Objective To study an improved automated method for isolation and purification of large amounts of human pancreatic islets from large mammals,and try to create conditions for preparation and isolation of large amounts of human islets.Methods An improved automated system was used to isolate and purify panreatic islets of sogs.Under the general anaesthesia(GA) condition,the pancreas of the dogs was via in situ vascular perfused using cold HC-A solution and then removed en bloc with duodenum and spleen.Then they were placed in cold UW aolutian.Intraductal collagenase-Ⅴ and pefabloc(4 ℃) was delivered with controlled perfusion.lslets were dissociated in system of Ricordi Chamber and purified islets separated with Ficoll density gradient centrifugation under controlled temperature.Digestion time,islets remaining trapped in exocrine tissue,final islet purity,insulin and C-pep secretory activity,and islet recovery were observed.The purified islets were observed under light microscope and electronic microscope after 24 h culture.Results The digestion time rate was(25.0?6.0) min,islets remaining trapped in exocrine tissue was(9.4?2.4)%,final islet purify rate was(89.7?3.5)%,islet recovery after digestion was(17.2?3.6)?104 IEQ/pancreas.Islet recovery after purification was(8.3?2.0)?104IEQ/pancreas.Insulin and C-pep secretory activity of the purified islets and their ultrastructure after 24 h culture were ideal.Conclusions Our improved automated method and facilities for isolation of canine pancreatic islets are reliable,The morphology and function of the procured islets are excellent and they can foreseeably be used for large-scale preparation of huma islets for clinical use.

Actins ; metabolism ; Aged ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Fasciitis ; metabolism ; pathology ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratin-5 ; metabolism ; Mastectomy, Modified Radical ; Neoplasms, Muscle Tissue ; metabolism ; pathology

OBJECTIVETo study the effect of Shenqi Fuzheng Injection (SQFZI) on erythrocyte immune function in patients after cardiopulmonary bypass (CPB).

METHODSTwenty patients ready for receiving CPB were randomly assigned to two groups, 10 in each group. Patients in the SQFZI group were treated with 250 mL SQFZI via intravenous dripping starting from 5 days before operation and after ending CPB once a day. Peripheral venous blood samples of all patients were collected at the time points of before anesthesia, and 6 h, 24 h, 72 h and 7 days after CPB, which were anticoagulated with heparin for dynamically detecting the changes of RBC-C3bRR and RBC-ICR.

RESULTSRBC-C3bRR and RBC-ICR in the SQFZI group were significantly higher at 6 h, 24 h, 72 h, and day 7 after operation than those in the control group respectively at the corresponding time points, and they restored to the normal levels at day 7, suggesting the erythrocyte immune function after CPB in the SQFZI group was superior to that in the control group (P < 0.05).

CONCLUSIONApplication of SQFZI in the peri-operative period can significantly improve the hypo-immunological function of erythrocyte caused by CPB, and promote the recovery of erythrocyte immunity.

Adult ; Aged ; Cardiopulmonary Bypass ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Erythrocytes ; drug effects ; immunology ; Female ; Heart Valve Diseases ; blood ; drug therapy ; surgery ; Heart Valve Prosthesis Implantation ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Phytotherapy ; Treatment Outcome

Background Bulbar conjunctiva tissue appears to be thinning,elasticity declined,tension reduced and fascia atrophied in conjunctivochalasis.Histopathological examination of conjunctivochalasis shows decrease of elastic fibers and melt of collagen fibers.But there are fewer studies on the ultrastructure of conjunctiva of conjunctivochalasis up to now.Objective This study was to investigate the ultrastructure change of conjunctiva tissue in conjunctivochalasis.Methods Five loose conjunctiva samples of conjunctivochalasis and 5 normal conjunctival tissue samples were collected and ultra-microstructure changes of these samples were observed under the transmission electron microscope.Results The number of fibroblasts in conjunctivochalasis lamina was progressively decreased.The shape of fibroblasts was long and fusiform.Somatic synapse was slim.Nucleus-cytoplasm ratio was increased.Disorder,scattered and broken of the collagen fibril were seen,and some areas were dissolved or lacunae.Normal conjunctival fibroblasts were oval or polygonal,with wieners and long somatic synapse,and intercellular matrix was full of collagen fibril and dense arranged fibers.Fibroblasts in fascia of eonjunctivochalasis were linear,and collagen fibril was seriously defected.Fascia fibroblasts of normal bulbar conjunctiva were spindleshaped and bigger than conjunctivochalasis fibroblasts.There were full of collagen fibrils in intercellular matrix.Conclusions The collagen fibril is decreased and fibroblast cells are degenerated in lamina and fascia of conjunctivochalasis.

Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

Objective To analyze the therapeutic effect of noninvasive mechanical ventilation in chronic obstructive pulmonary disease (COPD) patients with respiratory failure.Methods Sixty patients with COPD respiratory failure were randomly divided into two groups.30 cases in the conventional treatment group used conventional comprehensive therapy.30 cases in the nonfinvasive mechanical ventilation group were given noninvasive mechanical ventilation based on conventional treatment.The total effective rate,time to correct the abnormal breathing,hospitalization time were compared between the two groups.Before and after treatment,the arterial blood gas analysis indicators and heart rate,respiratory rate,probability,mortality for ventilator ventilation were compared in the two groups.Results The total effective rate of the noninvasive mechanical ventilation group was 96.67％,which was higher than 73.33％ of the conventional treatment group(x2 =6.405,P ＜ 0.05).The time to correct the abnormal breathing,hospitalization time of noninvasive mechanical ventilation group were (5.51 ± 1.41) d,(9.51 ± 2.13) d,respectively,which were shorter than (9.39 ± 2.56) d,(12.99 ± 3.57) d of the conventional treatment group (t =8.223,9.633,all P ＜ 0.05).Before treatment,the arterial blood gas analysis parameters and heart rate had no statistically significant differences between the two groups (all P ＞ 0.05).After treatment,the frequency of PaCO2,PaO2,heart rate and respiratory rate in the conventional treatment group were (60.45 ± 3.94)mmHg,(60.24 ± 4.12)mmHg,(92.58 ± 0.61)times/min,(22.45 ± 3.14)times/min,respectively,which in the noninvasive mechanical ventilation group were (52.24 ± 1.23) mmHg,(76.24 ± 5.26) mmHg,(83.92 ± 0.62) times/min,(17.26 ± 2.23) times/min,respectively,the differences between the two groups were statistically significant(t =5.133,3.734,5.166,6.360,all P ＜ 0.05).The invasive rate of invasive ventilator and mortality of noninvasive mechanical ventilation group were 3.33％ and 0.00％,respectively,which were lower than 20.00％,13.33％ of the conventional treatment group (x2 =7.214,6.247,all P ＜ 0.05).Conclusion Noninvasive mechanical ventilation is effective in treatment of patients with COPD respiratory failure,which can effectively improve the indicators of arterial blood gas and heart rate and respiratory rate,improve the prognosis of patients,improve the ventilation probability of invasive ventilator.

Objective To describe the characteristics of sexual debut among college students in China to provide appropriate sex education in that population. Methods An anonymous questionnaire survey was conducted among 33 653 college students selected by multi-stage stratified cluster sampling method in 18 provinces and cities of China. Results The percentage of students who had initiated sexual intercourse before age 14 was 1.9%. Overall, male students (2.8%) were significantly more likely to have initiated sexual intercourse before age 14, than female students (1.1%). The percentage of students who had initiated sexual intercourse from age 15 to 18 was 6.4% in males versus 2.1% in females. while the rates of students who had initiated sexual intercourse after age 19 were 6.9% in males and 2.6% in females. The first sexual intercourse was significantly associated with parental educational attainment, geographic location and family structure. Students whose parents were relatively well educated, living in the western areas of the country China, or with nucleus/extensive families were more likely to have early sex in their puberties. Conclusion Sexual education should be strengthened especially in the western areas, and targeted on the students whose parents with relatively higher educational background or who were from extensive families.

OBJECTIVETo explore the therapeutic effectiveness of Ilizarov technique in treatment of infected tibial defects combined with overlaying skin defects.

METHODSTwenty-one cases with infected tibial defects combined with skin defects were treated between 2001 and 2008 includeing 18 males and 3 females with an average age of 31 years ranging from 19 to 43 years. The length of bone defect ranged from 3 to 13 cm (means 6 cm). Skin defect area was from 3 cm x 3 cm to 6 cm x 10 cm; 11 cases combined with drop foot, 5 cases with arthrocleisis of knee. Preoperative X-ray of the affected limb was performed and zone of skin necrosis was marked, then the point and length of osteotomized bone, and scope of bone and soft tissue need for removing were determined. The internal fixation were removed. Opening irrigation, vacuum sealing drainage (VSD), and dressing changing were appllied. The skin was fixed with Kirschner wire and bone was transferred with Ilizarov technique in all patients. The lengthening of bone and skin was carried out for 4 to 7 days after surgery, 1/6 to 1/4 mm once, 4 to 6 times a day. The clinical effectiveness was determined mainly through wound and lengthening of skin.

