Objective To observe the survival, migration and differentiation of adipose-derived adult stem (ADAS) cells and the recovery of motor function of ischemic rats after ADAS cell transplantation. Methods Eighteen adult male SD rats, weighted about 250-300 g, were chosen and received left middle cerebral artery occlusion (MCAO) operation. And then, they were equally randomized into untreated group, control group and ADAS cell transplantation group. Tail vein injection of Dulbecco's modified eagles medium (DMEM) and ADAS cells were performed in the control group and ADAS cell transplantation group 3 h after MCAO, respectively. These animals were euthanized 14 d after MCAO. Immunofluorescence for BrdU, NSE, MAP-2 and GFAP were processed to identify the survival, migration and differentiation of grafted ADAS cells in the brain, and at the same time, scores of neurological deficit scale were used to assess the improvement of motor function. Results After the transplantation, numerous ADAS cells labeled with BrdU were presented in the ischemic points and surrounding areas. A few BrdU/GFAP, BrdU/NSE and BrdU/MAP-2 -positive cells were noted in the ischemic points of ADAS transplantation group 14 d after MCAO. The neurological functional recovery in the ADAS cell transplantation group was significantly improved as compared with that in the control group 14 d after MCAO (P＜0.05). Conclusion ADAS cells can migrate into the ischemic hemisphere and differentiate into neuron-like and astrocytic-like cells after the injection by venous approach in the rat models with MCAO. The intravenous administration of ADAS cells into rats with MCAO leads to good functional outcome and few lesion sizes.

OBJECTIVETo estimate the benchmark dose for osteoporosis caused by cadmium exposure in a Chinese general population with an epidemiological study.

METHODSThe inhabitants living in both cadmium polluted and non-polluted areas served as the exposure group and the control group. Urinary cadmium (UCd) and Blood cadmium (BCd) were used as exposure biomarkers while the Z score was used as effect biomarker for the osteoporosis.

RESULTSThe UCd and BCd in the habitants of the polluted areas were significantly higher than those in the habitants of the control area on average (P < 0.05) and the UCd and BCd in the habitants of the highly polluted areas were significantly higher than those in the habitants of the moderately polluted area on average (P < 0.05). The bone mineral density was significantly decreased in the groups of the highest UCd and BCd level compared with the 5 microg/g Cr group with the significant difference (P < 0.05). The morbidity of the osteoporosis would increase significantly with the increase of the cadmium exposure (P < 0.05) with the linear correlation (P < 0.05). BMDs were calculated using BMDS Version l.3.2 software and BMDLs were also determined. The BMDL of UCd for cadmium-induced osteoporosis was higher than those representing cadmium-induced renal dysfunction.

CONCLUSIONHigh level of cadmium exposure can induce osteoporosis, which occurs later than renal damage related to cadmium exposure. The BMD is a practical method.

Aged ; Bone Density ; drug effects ; Cadmium ; adverse effects ; metabolism ; China ; epidemiology ; Dose-Response Relationship, Drug ; Environmental Exposure ; Female ; Humans ; Male ; Middle Aged ; Osteoporosis ; chemically induced ; epidemiology

OBJECTIVETo investigate the relationship between perinatal risk factors such as premature, low birth weight, small for gestational age and childhood cerebral palsy (CP).

METHODSA cross sectional survey was carried out among 305,263 children aged 1 - 6 years old in seven cities of Jiangsu Province, China from May to July 1997. The perinatal risk factors were analysed.

RESULTSFour hundred and eighty-four cases of CP were found among this population. The prevalence of CP for children aged 1 - 6 years old was 1.59 per thousand. The prevalence of CP were strongly correlated to prematurity (RR = 25.16), low birth weight (RR = 19.63), and also highly correlated to small for gestational age (RR = 4.34). For smaller groups divided by small for gestational age (SGA), appropriate for gestational age (AGA), large for gestational age (LGA) and then by gestational age, prematurity was found to be at high risk in SGA (RR = 9.29), AGA (RR = 28.34) and LGA (RR = 21.41) groups. For groups divided by gestational age and then by SGA, AGA and LGA, SGA was found to have significantly high risk in premature (RR = 1.45), mature (RR = 4.41) and postmature (RR = 3.19) groups. Nine groups were divided by the gestational age along with SGA, AGA and LGA, rates of CP were found to be significantly higher in most groups than in the term AGA group. Compared with the rate of CP in the term AGA group, the RR were calculated and showed as followings (from higher to lower), premature SGA (RR = 40.99), premature AGA (RR = 28.34), premature LGA (RR = 21.08), postmature SGA (RR = 8.39), mature SGA (RR = 4.41) and postmature AGA (RR = 2.63).

