AIM: To investigate the effect of hypoxia on glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ?-actin and endothelin-converting enzyme (ECE)-2 mRNA levels in cultured mouse brain astrocytes (AC). METHODS: AC from neonatal mouse brain was incubated for 24 h in serum-free medium under hypoxic or normoxic conditions. The amount of transferred RNA was estimated using ethidium bromide stained 28S rRNA and 18S rRNA. The levels of tested mRNA were evaluated by Northern blot RNA hybridization. RESULTS: The GAPDH mRNA was up-regulated to 503.0% of the normoxic controls in hypoxic AC (n=10. P

AIM: Humanized-NOD/SCID(hu-NOD/SCID) mouse model was established and the level of immune reconstitution was assessed in this model. METHODS: Mononuclear cells (MNC) and CD34+ cells were isolated or sorted from cord blood(CB). Human CD45, CD19, CD3 markers on cells from NOD/SCID murine peripheral blood(PB), bone marrow(BM), thymus were detected by FCM from 4 to 10 weeks after hematopoietic stem cell transplantation. After 10 weeks, the gene expressions of the human ?2M and RAG2 were detected by RT-PCR in PB or bone marrow of mice model. RESULTS: Human CD45, CD19, CD3 cells populations in PB and BM were found by flow cytometry in mice model transplanted with CD34+ cells or CB MNC from 4 to 10 weeks. The highest positivity of human lymphocytes was at 8 week after transplantation. The levels of human cell engraftment in mice transplanted with CD34+ cells were higher than those in mice transplanted with CB MNC. The mRNA of human ?2M and RAG2 were found by RT-PCR in BM.CONCLUSION: The higher level of human lymphocyte engraftment is established in NOD/SCID mouse model transplanted with CD34+ compared with CB MNC. The maturation of T lymphocytes could be happened in bone marrow of mice model.

Objective To validate genetic suppression of metastastic capability of highly metastastic melanoma cells by endostatin transfection.Method pcDNA3.1-Endo eukaryotic expression vector contained insulin signal peptide sequence was transfected into highly metastatic mice melanoma cell strain B 16.The expression of endostain was detected by RT-PCR and Western blot experiment,melanoma cells were determined with adhere experiment,in vitro invasion and migration experiment and pulmonary metastasis experiment on C57BL/6 mice.Result Endostatin can obviously inhibit the capability of adherence,in vitro invasion and migration and pulmonary metastasis of melanoma cells.Among them,adhere inhibition ratio was 67.3%,in vitro invasion inhibition ratio was 48.4%,cell migration inhibition ratiowas 52.1%and pulmonary metastasis inhibition ratio was 67.3%.Conclusion Endostatin transfection can obviously inhibit the highly metastatie capability of melanoma cells.

Objective To study the effect of short course of mini dose hormonal application after long-segment spinal fusion using allograft bone in scoliosis.Methods A retrospective study was made in 106 patients with scoliosis,who underwent posterior spinal fusion during May,2004 to Jan,2007 in our hospital.These patients were divided into two groups.Hormone group,68 cases,were 13 to 28 years old with a mean age of 16.5 years old.The average fused segments were 9.0 vertebrae(range from 7 to 12 vertebra).Glucocorticoid was used after operation about 5 to7 days.No hormone group,38 eases,were 14 to 30 years old with a mean age of 17.5 years old.The average fused segments also were 9.0 vertebrae(range from 6 to 12 vertebra).No glucocorticoid was used after operation.All patients received spinal instrumentation with TSRH system and spinal fusion with allograft bone.The temperature,white blood cell(WBC),ESR,drainage and complications between two groups after operation were comparative analyzed.Result Between two groups,the variance of the figures of the temperature,WBC,ESR and drainage are statistically significant.There was less complication in hormone group than that in no hormone group.Conclusion After the operation of the segments spinal fusion by allograft bone,using mini-close glucocorticoid by short course of treatment can significantly decrase some complications and ensure spinal fusion.

