The study was aimed to evaluate if the modified in situ cryopreservation could affect the biological function of mesenchymal stem cells (MSC) in vitro. Mesenchymal stem cells from human bone marrow were isolated by standard method and characterized with their morphology, cell-surface antigen profile and differentiation repertoire in vitro. The culture-expanded MSC were cryopreserved in situ with culture medium (DMEM-LG) containing 10% D MSO and 30% selected FCS in -70 degrees C. Following recovery of cryopreservation, differentiation to adipocytes, chondrocytes, and osteoblast in vitro and cell cycle analysis were performed to investigate whether the cryopreservation would change the differentiation potential of MSC. The results showed that after recovery of cryopreservation, there was no changes detected as compared with the culture-expanded MSC in both differentiation potency and growth pattern at 12 weeks. In conclusions: this optimized short term in situ cryopreservation at -70 degrees C could retain biological characteristics of human MSC for at least 3 months, and this method may be useful for cryopreservation of hum an bone marrow MSCs.
Bone Marrow Cells ; cytology ; Cell Cycle ; Cell Differentiation ; Cell Separation ; Cell Survival ; Cryopreservation ; Humans ; Mesenchymal Stromal Cells ; cytology

OBJECTIVETo study the effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer.

METHODSIn vitro, the inhibitory effects of YH-16 and 5-FU on the growth of vascular endothelial cells and colorectal cancer cells were examined by MTT assay. In vivo, colorectal cancer cells were transplanted into BALB/c mice, and the mice were divided into six groups randomly:control group, low-dose YH-16 group, middle-dose YH-16 group, high-dose YH-16 group, 5-FU group and combination group. The number of liver metastases, the size of primary tumor and the toxicity were examined after 2 weeks postoperatively. The expression of vascular endothelial growth factor (VEGF) in liver metastases was detected by immunohistochemistry, and tumor microvessel density (MVD) was measured by immunostaining with CD34 and factor VIII (monoclonal antibodies.

RESULTSIn vitro, YH-16 inhibited the growth of colon cancer cells and vascular endothelial cells, with the IC50 at (2.16+/-0.28) microg/ml and (0.64+/-0.10) microg/ml respectively. In vivo high-dose YH-16 and 5-FU had a remarkable inhibitory effect on liver metastasis, and the combination group showed significant enhancement on this effect (P< 0.05). The combination group and 5-FU group could inhibit the growth of primary tumor, but not found in YH-16 group. The toxicity of YH-16 was lower than that of 5-FU (P< 0.05), and the difference was not found in the toxicity between combination group and 5-FU group (P > 0.05). Expression of VEGF in liver metastases was clearly inhibited by YH-16 in combination with 5-FU or 5-FU alone compared to the control group, and MVD in middle-dose and high-dose YH-16 group, 5-FU group and combination group was lower than that in control group (P< 0.05).

CONCLUSIONSThe angiogenesis inhibitor YH-16 can inhibit liver metastasis of colorectal cancer through inhibiting the growth of vascular endothelial cells. YH-16 in combination with 5-FU has additive effect on inhibitory activity against liver metastasis.

Angiogenesis Inhibitors ; therapeutic use ; Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; drug therapy ; pathology ; Drug Therapy, Combination ; Female ; Fluorouracil ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the prevalence of psychiatric disorders in a representative sample of primary and middle school students in Hunan Province.

METHODSA total of 9 495 children aged 5-17 years from Hunan urban and rural schools were enrolled by a cluster sampling and a two-phase design. The students' psychiatric status was assessed using the Investigation Screening Inventory for Child Mental Disorder and a semi-structured interview designed based on the DSM-IV criteria.

RESULTSThe overall prevalence of psychiatric disorders was 16.22%. Attention-deficit and disruptive behavior disorders were the commonest in the diagnostic categories of psychiatric disorders (10.69%). Regarding specific disorders, the most prevalent was attention-deficit/hyperactivity disorder (5.95%). Psychiatric disorders were more prevalent in boys than in girls (20.49% vs 11.16%; p<0.01). The prevalence of attention-deficit and disruptive behavior disorders in boys was higher than in girls (14.76% vs 5.87%; p<0.01). The prevalence of psychiatric disorders in middle school students (12-17 years) was significantly higher than in primary students (5-11 years) (18.38% vs 14.64%; p<0.01). There were no significant differences in the prevalence of psychiatric disorders between urban and rural students.

