OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

OBJECTIVETo observe the therapeutic effect of Fu'an Liquid (FAL) for retention enema in treating gastrointestinal (GI) dysfunction of children with critical illness.

METHODSEighty-nine patients were randomly divided into two groups, 52 in the treated group and 37 in the control group. Conventional therapy of western medicine was given to both groups and to the treated group FAL was given additionally. Plasma endothelin (ET) level was measured during admission, GI dysfunction occurrence and after treatment, and the therapeutic effect as well as the recovery of GI condition were observed.

RESULTSThe total effective rate of FAL in treating GI dysfunction was 84.62%, which was significantly higher than that in the control group (62.17%) (P < 0.05). In the treated group, 34 cases were treated successfully, 16 died and the other 2 abandoned, the mortality rate being 30.77%, while in the control group, the corresponding numbers were 16, 18, 3 and 48.65%. The mortality rate in the treated group was lower than that in the control group (chi 2 = 4.64, P < 0.05). Level of ET in both groups was higher than normal range during admission (P < 0.01), it further increased when GI dysfunction occurred (P < 0.01), and decreased when successfully treated, the decrease was quicker in the treated group than that in the control group (P < 0.05).

CONCLUSIONIn children with critical illness, ET level would increase when the patient was complicated with GI dysfunction. FAL for retention enema could reduce the ET level effectively, promote the recovery of patients from GI dysfunction, so as to play a definite role in enhancing the successful rate of rescue.

Administration, Rectal ; Adolescent ; Child ; Child, Preschool ; Critical Care ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gastrointestinal Diseases ; drug therapy ; etiology ; Humans ; Infant ; Infant, Newborn ; Male ; Multiple Organ Failure ; complications ; drug therapy ; Phytotherapy ; Pneumonia ; complications ; Rheum ; Shock, Septic ; complications

Objective To identify the important risk factors for type 2 Diabetes Mellitus (T2DM) and develop effective strategies to address the problem of T2DM.Our study aimed to evaluate the association between apolipoprotein E (ApoE) genetic polymorphism and type 2 diabetes,and to provide clues for the etiology of T2DM.Methods Based on the criteria of inclusion and exclusion,we extracted,pooled,analyzed and assessed the case-control studies of ApoE polymorphism and T2DM published in PubMed,Web of Science,Medline,WanFang,VIP,and CNKI databases by R soft-ware (version 3.4.3).We used Random-effect models when heterogeneity was present in between-study,and fixed-effect models otherwise.Results We had 59 studies covering 6,872 cases with T2DM and 8,250 controls,and compared the alleles and genotypes of ApoE between cases and controls.When we conducted a comparison between ApoE ε4 and ε3 alleles,we produced a pooled OR of 1.18 (95％ CI:1.09-1.28;P ＜ 0.001).ApoE ε2/ε2 genotype displayed a possible association with T2DM (OR =1.46;95％ CI:1.11-1.93;P =0.007),ε3/ε4 genotype showed a 1.11-fold risk (OR =1.11;95％ CI:1.01-1.22;P =0.039) and ε4/ε4 genotype had a 1.71-fold risk of developing T2DM (OR =1.71;95％ CI:1.33-2.19;P ＜ 0.001) when they were compared with ε3/ε3 genotype.Conclusions There is an association between ApoE polymorphism and T2DM:allele ε4 and genotypes (ε2/ε2,ε3/ε4,and ε4/ε4) are associated with the increased risk for the development of T2DM,and they may be risk factors for T2DM.

OBJECTIVETo investigate the different expressions of cytoskeletal organizer ezrin and cytoskeleton protein beta- and gamma-actin in hepatocellular carcinoma (HCC) cell lines with different metastatic potentials and to explore the role of ezrin in cell growth and metastasis in HCC cell lines SF7721 and MHCC97-H.

METHODSImmunofluorescence, RT-PCR and Western blot were used to detect the gene and protein expressions of ezrin and actin in hepatocellular carcinoma cell lines with different metastatic potentials. RNA interference (RNAi) was applied to down-regulate the ezrin expression in SF7721 and MHCC97-H. Changes of the cell growth and metastasis potentials after the RNAi treatment were studied. MTT assay was used to detect cell proliferation changes and Transwell assay was applied to observe the changes of cell motility and invasiveness.

