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 study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients.

METHODSRecruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed.

RESULTSThe sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSInfection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

Anti-Bacterial Agents ; administration & dosage ; pharmacology ; therapeutic use ; Communicable Diseases ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Humans ; Infertility ; Infertility, Male ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; drug effects ; Urokinase-Type Plasminogen Activator ; metabolism

Aneurysm, Dissecting ; genetics ; Aortic Aneurysm ; genetics ; Diseases in Twins ; Humans ; Male ; Middle Aged

This paper reviews and summarizes the background，stages，tasks，achievements and experiences of the China's new round of healthcare reforms. Lessons gained from the reforms are valuable for deepening the healthcare reforms of China.

Abstract?AIM:To study the Influence and outcomes of eye healthcare information teaching for the visual development of children under“combination of medicine and education” in kindergartens.?METHODS:The children(5-6 years old) were randomly selected from 6 kindergartens in Nanjing, 3 in Yuhua District as the experimental group, and the other 3 in Jianye District as the control group.A one-year follow-up was conducted to evaluate the difference of visual development, including the rate of low vision, rate of referral caused by refractive abnormality, rate of astigmatism, the average of the equivalent spherical lens and the rate of lacking physiological hypermetropia, between the experimental group and the control group children.?RESULTS:One school year later,the rate of low vision, rate of referral caused by refractive abnormality and the rate of astigmatism, were significantly lower (P<0.05)in the experimental group under the mode of“combination of medicine and education”, which were also lower than those before experiment(P<0.05).The average of the equivalent spherical lens of experimental group increased and the rate of lacking physiological hypermetropia decreased significantly, compared with the control group ( P <0.05 ) and with those before experiment(P<0.05).?CONCLUSION:Using “combination of medicine and education” eye health care model,is good for children's visual development,so as to reduce the rate of low vision and delay the occurrence of myopia.

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

To investigate the chemical constituents of the whole plants of Bidens bipinnata, the separation and purification of constituents were performed by chromatography on macroporous resin, silica gel, MCI and Sephadex LH-20. Their structures were elucidated by spectroscopic data as quercetin (1), quercetin-3-0-alpha-L-rhamnoside (2), keampferol-3-O-beta-D-glucopyranoside (3), keampferol-3-O-alpha-L-rhamnoside (4), 3', 5-dyhydroxy-3, 6, 4'-trimethoxyl -7-O-beta-D-glucopyranoside flavonoid (5), 7, 8, 3', 4'-tetraflavanone(6), (2S)- and (2R)-isookanin-7-O-beta-D- glucopyranoside (7a/7b), (2S)- and (2R)-3'-methoxy-isookanin-8-O-beta-D-glucopyranoside (8a/8b), 6, 7, 3', 4'-tetrahydroxyaurone(9), maritimetin (10), esculetin (11), 3-O-caffeoyl-2-methyl-d-erythrono-1, 4-lactone (12), (7S, 8R) balanophonin-4-O-beta-D-glucopyranoside (13), eugenyl-O-beta-apiofuranosyl-( 1"-6') -O-beta-glucopyranoside (14), and (+)-syringaresinol-4'-O-beta-D-glucopyranoside (15). Compounds 8, 13, 14, and 15 were isolated from this genus for the first time. Compounds 1 and 6 were potent inhibitors against HSC-T6 cells in vitro and compounds 1, 2, 6, and 7 were capable of decreasing the inflammatory cytokine production of macrophage cells in vitro.
Bidens ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

This experiment was expected to test whether nitric oxide (NO) exerted significant effect on the central respiratory rhythm. Experiments were performed on in vitro brainstem slice preparations from neonatal rats. These preparations include the medial region of the nucleus retrofacialis (mNRF); a part of pre-Bötinger complex, ventral respiratory group (VRG) and dorsal respiratory group (DRG). Respiratory-related burst activities were recorded from hypoglossal nerve rootlets before and during superfusion of the slice preparation with L-Arginine (L-Arg), sodium nitroprusside (SNP) or 7-nitro indazole (7-NI, an inhibitor of NO synthase). After perfusion with L-Arg and SNP, there was no significant change in respiratory rhythmical discharge activity (RRDA), but 7-NI decreased the integral amplitude of burst and inspiratory time. These results indicate that NO may take part in the inspiratory off-switching mechanism and that it also modulates the amplitude of respiratory-related bursts.
Animals ; Animals, Newborn ; Arginine ; pharmacology ; Brain Stem ; physiology ; Electrophysiology ; Indazoles ; Neurons ; physiology ; Nitric Oxide ; physiology ; Nitric Oxide Synthase ; antagonists & inhibitors ; Nitroprusside ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiration ; Respiratory Center ; physiology

BACKGROUNDConnective tissue growth factor (CTGF) is a secreted protein containing several domains that mediate interactions with growth factors, integrins and extracellular matrix components. CTGF plays an important role in extracellular matrix production by its ability to mediate collagen deposition during wound healing. CTGF also induces neovascularization in vitro, suggesting a role in angiogenesis in vivo. We herein evaluated whether CTGF was required for extracellular matrix synthesis of meniscal fibrochondrocytes and/or angiogenesis during the repair of meniscal tears.

METHODSMeniscal fibrochondrocytes were isolated from the inner-1/2 of rabbit meniscus by trypsin collagenase treatment and further treated with 100 ng/ml CTGF in vitro. Characterization of fibrochondrocytes was identified by flow cytometry analyzing CD31, CD44, CD45 and CD105, and was further tested by type II collagen immunocytochemistry. Changes in gene expression of meniscal fibrochondrocytes were monitored by quantitative real-time polymerase chain reaction. Histological sections prepared from a 3-mm portion of a longitudinal tearing defect in the middle of the rabbit meniscus were subjected to fluorescence-immunohistochemistry analysis at 1, 4 and 10 weeks following surgical treatment with 1.5 µg of CTGF/fibrin-glue composites.

RESULTSQuantitative RT-PCR assay showed that types I and II collagen and vascular endothelial growth factor mRNA expression in the 100 ng/ml CTGF group were remarkably enhanced as compared to levels in the no-dose group at 14 days ((2.38 ± 0.63) fold, (2.96 ± 0.87) fold, (2.14 ± 0.56) fold, respectively). Likewise, fluorescence-immunohistochemical analysis revealed that in the group implanted with CTGF-fibrin glue, types I and II collagen, as well as the capillaries, completely filled the defect by 10 weeks, postoperatively. In contrast, only soft tissue repair occurred when PBS-fibrin glue was implanted.

CONCLUSIONSThese findings suggest that CTGF can significantly promote extracellular matrix deposition (types I and II collagen) within the meniscal avascular zone; CTGF can greatly heighten the expression of vascular endothelial growth factor activity simultaneously in vivo, further enhancing the repair of meniscal tears in the avascular zone.

Animals ; Cells, Cultured ; Chondrocytes ; cytology ; Collagen Type I ; metabolism ; Collagen Type II ; metabolism ; Connective Tissue Growth Factor ; pharmacology ; Flow Cytometry ; Immunohistochemistry ; Male ; Menisci, Tibial ; cytology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Tibial Meniscus Injuries ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing ; drug effects

BACKGROUNDElevated levels of interleukin-7 (IL-7) have been correlated with CD4(+) T cell depletion and the emergence of syncytium-inducing (SI) variants in human immunodeficiency virus type-1 (HIV-1) infection, and suggested as an indicator of acquired immunodeficiency syndrome (AIDS) disease progression. Therefore, we investigated the effects of IL-7 on disease progression and virus phenotype in Chinese HIV/AIDS patients.

METHODSIn a cross-sectional study of 71 untreated HIV-1 seropositive individuals and 12 healthy donors, plasma IL-7 levels were determined by an ultra sensitive enzyme-linked immunosorbent assay (ELISA), and its relations to CD4(+) T cells, CD8(+) T cells, plasma viral loads and HIV phenotypes were analyzed.

RESULTSSignificant higher IL-7 levels were found in Chinese HIV/AIDS patients [(3.33 +/- 3.60) pg/ml] than those of health controls [(1.2 +/- 0.81) pg/ml] (P < 0.05), and IL-7 levels were inversely associated with CD4(+) T cell counts (r = -0.497, P < 0.01). Furthermore, IL-7 levels were significant higher in patients with SI variants [(9.12 +/- 4.55) pg/ml] than those with non-syncytium-inducing variants [(1.50 +/- 2.69) pg/ml] (P < 0.01).

CONCLUSIONSIncreased IL-7 levels were found in Chinese HIV/AIDS patients and significantly associated with disease progression, thus increased IL-7 plasma levels may indicate disease progression.

Adult ; CD4 Lymphocyte Count ; Disease Progression ; HIV Seropositivity ; blood ; immunology ; virology ; HIV-1 ; Humans ; Interleukin-7 ; blood ; physiology ; Phenotype ; Viral Load