RESULTSAll patients were followed up for from 6 to 62 months (means 49.5 months). Fourteen of 21 cases received one stage treatment, there was still secretion from end of bone in 3 patients whose bone healed after debridement, the other 4 patients were cured via trimming end of bone and compression fusion. The defects of bone were extended to full length in 18 patients. Abutting end was slightly absorbed and became rattailed in 2 cases, there was lack of blood supply to abutting ends in one patient who was cured via bone graft from iliac bone. Skin defects was cured in 18 patients with one stage treatment, the other 3 patients were cured after infection was controlled. The deformity of drop foot were corrected in 11 patients, and function of knee was improved in five patients. The external fixator was removed at 1.2 to 2.6 years after surgery. At last, bone infections were cured, defects of bone and skin recovered in all patients.

CONCLUSIONOne stage treatment of infected tibial defects combined with skin defects using Ilizarov technique has minimal invasion with less complex surgeries, could reduce the time and expense of treatment.

Adult ; Female ; Fractures, Open ; surgery ; Humans ; Ilizarov Technique ; Male ; Necrosis ; Postoperative Complications ; surgery ; Skin ; pathology ; Soft Tissue Injuries ; surgery ; Tibia ; surgery ; Tibial Fractures ; pathology ; surgery

OBJECTIVETo observe the effect of pulmonary arterial perfusion with Shenqi Fuzheng Injection (SFI)on lung injury during cardiopulmonary bypass (CPB).

METHODSTwenty-two patients with cardiac valvular disease and pulmonary hypertension were randomly divided into the control and the SFI group equally. SFI mixed pure oxygenated blood to the SFI group, and oxygenated blood alone to the control group was perfused via pulmonary artery during CPB. Plasma malondialdehyde (MDA), pulmonary vascular resistance (PVR), ratio of leucocyte counts in venous and arterial blood, and time of mechanical ventilation applied were measured before and at the end of CPB, and 6th, 24th hours after CPB.

RESULTSAfter treatment, MDA content and PVR were significantly higher than those before CPB (P < 0.05), and reduced to normal level 24 h after CPB in both groups, but the peak levels were lower in the SFI group than those in the control group (P < 0.05). The leucocyte counts ratio in venous and arterial blood were significantly higher at the end of CPB and 6 h later than those before CPB in both groups (P < 0.05), but the increment were lower in the SFI group than those in the control group (P < 0.05). Furthermore, the applying mechanical ventilation time in the SFI group was 16.1 +/- 5.5 h, significantly shorter than that in the control group (29.1 +/- 6.9 h, P < 0.01).

CONCLUSIONPulmonary arterial perfusion with SFI could alleviate the CPB induced lung injury.

Adult ; Aged ; Cardiopulmonary Bypass ; adverse effects ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Heart Valve Diseases ; complications ; surgery ; Humans ; Hypertension, Pulmonary ; etiology ; surgery ; Infusions, Intra-Arterial ; Male ; Malondialdehyde ; blood ; Middle Aged ; Phytotherapy ; Pulmonary Artery ; Respiratory Distress Syndrome, Adult ; blood ; etiology ; prevention & control ; Treatment Outcome ; Vascular Resistance ; drug effects

OBJECTIVETo investigate the effects of the tumor suppressor gene PTC on the growth inhibition and down-regulation of epidermal growth factor receptors (EGFR) in pulmonary adenocarcinoma cell A549.

METHODSPulmonary adenocarcinoma cell A549 were divided into wild type group, mutant type group, blank group and control group. They were transfected with wild-type PTC1 plasmids, mutant-PTC1 plasmids and blank plasmids, respectively. After transfection, the cell growth curve was drown every day for a week. The expression of PTC1 and EGFR were detected by western blot. Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells.

RESULTSAfter transfected with wild-type PTC1, the growth rates of A549 cells were slow down, but the other groups of cells had no change. Compared with the control group, the expression of EGFR were down-regulated. The apoptosis rates in wild type group was 24.5%, and the mutant type group was 8.3% (P < 0.01). But the apoptosis rate of blank group has no change.

CONCLUSIONWild-type PTC1 could induce apoptosis and inhibitory effect on A549 cells.

Adenocarcinoma ; metabolism ; pathology ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Patched Receptors ; Plasmids ; genetics ; Receptor, Epidermal Growth Factor ; metabolism ; Receptors, Cell Surface ; genetics ; metabolism ; Transfection