RESULTSBoth ezrin and cytoskeleton protein were demonstrated in the cytoplasma of the cells at the same time. The expression of them in cell lines with high metastatic potential, such as SF7721, MHCC-1 and MHCC97-H was obviously higher than in those with low metastatic potentials, such as SMMC-7721, Hep3B and HepG2 (chi2= 13.277, P = 0.010; chi2= 21.815). The mRNA and ezrin and cytoskeleton protein gamma-actin were over-expressed in HCC cell lines with high metastatic potentials. The expressions of beta-actin of cell lines with different metastatic potentials showed no differences. Ezrin protein was successfully down-regulated and the proliferation and the invasiveness of the cells decreased with low ezrin protein level in SF7721 and MHCC97-H.

CONCLUSIONOver-expression of ezrin and cytoskeleton protein gamma-actin are associated with the process of metastasis of hepatocellular carcinoma cells. The growth and invasiveness of SF7721 and MHCC97-H cells can be inhibited by down-regulating ezrin expression.

Actins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cytoskeletal Proteins ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness

Objective:To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for re?pairing articular cartilage injury in vivo. Methods:Rabbit bone marrow mesenchymal stem cells(BMSCs)were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treat?ment. After animals anesthesia,a full thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile,the transfected cells were implanted to repair the rabbit model with full thickness cartilage defects. Cartilage defects tissue was observed with light microscope,electron microscope,HE and im?munohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation. Re?sults:At 3 days after the transfection,Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection,the expression of collagen typeⅡbegan and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox 9 gene. Histologi?cal observation showed that at 6 weeks after the operation,the defects in the experimental group was filled with hyaline like cartilage tissue,12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively,the defects were filled with fibrous tissues in control groups. Meanwhile,immunohistochemical stain?ing of sections with typeⅡcollagen antibodies showed the proteins in the regenerated tissue stained positive for typeⅡcolla?gen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances. Conclusion:Overexpression of Sox9 gene on rab? bit bone marrow mesenchymal stem cells(BMSCs)promote the repair of cartilage defect.

Objective: To observe the clinical effect of Modified Taohong Siwu Decoction (MTSD) in treating pediatric intractable nephropathy (PIN). Methods: Ninety-five patients with PIN were divided into 2 groups at random. The 60 patients in the treated group were treated with MTSD and the 35 patients in the control group were treated with heparin. The clinical therapeutic effect and levels of thromboxane B2 (TXB2) and 6-keto-prostacyclin F1α (6-keto-PGF1α) before and after treatment were observed. Results: The total effective rate in the treated group was 81.7%, which was similar to that in the control group (80.0%, P>0.05). The levels of TXB2 and TXB2/6-keto-PGF1α ratio were higher in both groups of the patients as compared with those of the healthy control (P<0.01). After treatment, the two criteria were significantly improved in the two treated groups, as compared with those before treatment, the difference was significant (P<0.01), while in comparison between the MTSD treated group and the control group, no significant difference was found (P>0.05). Conclusion: MTSD has good effect in treating PIN, it could improve the metabolic unbalance of thromboxane and prostacyclin.

OBJECTIVETo extract tumor interstitial fluid (TIF) from MKN-45 gastric cancer which is similar to "muddy phlegm" in Chinese medicine and observe influences of MKN-45 tumor interstitial fluid (MKN-45 TIF) intervention on metastasis of gastric cancer and on the expressions of vascular endothelial growth factor (VEGF), kinase insert domain containing receptor (KDR), epithelial-cadherin (E-cad), cyclooxygenase-2 (COX-2), intercellular adhesion molecule-1 (ICAM-1) and telomerase genes and proteins in primary tumor tissue.

