Objective To study the clinical efficacy of emergency early mechanical ventilation and bron -choalveolar lavage and drug intervention to severe asthma .Methods 140 cases with severe asthma were selected ,the patients were divided into the study group and the control group according to a random number table ,70 cases in each group,the control group were received early mechanical ventilation ,the study group were given bronchoalveolar lavage and interventional therapy on the basis of the control group ,clinical efficacy ,alveolar function ,her condition improved and hospital stay of the two groups were compared .Results The total efficiency of the study group were 92.9%(65/70) significantly higher than 81.4%(57/70) in the control group,the difference was statistically significant (χ2 =12.382,P<0.05);Forced expiratory volume in one second (FEV1),forced vital capacity (FVC) and FEV1/FVC of the study group were significantly better than that of the the control group ,the difference was statistically significant ( t=9.098,10.382,11.204,all P<0.05);Condition improved and hospital stay of the study group were significantly shorter than that of the control group,the difference was statistically significant (t=9.898,12.393,all P<0.05). Conclusion Emergency early mechanical ventilation and bronchoalveolar lavage and drug intervention to severe asthma had better clinical efficacy ,which can significantly improve the clinical symptoms of the patient .

Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

BACKGROUND:Rapid development of spermatogonial stem cells is the new hope for assisting reproductive technologies,and the stability of the number and structure of the chromosome cultured in vitro is one of the important factors in usage.OBJECTIVE: To study the influential factors on G-band exhibition in mouse spermatogonial stem cells, so as to offer a technology to identify karyotype of stem cells in culture.DESIGN: Observational experiment.SETTING: Luzhou Medical College.MATERIALS: The experiment was conducted in the Laboratory of Medical Molecular Biology, Luzhou Medical College from March 2004 to April 2005. 10 days old male and female Kunming mice were provided by Department of Animal of Luzhou Medical College (number of license No. 17 of experimental animal quality administration in Sichuan province).There were low-sugar DMEM medium, 10 mg/L Mitomycin-C, IMDM medium, 1 ×10-5 mol/L colchicines (PBS allocation),7.5 mmol/L KCL, fixative (mixture glacial acetic acid with methanol in 1:3), Giemsa staining solution and 0.25% zymine.METHODS: Bone marrow was aspirated from the thigh bone of mouse for feeder layer cells preparation. Cells from the male mice testis of 7-8 days after birth were prepared and made into cell suspension. After adjusting the cell density to 3×105 L-1, they were inoculated into the feeder layer of bone marrow stromal cells. Cells were cultured at 37 ℃ in CO2 incubator containing CO2 of 0.05 volume fraction and 70% humidity. The proliferation groups of stem cells cultured 15-20 days were selected, stirred and spread, and then treated with colchicine for 4-6 hours. Cell suspension was collected,and then stained after hypotonic treatment. The cells whose chromosomes were dispersed well and in metaphase were selected, and number of chromosomes was counted, and then the morphology of chromosomes was observed.MAIN OUTCOME MEASURES: Culture of bone marrow stromal cells and spermatogonial stem cells and coloration and count of chromosomes.RESULTS: The karyotype of spermatogonial stem cells was the same as the body cells of normal mouse, which showed granule or rod-shape. Karyotype was 20 pairs, 40 bars. Three kinds of chromosome morphology could be observed under oil immersion lens. The first type was condensed, which could be counted in total, but the band could not be seen.The second type was chromatosome that spread completely in the center of equatorial plate and were in metaphase. In this phase, total numbers and band were seen clearly. Last type-chromosomes had already folded and moved towards two poles and concentrated gradually, the total number of chromosomes could be counted, but bending could not be seen clearly.CONCLUSION:Many factors can affect the karyotype of spermatogonial stem cells, including the phase of cell division,effect of hypotonic solution, diffusion of the cell when dropping slides, concentration of trypsin and digestion time, etc.