Objective To observe the different behavior of proliferation and cell cycle of MCF-7 cells when exposured to ropivacaine of different concentrations and further explore its underlying mechanism.Methods Human breast cancer cells MCF-7 were inoculated into culture medium for 24 h,then were randomly divided into four groups:Control group(group C),Ropivacaine 100 μg/ml group(group R1 ),Ropivacaine 200 μg/ml group(group R2),Ropivacaine 400 μg/ml group(group R3).We medicated each group and incubated for 48 h,then detected the cell proliferation and cell cy-cle immediately.The level of protein TCF-4 and beta-catein of groups R3 and C were measured at the same time.Results MCF-7 cell viability of groups R2 and R3 was significantly lowed (P <0.05 ), MCF-7 cell viability of group R1 had no significant difference when compared to group C.G0/G1 phase cells of groups R1,R2 and R3 were significantly less than those of group C,S phase cells of groups R1,R2 and R3 were significantly more than group C,G2/M phase cells of groups R1,R2 and R3 were significantly more than group C (P <0.05).The expression level of TCF-4 and beta-catenin in group R3 was significantly lower than that in group C (P <0.05).Conclusion Ropivacaine inhibits the proliferation of breast cancer cells MCF-7 by down-regulating TCF-4 and beta-cateni.

OBJECTIVEConstruct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.

METHODAPL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.

RESULTThe subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.

CONCLUSIONThe constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.

Apoptosis ; drug effects ; genetics ; Arsenicals ; pharmacology ; Cell Line, Tumor ; DNA, Complementary ; analysis ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; Oxides ; pharmacology ; Sequence Analysis, DNA

Objective To investigate the distribution of TCR Vβ genealogy and clonal expansion in peripheral blood after infusing mesenchymal stem cells (MSC) in patients with chronic GVHD. Methods The complementarity determining region 3 (CDR3) of 24 TCR Vβ subfamily genes in peripheral blood mononuclear cell from 1 case with cGVHD after allogeneic hematopoietic stem cell transplantation (Allo-HSCT),who were treated with infusing MSC,were amplified using RT-PCR. The blood samples were taken at the first and the fifth day after 1st infusion; and the first day,the 10 th day and the 20 th day after the second infusion of MSC,as well as the MSC infused as control . The products were labelled by fluorescein and then analyzed the CDR3 size with gene scan technique to determine the clonality of T cells. Results There were no expression of TCR Vβ subfamily with the MSC infused and after the 1st day of the first infusion of MSC. Then 3,10,14,10 Vβ subfamilies clones are appeared at the other time points,of which were polyclone and oligoclone predominately. In the same time,the manifestations of cCVHD have been abated. Conclusion MSC played a certain role in reviving the immune function of the patients after Allo-HSCT and mitigating the disease of chronic GVHD. Lineage analysis of TCR Vβ subfamily showed some predominant expression.

Objective To explore the effect of oropharyngeal healthy exercise combined with guided education in the treatment of dysphagia in patients with Alzheimer's disease (AD). Methods 90 AD inpatients in the geriatric psychiatric department of Guangzhou Huiai Hospital during May 2014 and August 2015 were equally randomized into the study group and the control group with a random digit table:the former group was treated with routine oral muscles training and the latter with oropharyngeal healthy exercises combined with guided education plus routine muscles training. The comparison was done between the groups in terms of the total curative effectiveness 4 weeks following the intervention. Result The total effective rate in the study group was significantly higher than that in the control group (P<0.01). Conclusion The oropharyngeal healthy exercise combined with guided education can effectively promote the recovery of swallowing function in AD patients.