METHODSAn MKN-45 tumor-bearing model was established in 50 nude mice. The modeled animals were equally randomized to 5 groups: the simple tumor-bearing group (model group), the normal saline (NS) via tail vein injection (i.v.) group (NS i.v. group), MKN-45 TIF i.v. group (TIF i.v. group), NS intraperitoneal injection (i.p.) group (NS i.p. group), and MKN-45 TIF i.p. group (TIF i.p. group). The TIF and NS intervention groups received injection (i.p. or i.v.) of MKN-45 TIF or NS twice a week, 0.2 mL at a time. After 8 weeks, the primary tumors were removed, weighed and HE stained to observe tumor metastasis. The primary tumor tissues were analyzed by immunohistochemistry and real-time quantitative PCR to detect expressions of VEGF, KDR, E-cad, COX-2, ICAM-1, and telomerase genes and proteins in different groups.

RESULTSThere were significant differences in tumor weight between TIF intervention groups and the model and NS intervention groups. Tumor metastasis was observed in all 5 groups, but the tumor metastasis rate in TIF intervention groups was significantly higher than those in the model and NS intervention groups. The gene and protein expressions of gastric cancer-related factors VEGF, KDR, COX-2, ICAM-1 and telomerase were unregulated while the gene and protein expressions of E-cad were downregulated in TIF intervention groups.

CONCLUSIONSTIF promotes tumor growth, invasion and metastasis of gastric cancer. These findings provide preliminary experimental clues for verifying the hypothesis of "tumor-phlegm microenvironment".

Animals ; Cadherins ; genetics ; metabolism ; Cell Line, Tumor ; Cyclooxygenase 2 ; genetics ; metabolism ; Extracellular Fluid ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Stomach Neoplasms ; metabolism ; secondary ; Telomerase ; genetics ; metabolism ; Tumor Microenvironment ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

OBJECTIVETo reveal the relationship between the storage time of the bark of Magnolia officinalis and the content of phenols in it, and lay a theoretical foundation for the harvest, processing, management and storage.

METHODThe contents of magnolol and honokoiol in 15 bark samples, collected from the main producing areas in China, were determined in the time of freshly harvest and 3 and 10 years after respectively by HPLC method.

RESULTIt showed that within a certain period of time, bark storage was favorable to conversion and accumulation of phenols, that the content of magnolol tended to increase from year 0 to year 3, then followed by slight decrease with years on account of volatilization of phenols, but was still higher when the bark was stored for 10 years than that that when the bark was freshly harvested, and the content of honokoiol still tended to increase when the bark had been stored for 10 years.

CONCLUSIONThe phenols in bark of M. officinalis is quite stable and the bark can be stored for 10 years or longer.

Biphenyl Compounds ; analysis ; Drug Storage ; methods ; Drugs, Chinese Herbal ; chemistry ; Lignans ; analysis ; Magnolia ; chemistry ; Plant Bark ; chemistry ; Time Factors

OBJECTIVETo study the therapeutical effects of crossing anastomosis of nerve on the peripheral and central nerve injuries.

METHODSTwelve kinds of central and peripheral nerve disorders and their complications were treated with 11 kinds of crossing anastomosis of nerve bundles near the innervated organs. After nerve injury and repair, somatosensory evoked potentials (SEPs) and horseradish peroxidase (HRP) retrograde tracing studies were used to investigate the rabbit's nerve function and morphology.

RESULTSThe ulcers of all patients healed. Sensation, voluntary movement, and joint function recovered. Four weeks after the anastomosis of distal stump of radialis superficialis nerve and median nerve, pain sensation regained and SEPs appeared. HRP retrograde tracing studies demonstrated sensory nerve ending of medial nerve formed new connection with the body of neuron.

CONCLUSIONCrossing anastomosis of nerve is an effective method to treat peripheral and central nerve injuries.

Adolescent ; Adult ; Anastomosis, Surgical ; methods ; Animals ; Central Nervous System ; injuries ; surgery ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Neurosurgical Procedures ; methods ; Peripheral Nerve Injuries ; Peripheral Nerves ; surgery ; Rabbits ; Trauma, Nervous System ; surgery ; Young Adult

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics