Objective To evaluate the sensitivity of three different clinical consciousness scales to patients with disorders of consciousness.Methods A total of 84 patients in vegetative state (VS) were selected and scored by the consciousness scales including Glasgow Coma Scale (GCS),Coma Recovery Scale-Revised (CRS-R) and Chinese Vegetative State Scale (CVSS) respectively.The patients were followed every 2 weeks for 12 weeks or every 4 weeks for 12-24 weeks.The assessment finished in 6 months or till the time when patients emerged from minimally conscious state ( MCS.The diagnostic sensitivity and the sensitivities of each sub-scale in CRS-R were evaluated when the patients with MCS emerged from VS.The diagnostic consistency with other scales was observed and analyzed by Spearman correlation as well.The Spearman -relate was used to analyze the correlations among the total scores of three scales in the patients on admission (36 patients with acute and 48 patients with chronic phase).Results Of 84 VS patients,there were 47 emerged to MCS.CRS-R was the most sensitive in the diagnosis of MCS (P ＜ 0.05 ) among all scales.The visual subscale is the most sensitive among all the CRS-R subscales ( P ＜0.01 ).Among the 84 patients,the remaining 37 were diagnosed as VS by all three scales.The total scores correlation analysis showed that the score of CRS-R significantly correlate with that of GCS in acute and chronic phase ( P ＜0.01 ) and significantly correlate with that of CVSS only in the acute stage ( P ＜ 0.05 ).Conclusions It is indicated that CRS-R,especially the visual subscale of CRS-R is the more sensitive tool for detecting.MCS during recovery process in the disorders of consciousness.As to the patients with deep unconsciousness and with no significant recovery,the three instruments are in good agreement to determine the degree of disorders of consciousness.It is suggested that CRS-R can be widely used to assess the level of consciousness in various stages.

Animals ; Brain Diseases ; parasitology ; Child ; Female ; Humans ; Sparganosis ; Sparganum

Objective To summarize the experience of treating the end stage of liver disease complicated with renal failure using combined liver-kidney transplantation.Methods The clinical data of 28 cases receiving combined liver-kidney transplantation were retrospectively analyzed, including the inclusion criteria of surgical indications, modus operandi, protocol of immunosuppression and the prognosis post-operation.Results Among these 28 cases in our study, 22 cases suffered from liver and renal failure, accounting for 78.6%; 4 cases were diagnosed as having hepatorenal syndrome, accounting for 14.3%; and 1 case had hyperoxaluria and polycystic liver with polycystic kidney. As for the modus operandi we used, piggy-back procedure was adopted for 4 patients and classic procedure without bypass was used for the rest. Donor kidneys were all put in the right iliac fossa. During the follow-up period of 5 months to 7 years, one-and 3-year survival rate of the recipients was 92.9% and 78.3% respectively. Among these 28 recipients, 4 cases had the graft renal dysfunction early post-operation: One died and 3 recovered through consecutive therapy. One case received re-transplantation of the liver 3 months after the first due to the relevant complications and then recovered. During this period, no impact on the renal function occurred. Eleven cases had pulmonary infection post-operation, and 1 died. No acute rejection occurred.Conclusion Combined liver-kidney transplantation is the effective treatment to the patients with end stage liver disease complicated with renal dysfunction. Suitable case selection and perfect operation timing were the key points to the success of combined liver-kidney transplantation.

AIM:To investigate the effect and mechanism of osthole on increasing the cytotoxicity of doxorubi -cin(DOX)to prostate cancer cells.METHODS:MTT assay was performed to evaluate the viability of LNCaP cells trea-ted with osthole and DOX.The protein expression of silent information regulator 1(SIRT1),p53,acetylated p53 and Pu-ma,as well as release of cytochrome C and activation of caspase-9 and caspase-3 in the LNCaP cells treated with osthole and DOX were determined by Western blot.The apoptosis of the LNCaP cells treated with osthole and DOX was analyzed by flow cytometry.RESULTS:Osthole significantly increased the cytotoxicity of DOX against p 53-wildtype prostate cancer cell line LNCaP.Osthole significantly inhibited the expression of SIRT 1 in the LNCaP cells.Transfection with SIRT1 plas-mid decreased the cytotoxicity of osthole and DOX co-treatment against LNCaP cells.Combination with osthole and DOX significantly induced the over-expression and acetylation of p53.Transfection with p53 siRNA significantly decreased the synergistic effect of osthole on cytotoxicity of DOX-treated LNCaP cells.Combination with osthole and DOX significantly in-duced the release of cytochrome C into the cytoplasm from mitochondria,followed by activation of caspase-9 and its down-stream molecule caspase-3,thus leading to cell apoptosis in the LNCaP cells.CONCLUSION:Osthole promotes the p53-dependent apoptosis in DOX-treated prostate cancer LNCaP cells by down-regulating the expression of SIRT1.

OBJECTIVETo study the genotype of swines isolated from humans and their relationships with hepatitis E virus (HEV) in the rural areas of southern China.

METHODSSpecimens collected from normal people with HEV-IgM positive, acute hepatitis E patients and from swine in the same area were detected for HEV RNA using RT-nPCR with ORF2 primers. The positive PCR products were cloned and sequenced.

RESULTS13 out of the 132 samples from swine stool, 4 of 26 HEV-IgM positive sera of normal people and 1 of 4 acute hepatitis E patients' stool sample and sera were tested positive for HEV RNA. Data from sequence analysis showed that the identity at nucleotide level was 89.3%-100.0% among the 10 isolates which shared 78.7% - 84.7%, 83.3% - 85.3%, 76.0% - 80.0% and 84.7% - 95.3% nucleotide sequence identity with HEV genotype I, II, III and IV respectively in the region (nt6317- 6466).

CONCLUSIONHEV circulating in humans and swine in the area belonged to genotype IV.

Animals ; China ; Cloning, Molecular ; Feces ; virology ; Genes, Viral ; Genotype ; Hepatitis E ; veterinary ; virology ; Hepatitis E virus ; genetics ; isolation & purification ; Humans ; Immunoglobulin M ; blood ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Rural Health ; Swine ; Swine Diseases ; virology

AIMTo develop a HPLC-ESI-MS assay for determination of eperisone hydrochloride in human plasma and investigate the pharmacokinetics and bioequivalence of two eperisone hydrochloride tablets in human.

METHODSBuflomedil hydrochloride was used as the internal standard. After alkalized with saturated sodium bicarbonate solution, plasma was extracted with diethylether-cyclohexane (1:1) and separated using HPLC on a reversed-phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate buffer solution (adjusted to pH 3.88 with acetic acid)-methanol (20:80). HPLC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 260 for eperisone and m/z 308 for the internal standard. A randomized crossover design was performed in 20 healthy volunteers. In the two study periods, a single 100 mg dose of each tablet was administered to each volunteer.

RESULTSCalibration curve was linear over the range of 0.02-20 microg x L(-1). The limit of quantification for eperisone hydrochloride in plasma was 0.02 microg x L(-1). The main pharmacokinetics parameters T1/2, Tmax and Cmax were (2.7 +/- 0.4) h, (1.1 +/- 0.5) h and (2.8 +/- 2.8) microg x L(-1) for the reference tablet; (2.8 +/- 0.5) h, (1.1 +/- 0.4) h and (3 +/- 4) microg x L(-1) for the test tablet, respectively. The relative bioavalability of the test tablet was (101 +/- 13)%.

CONCLUSIONThe assay was proved to be sensitive, accurate and convenient. The two formulations were bioequivalent.

Adult ; Chromatography, High Pressure Liquid ; Humans ; Male ; Propiophenones ; administration & dosage ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Therapeutic Equivalency

Objective To compare the efficacy of ultrasound-guided fine-needle aspiration(US-FNA)biopsy in diagnosing solid and complex thyroid nodules with different size. Methods One hundred and seventy-five thyroid FNA biopsies were prospectively performed on 168 patients ranging from 4 to 75 years of age. Sixty-three nodules were surgically excised and the others were clinically followed-up. The cytology diagnoses were categorized into four groups: benign, malignant, suspicious and unsatisfactory. Results There was no significant complication in the all 115 solid and 60 complex thyroid lesions and there were 36and 3 malignant nodules respectively in solid and complex thyroid nodules. The nondiagnostic rates of solid and complex nodules were 7％ and 8％. The accuracy of US-FNA in diagnosing complex thyroid nodules was comparatively equal to that of in solid thyroid nodules. In solid thyroid nodules, the sensitivity and accuracy in ≤1 cm group were similar to that of in ＞1 cm group. Conclusions US-FNA was an accurate and reliable method to diagnose thyroid solid and complex lesions.

OBJECTIVETo investigate angiogenesis of collagen-chitosan porous scaffold, and to study survive of skin grafts on the scaffold after bilayer dermal equivalent (BDE) was transplanted on wounds with full thickness skin defects.

METHODSThe full thickness skin defects were made on 10 Bama miniature pigs and the BDE composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wound. Angiogenesis in dermal equivalent, wound healing, and healing and survive of skin grafts on dermal equivalent were observed in 1, 2, and 3 weeks after the BDE transplantation. At the same time, CD34 positive signals (neo-forming micro-vessels) were detected by immunohistochemical staining.

RESULTSInflammatory cells and fibroblasts infiltrated into dermal equivalent and a few new micro-vessels had been formed in 1 week after the BDE transplantation; neo-forming micro-vessels perpendicular to wound bed had increased significantly in 2 weeks after the BDE transplantation; neo-forming micro-vessels could be observed in almost all dermal equivalents in 3 weeks after the BDE transplantation. CD34 positive signals (neo-forming micro-vessels) in 3 weeks after the BDE transplantation was much more than those in 2 weeks after the BDE transplantation, and CD34 positive signals in 2 weeks after the BDE transplantation was much more than those in 1 week after the BDE transplantation. Survival rate of intermediate split thickness skin graft were 10%, 70% and 100% respectively after the skin grafts had been grafted for 2 weeks on surface of the scaffold which had been transplanted for 1, 2 and 3 weeks. Epidermis which had been grafted on surface of the scaffold for 1 or 2 weeks could perfectly survive after BDE had been transplanted for 1 or 2 weeks.

CONCLUSIONSCollagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect and can induce fibroblast infiltration and new micro-vessel formation. Epidermis grafted on surface of collagen-chitosan porous scaffold can perfectly repair wounds, and it has brilliant applied prospects in repairing skin defect.

Animals ; Chitosan ; Collagen ; Disease Models, Animal ; Female ; Graft Survival ; Neovascularization, Physiologic ; Silicones ; Skin ; injuries ; Skin Transplantation ; Swine ; Swine, Miniature ; Tissue Scaffolds ; Wound Healing

The expression plasmid pET32CPS harboring SmCPS gene was transformed into E. coli BL21 trxB (DE3) resulting in recombinant strain E. coli [pET32CPS]. The induction of E. coli [pET32CPS] in different temperatures, induction time, IPTG concentrations and A600 values of E. coli were performed. The optimal expression conditions of SmCPS were characterized according to the orthogonal analysis, and the ratio of the interest protein to total proteins reached to 35.6%. The recombinant SmCPS protein purified by Ni2+ affinity chromatography column was identified by SDS-PAGE and Western blotting, and then used for rabbit immunization. The titer of the rabbit antiserum against SmCPS was about 1:24 300 after the third immunization, and could specifically recognize the antigen of SmCPS protein by Western blotting analysis. The successful preparation of polyclonal antibody against SmCPS laid a foundation for further correlative study between expression of SmCPS and the production of tanshinones in protein level.
Alkyl and Aryl Transferases ; genetics ; isolation & purification ; metabolism ; Animals ; Antibody Formation ; Escherichia coli ; metabolism ; Gene Expression ; Immune Sera ; biosynthesis ; immunology ; Isopropyl Thiogalactoside ; chemistry ; Male ; Plant Proteins ; genetics ; isolation & purification ; metabolism ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Plasmids ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Temperature ; Time Factors ; Transformation, Genetic

Adult ; Aged ; China ; ethnology ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; genetics ; Polymorphism, Genetic ; Thrombospondin 1 ; genetics