OBJECTIVETo investigate the role of RLIP76 in regulating multi-drug resistance in small cell lung cancer (SCLC), and to analyze the relationship between its expression and prognosis.

METHODSThe expressions of RLIP76 protein and gene were detected by Western blotting and real-time PCR (RT-PCR) in both the chemosensitive SCLC H69 cell line and chemoresistant H69AR cell line, respectively. siRNA was transfected into the H69AR cells to inhibit RLIP76 expression, and eGFP-RLIP76 was transfected into the H69 cells to enhance RLIP76 expression. The drug-sensitivity of cells to chemotherapeutic drugs (ADM, DDP, VP-16) were detected by CCK8 assay. The expression of RLIP76 in the SCLC tissues was detected by immunohistochemistry. The relationship of RLIP76 expression with clinicopathological features and prognosis of the patients was analyzed.

RESULTSThe expression of RLIP76 in H69AR cells was 13.675 ± 0.983, significantly higher than 1.074 ± 0.107 in the H69 cells (P < 0.01). The drug-sensitivities of H69AR cells to chemotherapeutic drugs were significantly increased when the expression of RLIP76 was down-regulated (P< 0.001). The sensitivities of H69 cells to chemotherapeutic drugs ADM, DDP and VP-16 were significantly decreased after transfection with eGFP-RLIP76 up-regulating the RLIP76 expression (P = 0.003). The positive expression rates were 61.3% and 9.4% in the SCLC tumor tissues and para-cancerous tissues, respectively (P < 0.01). The expression of RLIP76 was significantly correlated with clinical stage, chemosensitivity and overall survival of the SCLC patients (P < 0.05).

CONCLUSIONSOur results suggest that RLIP76 is involved in the regulation of small cell lung cancer multidrug resistance. RLIP76 may serve as a potential target gene to evaluate the chemosensitivity and clinical prognostic for small cell lung cancer.

ATP-Binding Cassette Transporters ; metabolism ; physiology ; Antineoplastic Agents ; pharmacology ; Cisplatin ; pharmacology ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Etoposide ; pharmacology ; GTPase-Activating Proteins ; metabolism ; physiology ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Small Cell Lung Carcinoma ; drug therapy ; metabolism ; Transfection ; Up-Regulation

Objective To investigate the clinical significance of YMDD mutation during Lamivudine therapy on chronic hepatitis B.Methods Fluorometric analysis PCR, ELISA were used to estimate the YMDD mutation, HBVDNA quantative level and HBeAg for HBV of 72 cases with chronic hepatitis B before therapy (0 month), and after Lamivudine therapy for 9,12,18 months.Results The YMDD mutation was not observed in these cases before Lamivudine therapy. The mutation was found in 8 cases (11.1%), 17 cases (23.6%) and 28 cases (38.9%) at 9, 12, 18 month for therapy. The YMDD mutation rate rose with the therapy time lasting (P＜0.05). Moreover, the YmDD mutation rate in the patients with HBVDNA quantity higher than 108 copies/ml was significantly higher than that in the patients with HBVDNA quantity lower than 108 copies/ml (P＜0.005). The YMDD variation rate in patients with HBeAg positive and in patients with HBeAg negative showed no significant difference (P＞0.05). The HBeAg negative conversion rate was significantly higher in non-mutation group than that in mutation group (P＜0.05).Conclusion The serum virus quantity may be regard as an early estimate indication of the development of YMDD mutation during Lamivudine therapy.

Objective To explore the relationships of serum levels of leptin,TNF-?,FFA and resistin with IR and between themselves. Methods 48 T2DM patients and 47 non-diabetic controls were selected.Levels of leptin,TNF-?,FFA and resistin were measured.FPG,insulin,blood lipid,blood pressure,BMI and WHR were measured.Results The levels of leptin,TNF-?,FFA and resistin were correlated positively with HOMA-IR(P

Objective To investigate the effects and significance of neuronavigation and electrocorticography monitoring in resection of eloquent brain glioma. Methods Thirty-six cases with intracranial tumors accepted microneurosurgery resection under neuronavigation and electrocorticography monitoring. The clinical data and postoperative outcome were analyzed. Results The mean registration error was (2.0 ±0. 5)mm in all operations and all skin flaps and bone windows designed by neuronavigation could fit the operation demands. Total resectin of the tumor was achieved in 31 cases and subtotal resection in 5 cases. Neurological symptoms improved and no severe complications or death happened in all patients. Conclusion Neuronavigation combined with electrocorticography monitoring can accurately locate the eloquent glioma and retrieve the brain shift. This method is a real-time technique and has functional test ability. It can improve the total removal rate and decrease the mortality and disabled rate.

Objective To improve the preoperative diagnosis accuracy of cerebellopontine angle tumors through analyzing MRI findings. Methods Ninety-six cases with cerebellopontine angle tumors, confirmed by pathology and surgery, were collected and underwent MRI scan plus enhanced MRI. Among the 96 capes, we observed acoustic neurinoma in 55 cases, meningioma in 20 cases, cholesteatoma in 9 cases, trigeminal neuroma in 7 cases,cavernous hemangioma in 3 cases,arachnoid cyst in 2 cases. The MRI characteristics of all cases were analyzed retrospectively. Results The chief type of tumor happened in the cerebellopontine angle zone was acoustic neurinoma,followed in order by meningioma,cholesteatoma,trigeminal neuroma,cavernous hemangioma and arachnoid cyst. The accuracy of preoperative localization and qualitative diagnosis were 100% and 94.7%respectively.Conclusion MRI has a high value in the diagnosis and differential diagnosis of cerebellopontine angle tumors,which can be used as a preferred preoperative examination method in cerebellopontine angle tumors.

Objective To investigate the potential role of MCP-1/CCL2 in experimental acute necrotizing pancreatitis (ANP) and complications. Methods 60 SD male rats were randomly divided into 3 groups: sham operation group ( n = 20 ), ANP group ( n = 20 ) and MCP-1 group ( n = 20 ). ANP model was induced by retrograde infusion of 3.5% sodium taurocholate, MCP-1 group received subcutaneous injection of MCP-1 antibody 0 h and 6 h after ANP induction. The serum levels of amylase, MCP-1, D-lactic acid,histological changes and the expression of MCP-1 mRNA of lung, small intestine and pancreas, the expression of MCP-1 protein in pancreas, MPO levels of small intestine MPO were determined. Results The serum levels of amylase, MCP-1, D-lactic acid in MCP-1 group at 12 h were (4666 ±412)U/L, (39.53 ±8.25)pg/ml and (6.3 ±2.2)mg/L, which were significantly lower than those in ANP group [ (9611 ±363)U/L, (63.42 ±9.32) pg/ml, (9.3 ± 2. 1 ) mg/L, P< 0.05 ) ]; the expression of MCP-1 mRNA in pancreas, small intestine and lung were 0.431 ± 0.009, 0. 211 ± 0.018 and 0.442 ± 0.017, which were significantly lower than those in ANP group [ (0.624 ±0. 010, 0. 523 ±0. 019 and 0. 569 ±0. 024, P <0.05) ]; the expression of MCP-1 protein in pancreas was 2.0 ± 0. 1, which was significantly lower than that in ANP group (4. 0 ± 0. 2, P <0.05). Lung and small intestine MPO were (11.1 ±3.0)U/g and ( 19.2 ±2.0)U/g, which were significantly lower than those in ANP group[(39.2±3.1)U/g and(13.1±2.1)U/g, P<0.05]. Conclusions Early blockade of MCP-1 not only attenuates the severity of ANP, but also decreases the degree of acute lung injury and intestine barrier dysfunction.

BACKGROUND:Choosing an effective means to label and trace the distribution, differentiation and migration of celsin vivo help to further explore the specific mechanism of cels that exert a therapeutic effect. OBJECTIVE:To understand the migration and localization of BrdU-labeled human umbilical cord mesenchymal stem cels in brain injury model rats. METHODS:Human umbilical cord blood samples were obtained, and the isolation of human umbilical cord mesenchymal stem cels was carried out. The primary and passage culture were performed. The phenotype of cels was detected by flow cytometry. Passage 3 human umbilical cord mesenchymal stem cels were labeled using BrdU, and the cel proliferation was detected using MTT method. BrdU-labeled cels were injected into brain injury ratsvia the tail vein. At 14 days after transplantation, brain tissues in the injury region were cut into sections and the migration and location of the umbilical cord mesenchymal stem cels were observed under inverted
fluorescence microscope. RESULTS AND CONCLUSION: Cel surface specific markers CD45 and CD34 were detected by flow cytometry, but the cels could not express CD44, CD105 and CD29. Based on the cel growth curve, the cels came into a conditioning period at 1-3 days of seeding and came into a logarithmic phase at 3-5 days. BrdU-positive cels were visible at the injury region after 14 days, indicating that in the rats, transplanted human umbilical cord mesenchymal stem cels migrated from the peripheral blood to the site of brain injury to achieve the effective repair of injured parts. Cite this article:Liu HL, Liu ZJ, Chen XB, Hu WZ, Ding BQ. Migration and localization of umbilical cord mesenchymal stem cels implanted into brain injury model rats. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):31-35.

BACKGROUND:Bone marrow stromal cel s can differentiate into nerve cel s to promote nerve tissue repair, but the exact mechanism has not been ful y elucidated.
OBJECTIVE:To explore the influence of adenovirus-mediatedβnerve growth factor transfection on bone marrow stromal stem cel transplantation fighting against brain injury in rats.
METHODS:(1) Rat bone marrow stromal stem cel s were cultured in vitro, transfected with the adenovirus-mediatedβnerve growth factor and directional y induced usingβ-mercaptoethanol. (2) A total of 210 Sprague-Dawley rats were randomized into induction+tranfection group, induction+non-transfection group, induction+medium group, model group, and sham group (n=42 per group). Rat skul injury models were made, and given corresponding treatments at different time points (12, 24, 36, 48, 72 hours). Neurological function of rats was evaluated based on neurological severity scores on the day that the rats were given transplantation, and 1, 2, 3, 4 weeks after transplantation. (3) Another 75 Sprague-Dawley rats were also divided into five groups (n=15 per group) as above, fol owed by model establishment and corresponding treatments at 24 hours after modeling. Neurological severity scores were recorded at the same day, 1, 2, 3, 4 weeks after transplantation. Five rats from each group were sacrificed to detect levels of malondialdehyde and superoxide dismutase in the rat brain at the same day, 2 and 4 weeks after transplantation, respectively.
RESULTS AND CONCLUSION:If the cel s were transplanted within 48 hours after modeling, the neurological severity scores in the induction+transfection group decreased significantly compared with the induction+non-transfection group and model group at 1 and 2 weeks after transplantation (P<0.05). If the cel s were transplanted at different time, the neurological severity scores in the induction+transfection group were decreased significantly compared with the induction+non-transfection group and model group at 3 and 4 weeks after transplantation (P<0.05). If the cel s were transplanted within 24 hours after modeling, the neurological severity scores in the induction+transfection group decreased significantly compared with the model group at 1 week after transplantation (P<0.05), and the neurological severity scores in the induction+transfection group and induction+non-transfection group both were significantly lower than those in the model group (P<0.05). Two weeks after cel transplantation, the level of superoxide dismutase was significantly higher in the induction+transfection group than the induction+medium group and model group (P<0.05), but the level of malondialdehyde was significantly lower (P<0.05). Al these findings indicate that adenovirus-mediatedβnerve growth factor transfer plays a certain neuroprotective role in bone marrow stromal stem cel transplantation for brain injury in rats.

BACKGROUND:Stem celltherapy for renal ischemia-reperfusion injury has been the hot topics for many scholars. Its mechanism is very complex, which could not be explained by simple mechanism of stem cells differentiation. It is the result involving a variety of mechanisms. OBJECTIVE:To investigate the influence on immune cells during the bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury, then to preliminary summarize the immune regulation mechanism of bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury. METHODS:First, we established a model of renal ischemia-reperfusion injury in rats and, cultured and purified rat bone marrow mesenchymal stem cells in vitro. Then, the bone marrow mesenchymal stem cells were transplanted into the rat models. Using flow cytometry detection technology, we analyzed the proportion of CD4+CD25+regulatory T cells of rat spleen cells, discussed the effects on immune cells during the bone marrow mesenchymal stem celltherapy for renal ischemia-reperfusion injury, and then transferred the rat’s spleen cells to the nude mice which were subjected to renal renal ischemia-reperfusion injury. Renal function and renal histological changes of nude mice were assessed. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem celltransplantation could significantly inhibit the decrease of CD4+CD25+regulatory T cellof spleen cells in rats with renal ischemia-reperfusion injury. The transplantation of spleen cells from the above-mentioned rats to nude mice could obviously protect nude mice from renal ischemia-reperfusion injury, characterized by lower serum creatinine, blood urea nitrogen and renal tubule pathologic damage score. Therefore, bone marrow mesenchymal stem cells have protective effects on renal ischemia-reperfusion injury by regulating the immune system.

Objective:To investigate the possible role of miR-181d in regulating the multidrug resistance (MDR) of small cell lung cancer (SCLC) and its clinical significance. Methods: Quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) and Western blot were used to investigate the differential expression of miR-181d and BCL2 from mRNA and protein levels in the chemo-sensitivity cell H69 and the chemo-resistance cell H69AR. The miR-181d expression in H69AR was then upregulated. More-over, CCK8 assay was employed to detect the sensitivities of the cells to chemotherapy drugs, such as ADM, DDP, and VP-16. Mean-while, the expression of miR-181d in the specimens of 87 cases with SCLC were detected using QRT-PCR. All patients received the chemotherapeutic regimen of EP (etoposide+cisplatin). Correlation of the miR-181d expression with clinicopathological features, prog-nosis, and survival time of the patients was studied. Results:miR-181d was downregulated in the SCLC multidrug-resistant cell line H69AR and chemo-resistant patients. Moreover, miR-181d was concurrent with the upregulation of BCL2 protein compared with the parental H69 cell line and chemo-sensitive patients (P<0.001). miR-181d expression in H69 cells resistant to chemotherapy drugs (ADM, DDP, and VP-16) was inhibited (P<0.01). Enforced miR-181d expression reduced the BCL2 protein level and sensitized H69AR cells to chemotherapy drugs (P<0.01). miR-181d expression was associated with tumor stage, sensitivity of chemotherapy, and survival time (all P<0.001). Patients with high miR-181d expression had longer overall survival and progress-free survival time com-pared with those with low miR-181d expression (P<0.001). Conclusion: miR-181d may play a role in the development of MDR in SCLC and may be a potential predictive factor for treatment efficacy.