BACKGROUND: Mesenchymal stem cells (MSCs), with low immunogenicity, can regulate cellular immunity and mitigate graft rejection, which has a good prospect in tissue engineering. However, it is rarely present in bone marrow. OBJECTIVE: To explore an isolation and culture method of the rabbit bone marrow-derived MSCs, to observe the biological characteristics and differentiation potential of bone marrow-derived MSCs.METHODS: MSCs were isolated from rabbit tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferation in vitro. Morphology was examined by phase contrast microscopy, and the growth curve of cultured MSCs was drawn via MTT results. MSCs were treated with osteogenetic inductor (L-DMEM/F12, 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 200 μmol/L vitamin C, 10 mmol/L β-phosphoglycerol), adipose inductor (L-DMEM/F12, 10% FBS, 1 μmol/L dexamethasone, 200 μmol/L antifani, 0.5 mmol/L IBMX, 10 μg/mL insulin), and chondrocytes inductor (L-DMEM/F12, 10% FBS, 10 μg/L TGF-β1, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C, 6.25 mg/L insulin) to differentiated into osteoblast, dipocytes and chondrocytes. And the differentiated cells were identified by alkaline phosphatase staining, oil red O staining, and toluidine blue staining, respectively.RESULTS AND CONCLUSION: Bone marrow-derived MSCs can be isolated and cultured by the combination of gradient centrifugation and different adherent method in vitro, which have the better potentiality of proliferation and multi-directional differentiation. Mostly of the primary and passaged cells were spindle-shaped. After osteogenetic induction, cells were positive to alkaline phosphatase staining. Oil red O staining showed that red lipid droplet existed in adipose cells, and toluidine blue staining showed that toluidine blue was positive after chondrocytes induction.

OBJECTIVETo investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the association between cell proliferation and apoptosis and the metabolic status.

METHODSAn in vitro hypoxic cell model was established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and apoptosis of the cells were examined using MTT assay and flow cytometry, respectively.

RESULTSThe cells in hypoxic culture showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential.

CONCLUSIONHypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell death is directly correlated with their metabolic status.

Adenosine Triphosphate ; metabolism ; Apoptosis ; Carrier Proteins ; metabolism ; Cell Hypoxia ; Cell Line, Tumor ; Cell Proliferation ; Central Nervous System Neoplasms ; metabolism ; pathology ; Glioma ; metabolism ; pathology ; Glycolysis ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Membrane Potential, Mitochondrial ; Membrane Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Thyroid Hormones ; metabolism

Objective The aim of this study was to investigate the expression of cell cycle checkpoint kinase 1(Chk1)gene in glioblastoma cells( GBM) and its correlation with GBM cell proliferation,tumorigenic activity and prognosis. Methods The ex-pression of Chk1 in GBM cells was selected and analyzed by TCGA database and brain tumor molecular database( Rembrandt),and the level of Chk1 expression in GBM cells was detected by molecular biology techniques such as Western blot and Real-Time PCR. The expression of Chk1 was silenced by siRNA to investigate its effect on proliferation and colony-forming ability of GBM cells. The prognosis survival of GBM patients accompanying with Chk1 expression was analyzed by immunohistochemical staining and Rembrandt database. Results The results of TCGA database and Rembrandt showed that Chk1 gene was highly expressed in GBM tissues. West-ern blot and Real-Time PCR also showed that Chk1 gene was highly expressed in GBM cells. Lentiviral transfection siRNA-specific silencing of Chk1 significantly inhibited proliferation and colony-forming ability of U87 cells( P<0. 01 and P<0. 05). Prognostic survival analysis showed that GBM patients with low expression of Chk1 gene had a significantly better clinical outcome than those of GBM patients with high expression of Chk1 gene(P<0. 001). Conclusion Chk1 gene is overexpressed in GBM cells,up-regula-tion of Chk1 gene expression can promote the growth and proliferation of GBM cells,and Chk1 gene is associated with poor prognosis in GBM patients.