CONCLUSIONSPsychiatric disorders are common among primary and middle school students in Hunan Province. The prevalence of this disorder in boys is higher than in girls. The middle school students have higher prevalence than primary students.

Adolescent ; Age Factors ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Mental Disorders ; epidemiology ; Prevalence ; Sex Factors

OBJECTIVETo detect the expression and correlation of cyclooxygenase 2 (COX-2)and key enzymes both of mitogen activated protein kinase (MAPK) and nuclear factor-kappa B( NF-KB) pathways in middle turbinate mucosa of chronic rhinosinusitis (CRS) and to investigate their roles in CRS pathogenesis.

METHODSTwenty-four lateral mucosa of CRS middle turbinates were equally divided into 3 groups according to FESS-97 Haikou criterion (CRS type I stage 2 in group 1, type II stage 2 in group 2, and type III in group 3), and 8 normal mucosa were enrolled as control. Immunohistochemistry and fluorescent real time quantitative polymerase chain reaction (FQ-RT-PCR) were performed to detect the expression of COX-2, ERK, p38MAPK, JNK and NF-kappaB subunits. The correlations between cox-2 and MAPK, NF-kappaB pathway were statistically treated by Pearson test.

RESULTSPositive expressions of COX-2, ERK, p38MAPK and NF-kappaB subunits were detected in CRS groups, which were stronger than those in control group, by immunohistochemistry and FQ-RT-PCR (P < 0.05). Statistic difference was not found among CRS groups (P > 0.05). Negative expression of JNK was detected in all groups. Significantly positive correlation between protein and RNA expression of COX-2,ERK,p38MAPK and NF-kappaB subunits in each CRS group was confirmed by Pearson correlation treatment (P < 0.05). Significantly positive correlation of protein and RNA expression between COX-2 and ERK, p38MAPK in same CRS group was also founded (P < 0.05). The expression of COX-2 and the nucleic expression of NF-kappaB subunits in same CRS group was proved to be positively correlated (P < 0.05).

CONCLUSIONSUpregulated expression of COX-2 is correlated with upstream ERK, p38MAPK and NF-kappaB pathway in CRS. It indicates the involvement of ERK, p38MAPK and NF-kappaB signal transduction pathway in regulation of COX-2 in CRS.

Adolescent ; Adult ; Case-Control Studies ; Chronic Disease ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Mitogen-Activated Protein Kinases ; metabolism ; NF-kappa B ; metabolism ; Nasal Mucosa ; metabolism ; Signal Transduction ; Sinusitis ; metabolism ; Turbinates ; metabolism ; Young Adult

OBJECTIVETo summarize the management of anastomotic leak following surgery for esophageal carcinoma.

METHODSThe medical records of the patients developing digestive tract leak after surgery for esophageal carcinoma in our hospital from January 2003 to March 2011 were retrospectively analyzed.

RESULTSA total of 36 patients were included, in whom 13 developed cervical anastomotic leak, 18 had intra-thoracic anastomotic leak, and 5 had intra-thoracic gastric necrosis. Of these patients, 7 were treated with resurgery, 6 with esophageal stent implantation, and 23 with conservative treatment. Treatment lasted for 5 to 181 days, averagely 47.0 +/- 31.9 days. After management, 9 patients died (25.0%). Among seven patients with resurgery, four had deceased, two were cured, and one developed leak again and was switched to conservative treatment until discharged. All the 6 patients treated with stent implantation were cured. Of the 24 patients receiving conservative treatment (including one switched from resurgery), 18 (75.0%) were cured and 1 was not cured but survived.

CONCLUSIONSAnastomotic leak following surgery for esophageal carcinoma should be treated individually based on the onset time, location, size, and extent of the leakage. Conservative treatment is still a safe and effective method. The efficacy of stent implantation needs further investigation to confirm.

Adult ; Aged ; Anastomotic Leak ; therapy ; Esophageal Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Precision Medicine ; Treatment Outcome

OBJECTIVETo investigate the protective effect of Astragalus Membranaceus Injection on human umbilical vein endothelial cells (HUVECs) against tumor necrosis factor alpha (TNF-alpha).