CONCLUSIONPrematurity and small for gestational age are both independent risk factors for cerebral palsy. Postmaturity and large for gestational age are not risk factors.

Cerebral Palsy ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Gestational Age ; Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Infant, Premature ; Male ; Risk Factors

AIM To establish a ultra-high performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS) method for the simultaneous content determination of notoginsenoside R1,ginsenoside Re,ginsenoside Rg1,ginsenoside Rb1,ginsenoside Rd,taurine,taurocholic acid,cholic acid,glycocholic acid,glycodeoxycholicacid,chenodeoxycholic acid,deoxycholic acid and muscone in Pientzehuang (Bovis Calculus,Moschus,Notoginseng Radix et Rhizoma,etc.).METHODS The analysis of methanol extract of this drug was performed on a 40 ℃ thermostatic Waters CORTECS UPLC C18column (100 mm × 2.1 mm,1.6 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 0.25 mL/min in a gradient elution manner.RESULTS Thirteen constituents showed good linear relationships within their own ranges (r ＞0.998 5),whose average recoveries were 91.1％-105.3％ with the RSDs of 2.4％-4.6％.CONCLUSION This accurate,simple,sensitive and reproducible method can be used for the quality control of Pientzehuang.

OBJECTIVETo explore the influence of beta-elemene on the proliferation, migration and RhoA expression of hepatic stellate cells (HSC) induced by angiotensin II (ANG II).

METHODSHSC were incubated in vitro. Proliferation and migration of the HSC were induced by ANG II. The effect on the proliferation of HSC was determined by MTT colorimetry. The migration ability was detected by transwell chamber cultures. Total RNA was extracted by TRizol reagent and gene levels were determined by semi-quantitative RT-PCR. Protein levels were determined by Western blot.

RESULTSDifferent concentrations (from 1 to 10 micromol/L) of ANG II markedly promoted the growth of the HSC in a concentration dependent way (0 micromol/L ANG II, F = 112.640, P less than 0.01). 10, 8, 4 micromol/L ANGII significantly induced HSC migration, F = 117.496, P less than 0.01. Compared with the 4 micromol/L ANG II group, 10 mg/L, 5 mg/L, 2.5 mg/L beta-elemene markedly inhibited HSC proliferation and migration induced by 4 micromol/L ANG II (F values were 95.706 and 55.600 and P less than 0.01). 4 micromol/L ANG II markedly promoted the protein and mRNA expressions of RhoA in HSC. 10 mg/L, 5 mg/L and 2.5 mg/L beta-elemene notably inhibited the expressions of RhoA protein and mRNA (F values were 217.119 and 18.010).

CONCLUSIONANG II can significantly induce the proliferation and migration of HSC. Beta-elemene can inhibit the proliferation and migration of HSC induced by ANG II. The effects of beta-elemene are mediated through inhibiting the RhoA signal transduction pathway and are associated with RhoA.

Angiotensin II ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hepatic Stellate Cells ; cytology ; drug effects ; Humans ; Sesquiterpenes ; pharmacology ; rhoA GTP-Binding Protein ; metabolism

OBJECTIVEIbandronate, a third generation bisphosphonate, inhibits bone resorption in human and animal studies. This study is to evaluate the efficacy and safety of ibandronate as a single agent in patients with tumor-associated hypercalcemia.

METHODSAn open, multicenter, non-controlled clinical trial was conducted in 22 patients. The patients received 2 mg ibandronate intravenously if the corrected calcium was less than 3.0 mmol/L but more than 2.7 mmol/L; they received 4 mg ibandronate iv if corrected calcium was more than 3.0 mmol/L.

RESULTSThere was 100% efficacy in these two dose groups but the calcium correcting effect was more pronounced in the 4-mg dose group than the 2-mg dose group. The most common adverse reactions were fever and skin itching with an incidence of 4.5%.

CONCLUSIONIbandronate is active in patients with tumor-associated hypercalcemia and the adverse effects are well tolerated.

Bone Neoplasms ; complications ; secretion ; Breast Neoplasms ; complications ; pathology ; Calcium ; blood ; Diphosphonates ; administration & dosage ; therapeutic use ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Female ; Fever ; chemically induced ; Humans ; Hypercalcemia ; blood ; drug therapy ; etiology ; Lung Neoplasms ; complications ; pathology ; Male ; Middle Aged ; Multiple Myeloma ; complications ; Phosphorus ; blood ; Pruritus ; chemically induced

OBJECTIVESTo investigate HEV infection in swine and the genotype relationship between swine and human HEV.