OBJECTIVE：To preliminarily investigate the improveme nt effects and mechanism of Mongolian medicine Saorilao-4 decoction on specific pulmonary fibrosis model rats. METHODS ：Male SD rats were randomly divided into normal control group ，model group ，positive control group （pirfenidone，0.163 g/kg）and Saorilao- 4 decoction low ，medium and high dose groups （0.899，1.798，3.596 g/kg），8 rats in each group. Except for normal control group ，other groups were given 6 mg/mL bleomycin intratracheally at 5 mg/kg once to induce the specific pulmonary fibrosis model. From the first day after modeling ， normal control group and model group were given normal saline intragastrically ，other groups were given corresponding drugs intragastrically，once a day ，10 mL/kg，for 4 weeks. During the experimental period ，the general condition of the rats in each group was observed and the body mass was weighed. Twenty-four h after last medication ，the appearance morphology of rat l ung in each group were observed. The morphological characteristics of lung tissues were observed by HE and Masson staining. ELISA was adopted to determine the activity of SOD and the content of MDA in serum ，the contents of hydroxyproline （HYP），IL-1β，IL-6，hyaluronidase（HA），laminin（LN）precollagen type Ⅲ（PC-Ⅲ）and collagen type Ⅳ（Col-Ⅳ）in lung tissue. RT-PCR was used to determine the mRNA 发。E-mail：bwf007007@sina.com expression of TGF-β 1，Smad3 and Smad 7 in lung tissue. RESULTS：Compared with model group ，the activity ，hair and diet of the rats in each dose group of Saorilao- 4 decoction and positive control group were significant ly improved ，and the body mass after the last administration was significantly increased ； the pathological change of lung and pulmonary fibrosis were significantly improved ，and the activity of SOD in serum was increased significantly. Serum content of MDA （except for Saorilao- 4 decoction medium dose group ），the contents of HYP （except for Saorilao- 4 decoction high dose group ），IL-1β，IL-6，HA，LN，PC-Ⅲ，Col-Ⅳ（except for Saorilao- 4 decoction high dose group）as well as mRNA expression of TGF-β1 and Smad 3 in lung tissue were significantly decreased ；mRNA expression of Smad 7 was significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Saorilao-4 decoction can significantly improve the lung pathological changes ，delay and reverse the progression of pulmonary fibrosis in specific pulmonary fibrosis model rats ，the mechanism of which may be related to the inhibition of inflammatory response ， improvement of lipid peroxidation ， down-regulation of TGF-β1 and Smad 3 mRNA expression ，and up-regulation of Smad 7 mRNA expression.

【Objective】 To explore the efficacy of Mathieu combined with tongue-shaped flap covering in children with distal hypospadias and micropenis head deformity. 【Methods】 The clinical data of 72 patients with distal hypospadias complicated with micropenis treated during Jan.2018 and Jun.2021 were retrospectively analyzed, including 35 patients who underwent modified Mathieu combined with tongue-shaped flap and external urethral orificium (study group), and 37 patients who underwent traditional Mathieu (control group). Urethral stricture, urethral fistula, urethral diverticulum and penile head cleft were compared between the two groups. 【Results】 Urinary fistula occurred in 2 cases (5.27%) and 9 cases (24.32%) in the study group and control group, respectively, the incidence being much lower in the study group (P=0.028). Urethral stricture occurred in 1 case (2.86%) and 1 case (2.70%) in the study group and control group, respectively; penile head dehiscence in 1 case (2.86%) and 3 cases (8.11%); urethral diverticulum in 0 case (0%) and 2 cases (5.41%). There were no significant differences between the two groups in the incidence of urethral stricture, urethral dehiscence and urethral diverticulum (P>0.05). 【Conclusion】 Mathieu combined with tongue-shaped flap covering to treat children with distal hypospadias with micropenis head deformity can reduce the incidence of postoperative complications, achieve high surgical satisfaction and appearance satisfaction in the first phase, reduce harm caused by surgery, and promote patients’ psychological health.