METHODSCultured passage 2 HUVECs were stimulated with TNF-alpha with or without a 2-h Astragalus Membranaceus Injection treatment. The expression of nuclear factor-kappaB (NF-kappaB) subunit p65 were evaluated by immuncytochemistical method, and the levels of p65 in the nuclei and the protein Ikappabetaalpha in the cytoplasm were evaluated by Western blotting. The levels of interleukin-6 (IL-6) and soluble intracellular adhesion molecule-1 (sICAM-1) in the cell culture were determined with ELISA.

RESULTSTNF-alpha induced the activation of NF-kappaB and increased the expressions of IL-6 and sICAM-1 in HUVECs. The activation of NF-kappabeta by TNF-alpha was suppressed by Astragalus Membranaceus Injection in a dose-dependent manner.

CONCLUSIONAstragalus Membranaceus Injection can inhibit the TNF-alpha-induced expression of IL-6 and sICAM-1 by suppressing NF-kappabeta activation, suggesting its protective effect on the endothelial function.

Astragalus membranaceus ; chemistry ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-6 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; pharmacology ; Umbilical Veins ; cytology

AIM To investigate the effect and mechanism of total anthraquinone extract of Rhei Radix et Rhizoma on cerebral ischemia-reperfusion injury and to provide relevant data references for its promising use in the management of cerebral ischemia-reperfusion injury.METHODS SD rats randomly assigned to model group,sham operation group,nimodipine group,total anthraquinone extract groups (high,medium and low dose),8 in each group,were orally administered with corresponding drugs daily for a week,with rats of the model group and sham operation group given the same volume of normal saline before the models established by middle cerebral artery occlusion (MCAO).MCAO started thirty minutes after final oral administration,and the induced ischemia went on for 1.5 h for a further reperfusion,24 h after which the neurological function score,brain index,brain water content and cerebral infarct volume were measured.Elisa kits were used to detect superoxide dismutase (SOD),Malondialdehyde (MDA),Nitric oxide (NO),interleukin-1 β (IL-1β),tumor necrosis factor (TNF),interleukin-6 (IL-6).RESULTS The total anthraquinone extract of Rhei Radix et Rhizoma significantly improved the neurological function score,decreased the brain index,brain water content,reduced the cerebral infarct volume (P ＜ 0.05),increased the activity of SOD in brain tissue (P ＜ 0.01),and reduced the levels of MDA and NO in brain tissue (P ＜0.01),and the levels of IL-6,TNF and IL-1β in serum (P ＜0.01) as well.CONCLUSION The obviously protective effect on rats' cerebral ischemia-reperfusion injury by total anthraquinone extract of Rhei Radix et Rhizoma may contribute to its inhibition of inflammatory response,and its existence as an antioxidant as well.

OBJECTIVETo construct an eukaryotic expression vector for PRL-2 and evaluate its effect on the invasiveness and migration of a human hepatocellular carcinoma cell line.

METHODSRT-PCR was performed to amplify the complete PRL-2 open reading frame using the total mRNA of hepatocellular carcinoma HepG2 cells as the template. PRL-2 gene was inserted into the pGEM T easy vector and sequenced, and the correct PRL-2 sequence was subcloned into the mammalian expression vector pcDNA3.1+. The constructed PRL-2 vector was transfected into CL1 cells via lipofectamine, and the stable expression of PRL-2 mRNA was detected by RT-PCR, the expressed protein identified by immunohistochemistry and Western blotting, and the effect of PRL-2 on the adhesion ability of CL-1 cell evaluated with MTT assay 20 and 120 min after transfection. The effect of PRL-2 on the invasive migration of CL-2 cells was evaluated according to the number of cells penetrating the Matrigel layer of polycarbonate membrane of Boyden chamber.

RESULTSRT-PCR yielded a fragment of 504 bp and the inserted PRL-2 sequence was verified by sequence analysis. The subclones were identified by restriction endonuclease digestion, and a G418-resistant clone, PRL-2-CL1, was obtained after 8 weeks of selection. RT-PCR showed stable expression of PRL-2 mRNA, and Western blotting confirmed overexpression of PRL-2 protein in the transfected cells. PRL-2 increased the adhesion rate of CL-1 cells to fibronectin at 20 min and 120 min after transfection (P<0.05), and also the number of CL-1 cells penetrating the polycarbonate membrane from 10.0+/-3.7 to 44.8+/-2.6 (P<0.05).