Objective:To investigate the efficacy of robot-assisted femoral tunnel localization in reconstruction of the medial patellofemoral ligament (MPFL).Methods:A retrospective study was conducted to analyze the 36 patients who had been admitted to Department of Sports Medicine, The Fourth Hospital of Wuhan between January 2019 and January 2022 due to recurrent patellar dislocation. There were 15 males and 21 females; age: 23.5 (18.3, 29.0) years; number of dislocations: 2.5 (2.0, 3.0). They were stratified into 2 cohorts based on utilization of robot-assistance. In the observation group (17 cases), the femoral tunnel localization was robot-assisted in MPFL reconstruction; in the control group (19 cases), the femoral tunnel localization was guided by C-arm fluoroscopy in MPFL reconstruction. The 2 groups were compared in terms of operation time, frequency of guide wire placement, visual analogue scale (VAS) at postoperative 1 d, patellar tilt angle (PTA) and the disparity between actual femoral tunnel insertion and ideal tunnel insertion point (Sch?ttle point) at postoperative 1 to 3 d, and Lysholm knee score and International Knee Documentation Committee (IKDC) score at the last follow-up.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). All patients were followed up for 12.0 (10.3, 13.0) months. In the observation group, the operation time [(64.1±16.7) min], frequency of guide wire placement [1.0 (1.0, 2.0) times], VAS [2.5 (2.0, 3.0) points], and disparity between actual femoral tunnel insertion and ideal tunnel insertion point [(4.7±1.2) mm] were significantly better than those in the control group [(84.2±19.7) min, 3.0 (2.0, 4.0) times, 3.5 (3.0, 4.0) points, and (6.1±1.2) mm] ( P<0.05). There was no statistical difference between the 2 groups in PTA, Lysholm knee score or IKDC score ( P>0.05). Conclusions:The short-term clinical efficacy of robot-assisted femoral tunnel localization is satisfactory in MPFL reconstruction. Compared with the intraoperative C-arm fluoroscopy, robot-assisted localization can decrease the frequency of guide wire placement, enhance femoral tunnel accuracy and efficiency, and alleviate more postoperative pain for the patients.

Objective:To compare the effects of thoracoscopic anatomical segmentectomy and thoracoscopic lobectomy on patients' respiratory function.Methods:Retrospective analysis of 326 patients who underwent thoracoscopic surgery from July 2016 to July 2019(209 patients underwent anatomical segmentectomy, 117 patients underwent lobectomy). According to variables including gender, age, tumor location, smoking history and BMI, two propensity score-matched cohorts including 89 patients respectively were constructed. The patients’ baseline data and respiratory function date of the patients pre-operation and post-operation were analyzed. The measurement data that obey the normal distribution were described by mean±standard deviation, and the t-test was used for comparison between groups; the measurement data of non-normal distribution was described by the median value( P25, P75), and the Wilcoxon rank sum test was used for the comparison between groups; The data was described by frequency, and the chi-square test or Fisher's exact probability method was used for comparison between groups. Results:At the first-month follow-up after surgery, there was no significant difference in the variation of FVC[(0.48±0.40)L vs.(0.34±0.37)L, P=0.215)and FEV1[(0.52±0.46)L vs.(0.43±0.77)L, P=0.364), and in the change rate of FVC(%)[15.23(8.74, 21.25) vs. 14.58(7.75, 19.40), P=0.122], FEV1(%)[17.25(9.56, 22.78) vs. 16.42(9.15, 20.28), P=0.154]and DLCO(%)[18.54(10.88, 25.68)vs. 17.45(9.58, 23.75) P=0.245]. Between the segmentectomy group and lobectomy group, there was a significant difference in the alteration of FVC[(0.50±0.47)L vs. (0.29±0.31)L, P=0.031] and FEV1[(0.44±0.34)L vs.(0.24±0.23)L, P<0.001], the change rate of FVC(%)[14.27(7.87, 22.32) vs. 9.95(5.56, 17.24), P=0.008]、FEV1(%)[15.23(8.36, 22.17)vs. 10.05(5.15, 18.54), P<0.001]and DLCO(%)[13.74(6.24, 19.78) vs. 4.45(-2.32, 13.75), P=0.023]in the 6th month after surgery. The lobectomy group had a higher variation of FEV1[(0.34±0.49)L vs.(0.18±0.26)L, P=0.006] and change rate of FVC(%)[9.28(2.15, 18.94) vs. 5.24(0.52, 11.45), P=0.0032] and FEV1(%)[10.45(3.15, 21.32) vs. 6.50(1.55, 14.24), P<0.001] in the first year after surgery. However, the variation of FVC[(0.29±0.36)L vs.(0.21±0.24)L, P=0.176) and the change rate of DLCO(%)[8.35(2.15, 16.45) vs. 6.23(2.12, 14.54), P=0.143] didn't show a significant difference between the two groups. Conclusion:Whether in the short or the middle postoperative period, segmentectomy can preserve postoperative respiratory function than lobectomy.