Objective Off-pump coronary artery bypass grafting (OPCAB) is used more widely in recent years in China. However, there is an argument on benefits and risks of off-pump surgery. Many studies shown that OPCAB had more benefits in short-term outcomes than conventional coronary artery bypass grafting(CCABG). But evidences from other studies suggested that OPCAB resulted in less long-term graft patency as compared with on-pump surgery. This study examined the longterm graft patency of OPCAB and CCABG performed by one surgeon. Methods 50 patients who had received surgical revascularization by a surgeon for more than 5 years were reviewed, 25 patients received conventional coronary artery bypass grafting ( group 1 ) and 25 patients received OPCAB ( group 2). All patients had angiograms for compareing the graft patency between the two groups. Results Among 25 patients in group 1,21 were male and 4 were female. The mean age of patients at surgery was (55.4 ±8.9) years. 15 cases had unstable angina, 16 patients had old myocardial infarction and 6 cases had diabetes.The ejection fraction (EF) was 0.58 ±0.14. The mean number of bypasses per patient was 3.32 ±0.63. Mean duration of operation was (3.58 ± 0. 82) hours. Mean follow-up duration was ( 86.52 ± 12.48) months. 83 grafts were evaluated for patency ( open vs. closed) and were graded by Fitzgibbon as grade A ( excellent graft), B ( impaired graft, with a stenosis of ≥50%, or a diameter less than 50% of the grafted artery), or O ( completely occluded). The graft patency was 73.49%, 61grafts were graded as Fitzgibbon A, 6 grafts as Fitzgibbon B and 16 grafts as Fitzgibbon 0. 25 patients were in group 2, 21males and 4 females. The mean age of patients at procedure was (58.2 ± 9.09) years, 11 patients had unstable angina, 13 patients had old myocardial infarction and 6 cases had diabetes. The ejection fraction (EF) was 0.59 ± 0. 14. Conclusion No 2011.03.013 difference in long-term graft patency was identified between on-pump and off-pump coronary artery bypass grafting. Off-pump oronary artery bypass grafting preformed by an experienced surgeon may gain similar long-term graft patency to that of conventional bypass.

OBJECTIVETo investigate the application of relay flaps pedicled by perforator from digital artery for reconstruction of soft tissue defects at finger tip.

METHODSFrom Mar. 2012 to Jun. 2014, 9 cases with soft tissue defects at finger tip were reconstructed with relay flaps at one side of finger pedicled by perforator from digital artery. The flap size ranged from 1.3 cm x 1.6 cm to 1.6 cm x 2.2 cm. The defects at donor sites were covered by adjacent web perforator V-Y advanced flaps.

RESULTSAll the 18 flaps in 9 cases survived completely with primary healing both in recipient and donor sites. The patients were followed up for 5 months to 2 years ( average, 12 months) with good elasticity and cosmetic results. No pain happened in the treated finger. The 2-point discrimination distance was 7-8 mm in fingertip flaps, and 10-12 mm in web perforator flaps. Hand function was graded as excellent in 7 cases, good in 2 cases, based on ATM assessment. The affected fingers had normal temperature and cold-resistance during winter. The width and depth of web in the donor site were not affected.

CONCLUSIONSThe relay flaps pedicled by perforator from digital artery can be applied for reconstruction of soft tissue defects at finger tip. The procedure is easy with satisfactory results and reservation of main artery. No skin graft is necessary for closure of defects on donor sites.

Arteries ; Elasticity ; Finger Injuries ; surgery ; Fingers ; blood supply ; Follow-Up Studies ; Humans ; Perforator Flap ; transplantation ; Time Factors ; Transplant Donor Site ; Wound Healing

Female ; Humans ; Infant ; Lung ; physiopathology ; Male ; Pneumonia, Viral ; physiopathology ; Respiratory Function Tests ; Respiratory Syncytial Virus Infections ; physiopathology ; Respiratory Syncytial Viruses

Objective To study the clinical application of mediastinal elastic drainage-tube for patients with anastomotic leak after esophagus carcinoma or esophagogastric junction carcinoma surgery.Methods Two hundred and eighty-two cases esophagus carcinoma or esophagogastric junction carcinoma patients were randomly divided into 2 groups.The control group(n=140) were indwelled thoracic drainage tube after operation.However,the experimental group(n=142) were indwelled thoracic drainage tube and mediastinal elastic drainage-tube after operation.The incidence of the esophagogastrostomy fistula,diagnosis time of anastomotic leakage,maximum temperature,time of continuous ferer,total white blood cell count,cardiopulmonary complications,postoperative hospital stay,hospitalization expenses were observed and analyzed.Results (1)The incidence of the esophagogastrostomy fistula in the experimental group and the control group was 9.8%(14/142) and 12.1%(17/140),and there was no significant difference between the two groups(χ2=0.376,P>0.05).The diagnosis time of anastomotic leakage,maximum temperature,time of continuous ferer and total white blood cell count of the experimental group and the control group was (6.4±0.6) d and (10.6±0.6) d,(38.1±0.1)℃ and (39.0±0.2)℃,(72.2±2.8) h and (102.6±3.3) h,(12.6±0.7)×109/L and (19.7±0.6)×109/L after operation,there was significant difference between the two groups(t=2.708,1.662,3.164,1.837,P<0.05).The incidence of pulmonary complication,cordis complication of the experimental group and the control group was 19.7%(28/142) and 32.1%(45/140),18.3%(26/142) and 40.7%(57/140) after operation,there was significant difference between the two groups (χ2=5.077,6.606,P<0.05).The postoperative hospital stay,hospitalization expenses of the experimental group and the control group was (28.1±4.2) d and (45.6±3.9) d,￥(6 8174.7±3206.5) and￥(8 4774.8±4007.3) after operation,there was significant difference between the two groups(t=2.001,1.709,P<0.05).Conclusion The mediastinal elastic drainage-tube for patients with anastomotic leak after esophagus carcinoma or esophagogastric junction carcinoma surgery can not reduce the incidence of the esophagogastrostomy fistula,but which is conducive to the early diagnosis and timely treatment of anastomotic leakage.Meanwhile it can reduce the incidence rate of cardiac and pulmonary complications,shorten the length of stay in hospital,reduce the total cost of hospitalization.

Objective To explore the influence factors of hidden blood loss during the treatment of femoral intertrochanteric fractures with percutaneous compression plate( PCCP) in elder patient. Methods The data of 136 patients with intertrochanteric fracture in our hos-pital from March 2009 to March 2012 treated with percutaneous compression plate were retrospectively analyzed,the effect of age,fracture type,preoperative aspirin,internal diseases on perioperative hidden blood loss, and different haemoglobin ( HGB) levels in patients with dif-ferent distribution were analyzed. Results The mean hidden blood loss volume was 499 mL,which accounted for 92. 54% of the total blood loss. The mean HGB decline was 11 g/L. The rate of on admisson haemoglobin level lower than 80 g/L was 6. 62% in AO type A2. 3 and A3 groups,and the rate of postoperative haemoglobin level lower than 80 g/L was 14. 70%. The average hidden blood loss for AO type A2. 3 (862 mL) and A3 (698 mL) were higher than those for AO types A1 (430 mL),A2. 1 (450 mL) and A2. 2 (415 mL) (P<0. 05). Multi-variate linear regression analysis showed that comminuted fracture and preoperative aspirin treatment were independently associated with in-creased perioperative hidden blood loss. Conclusion It indicated that the overt blood loss was fewer for PCCP operation. Intertrochanteric fracture was the main reason for substantial perioperative hidden blood loss. Types A2. 3 and A3 intertrochanteric fracture were early predic-tive factors of postoperative haemoglobin level lower than 80 g/L in elderly patients.

Objective To explore the expressions of apoptosis gene Bcl-2 (B-cell leukemia-2) and Bad (bcl-xl/bcl-2-associated death promoter) in lung tissue of mice with acute lung injury (ALI). Method Twenty-four BALB/C female mice were randomly divided into control saline group (n = 6) and ALI group (n = 18). The ALI Group was further divided into 3 subgroups with 6 mice in each subgroup: ALI (4 h) ,ALI (6h) ,and ALI (8h) subgroups. Rats in the normal control group received injection of saline. The ALI models were produced by in-jection of oleic acid (0.9 mL/kg) via vena caudalis, and the criteria were met with the characteristically pathologi-cal changes in the lung tissue. Pathological changes of the lung tissue were examined and scored under light mi-croscopy 4 h,6 h and 8 hours after injury. The expressions of gene Bcl-2 and gene Bad were detected in lung tissue at above set intervals by using RT-PCR. Data of these assays were analyzed by using one-way ANOVA with SPSS version 13.0 software. Statistical significance was established at a P value of less than 0.05. Results The rela-tive magnitude of Bel-2 expression in ALI (4 h), ALI (6 h) and ALI (8 h) subgroups were significantly higher (58.00±5.31), (42.00±4.30), (32.51±10.40) as compared with the control group (24.30±1.00) (F =68.581, P < 0.05). The relative magnitude of Bad expressions in ALI (4 h), ALI(6 h) and ALI (8 h) sub-groups were signiticantly higher (29.32±1.19), (58.64±4.45), (95.12±4.34)as compared with control group (4.01±0.34) (F = 386.902,P < 0.05). The pathological scores ofhmg injury in ALI(4h), ALI (6 h) and ALI (8 h) subgroups were significantly higher (1.82±0.14), (2.52±0.25), (3.45±0.29) as compared with control group (0.27±0.03) (F = 260. 512, P <0.05). Comparisons between groups showed statistical signifi-cances (P < 0.05). Conclusions The aggravation of lung injury induced by oleic acid in mice related to the down-regulation of apoptosis gene Bcl-2 expression and up-regulating apoptosis gene Bad expression in lung tissue.

OBJECTIVE: To study the effects of Tongbi Mixture 2, a compound traditional Chinese herbal medicine for treating rheumatoid arthritis (RA), on immunoregulation of T lymphocytes in rats with collagen-induced arthritis (CIA). METHODS: Forty Wistar rats were randomly divided into normal control group, untreated group, Tongbi Mixture 2-treated group, methotrexate (MTX)-treated group and Tripterygium wilfordii polyglycoside (TWP)-treated group. Except for the rats of the normal control group (injection with normal saline), rats of the other four groups were subcutaneouly multipoint-injected with collagen protein II to induce CIA. The rats were treated with normal saline, Tongbi Mixture 2, MTX tablets and TWP tablets respectively for 36 days. The expressions of CD28 and CD152 were detected by flow cytometry, while the content of tumor necrosis factor-alpha (TNF-alpha) was analyzed by enzyme-linked immunosorbent assay. RESULTS: The expression of CD28 among the three drug treated groups had no statistical difference (P>0.05), while significantly higher than that of the normal control group (P<0.01) and lower than that of the untreated group (P<0.01). The expression of CD152 in the Tongbi Mixture 2 treated-group was lower than those of the MTX- and TWP-treated groups (P<0.05, P<0.01), but had no statistical difference as compared with the normal control group (P>0.05). There were no statistical differences in content of TNF-alpha between the drug treated groups and the normal control group (P>0.05), but the content of TNF-alpha of the drug treated groups was lower than that of the untreated group (P<0.05 or P<0.01). CONCLUSION: Tongbi Mixture 2 can inhibit T lymphocytes through down-regulating the expressions of CD28 and CD152 and the content of TNF-alpha, which may be the major mechanisms in treating RA.