CONCLUSIONAn eukaryotic expression vector of PRL-2 has been successfully constructed, which allows stable and efficient expression in CL-1 cell line. PRL-2 can promote cell adhesion and invasion activity of human hepatocellular carcinoma cells.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Adhesion ; genetics ; Cell Line, Tumor ; Cell Movement ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; genetics ; Protein Tyrosine Phosphatases ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo study the effect of skin-derived progenitor cell (SKP) combined with hyaluronic acid( HA) on the wound healing in diabetic rats.

METHODSSKP of Spraque-Dawley (SD) neonate rats were isolated and cultured and mixed with HA. The differentiation characteristics of SKP in the culture were observed. Sixty SD rats were injected intraperitoneally with 65 mg/kg streptozotocin( STZ) to induce diabetes. Two symmetrical full-thickness cutaneous wounds( 1.0 cm in diameter) were made on the back of each SD rat and randomly divided into A (n = 20, with treatment of 100 mircol SKP-HA) , B (n = 20, with treatment of 100 mirol HA) , and C ( n = 20, with treatment of DMEM/F12 culture medium) groups. Tissue samples from wound in each group were harvested on 1, 2, 3, 4 weeks after the treatment. Wound healing rate, changes in histomorphology, the content of hydroxyproline ( HYP) , and immigration of labelled SKP were determined and analyzed.

RESULTSSKP grew well when cultured with HA. The characteristics of SKP to differentiate into lipocyte, neuron, and neurogliocyte remained in the culture. Compared with that in C group, epithelization in the wounds of A and B groups appeared earlier. The wound healing rate in A group [ (72.1 +/- 2. 8)% ] and B group [ (53.7 +/- 2. 9)% ] were obviously higher at 2 post-treatment weeks(PTW) than that in group C [(42. 5 +/- 1.5)% ( P <0.05) , and that in A group was obviously higher compared with B and C groups at 3 PTW ( P < 0. 05 or 0. 01). The wound healing rates in A and B groups were (100. 00 +/- 0.00) % at 4 PIW, which were obviously higher than that of group C( P <0.01) . There was no obvious difference in the HYP content among the 3 groups at 1 PIW ( P > 0. 05) , but it was obviously higher in A and B groups than that in C group at 2,3,4 PTW( P <0.01) , and that in A group was significantly higher than that in B group at 3 and 4 PTW( P <0. 01). SKP survived well on the wound, and migrated towards the dermis as time elapses.

CONCLUSIONSKP-HA composition can promote wound healing in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Hyaluronic Acid ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; cytology ; Stem Cells ; chemistry ; cytology ; Wound Healing ; drug effects

OBJECTIVETo investigate the association of 86 bp variable number tandem repeat (VNTR) polymorphism of interleukin-1 receptor antagonist (IL-1Ra) gene with lumbar disc disease and intervertebral disc degeneration.

METHODSThe 86 bp VNTR polymorphism of IL-1Ra gene was analyzed with PCR and electrophoresis for 81 patients with lumbar disc disease and 101 volunteers without sciatica (control). The grade of intervertebral disc degeneration was determined with magnetic resonance imaging, and the association of 86 bp VNTR polymorphisms with lumbar disc disease and intervertebral disc degeneration in those younger than 45 years was assessed.

RESULTSThe presence of 86bp VNTR polymorphisms of IL-1Ra gene was detected in both patients with lumbar disc disease and the control subjects. The distribution of 86 bp VNTR polymorphisms of IL-1Ra gene showed no significant difference between the two groups, but the distributions of 1/1, 1/2 and 2/2 or 1, 2 genotypes differed significantly. The current data did not support a significant association between the distribution of IL-1Ra gene 86bp VNTR polymorphism and lumbar disc degeneration.

CONCLUSIONSIL-1Ra gene 86bp VNTR polymorphism is present among Chinese population in association with lumbar disc disease, but not with lumbar disc degeneration.

Adult ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Female ; Genome-Wide Association Study ; Humans ; Interleukin 1 Receptor Antagonist Protein ; genetics ; Intervertebral Disc Degeneration ; genetics ; Male ; Middle Aged ; Minisatellite Repeats ; Polymorphism, Genetic ; Spinal Diseases ; genetics ; Young Adult