METHODSAnti-HEV IgG antibody was detected in the sera of swine using enzyme linked immunoassay (EIA), and HEV RNA was amplified by reverse transcription nested polymerase chain reaction (RT-nPCR). The Vector NTI Suite 7 and TreeView softwares were used for nucleotide sequences phylogenetic analysis of HEV isolated from human and swine.

RESULTSThe anti-HEV IgG positive rate was 16.67% (18/108). Among the 18 anti-HEV IgG positive sera, 2 sequences (11.11%, called S18 and S43, respectively) of HEV ORF1 (102-387bp) were amplified, with the identity of 99% between them. They had 76% to 77%, 78%, 76% to 79%, 85% to 86%, 77%, 80%, 79% and 75% - 79% homology at the nucleotide level with human HEV genotypes 1 to 8, respectively. One (S18) of them was also amplified out in ORF2 region (5,994-6 297bp) and showed 76% to 78%, 74%, 74% to 77%, and 85% to 94% identity with human HEV genotypes 1 to 4 at the nucleotide level, respectively.

CONCLUSIONHEV sequences isolated from swine belong to human HEV genotype 4.

Animals ; Antibodies, Viral ; blood ; Base Sequence ; Hepatitis E ; transmission ; veterinary ; Hepatitis E virus ; classification ; genetics ; Immunoglobulin G ; blood ; Molecular Sequence Data ; RNA, Viral ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Swine ; Swine Diseases ; virology

OBJECTIVETo construct a MYH9 gene knockout model in MGC803 cell line using transcription activator-like effector nuclease (TALEN) and observe its effect on cell cycle and apoptosis.

METHODSAccording to FastTALE(TM) TALEN Kit, we designed TALEN pairs and constructed the plasmids targeting to MYH9 gene. After detecting their activity in MGC803 cells by plasmid transfection, DNA sequencing, RT-PCR and western blot, we selected the monoclonal cells and studied the changes in the cell cycle and apoptosis.

RESULTSMYH9 gene could not be knocked out but knocked down in selected MGC803 monoclonal cells, which caused cell cycle arrested at G2/M phase (P<0.05) and a significant increase in the cell number with early apoptosis (P<0.01).

CONCLUSIONWe successfully generated a MYH9 knockdown model in MGC803 cell lines by TALEN, which could be in favor of MYH9 function study in gastric cancer.

Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Gene Knockdown Techniques ; Humans ; Molecular Motor Proteins ; genetics ; Myosin Heavy Chains ; genetics ; Plasmids ; Stomach Neoplasms ; Transfection

Objective Feasibility of using MNA cell-culture inoculation test to detect and isolate the street rabies virus. Methods Using MNA cell-culture inoculation test, fluorescent antibody test (FAT) and sandwich ELISA with double-antibodies to detect 33 specimens of street rabies virus, 20 specimens of negative canine brains and 4 specimens of healthy mice brains. Results 33 specimens of street rabies virus were positive to the cell-culture inoculation test but the others were negative. The concordances of MNA cell-cultured inoculation test with FAT and sandwich ELISA with double-antibodies were both 100%. Conclusion MNA cell-culture inoculation test appeared to be both highly sensitive and specific in detecting the street rabies virus, and could be used in detection and isolation of the virus.

OBJECTIVETo identify the gene mutation for two Chinese families with autosomal dominant non-syndromic hearing impairment(NSHI).

METHODSTwo NSHI pedigrees with common ancestor were identified by clinical examination and family investigation. Linkage analysis was performed for all known NSHI loci, and all exons and exon-intron boundaries of the non-muscle myosin heavy chain 14 (MYH14) gene were amplified by PCR and sequenced.

RESULTSThe disease-causing gene of these 2 pedigrees was fine mapped to the DFNA4 locus on 19q13.33. A heterozygous transition of c. 359T>C (p.S120L) in MYH14 gene was identified. The mutation was detected in all patients but not in normal members in the two families.

CONCLUSIONIt is the first report that mutation in MYH14 gene can cause dominant non-syndromic hearing impairment in Asian population, suggesting that MYH14 gene can be a disease-causing gene of Chinese patients with hearing impairment.

Female ; Hearing Loss ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Mutation ; Myosin Heavy Chains ; genetics ; Myosin Type II ; genetics ; Pedigree ; Polymerase Chain Reaction