genotype had no statistical difference between the cognitive impairmentgroup and non-cognitive impairment group(P＞0.05).Conclusion The morbidity of PD is related to SNCA gene rs356200 and rs356165 sites polymorphism.PD cognitive dysfunction has no significant correlation with SNCA gene rs356200 and rs356165 loci polymorphism.