Objective To summarize the clinical experience of serious hyponatremia resulted from applying pitnitrin to massive hemorrhage of gastrointestinal tract patients.Methods we used pitritrion on 2460 such patients and examined some symptoms of the digestive tract and nerve system,monitoring the dynamic state of blood sodium.Results The re- suits reveal that patients in 30 cases turned out to suffer from serious hyponatremia,whese sodium in the serum appeared less than 110 mmol/L,which was 1.22% of the rest subjects'.The relevant symptoms included nausea,vomit,spoor,coma and twitch etc.Following remedies included liquid supply with a high-concentrated sodium,ceasing using pituitrin,in- take of salute foods,and dynamic monitoring on the state of sodium in the serum.All patients recovered in 1 to 3 days without a single failure.Conclusion Thus we reach this conclusion:When applying pituitrin to massive hemorrhage of gastrointestinal tract patients,the above mentioned symptoms should be closely examined,and blood sodium should be timely monitored so that proper treatment should arrive in time.

Objective To evaluate the treatment of hypervascular hepatic metastasis with TACE. Methods One hundred and twenty nine cases of hepatic metastasis treated by TACE were selected retrospectively and then analyzed the survival rate,clinical effectiveness and lipidol deposition quantity in tumor.Results Malformation of tumor vessels and rich blood supply were found in all cases of this study.The survival rates of 6 months,1 year and 3 years were 100%,73.6% and 26.4% respectively.The clinical effective rate was 68.2%(88/129)and no-progress rate was 23.3%(30/129).The satisfactory lipidol deposition quantity was obtained in 80.9%(97/129).Conclusions TACE is a favorable method for hepatic metastasis,and discerning the hypervascular subgroup could improve the treating effectiveness and be useful to make an appropriate planning.

Aim To investigate the role of P2X7 recep-tor and its mediated NLRP3 inflammatory signaling pathway in alcohol-induced liver injury. Methods The acute alcoholic liver injury model was established by NIAAA method, and thirty C57BL/6 male mice were randomly divided into three groups (n =10):control group, model group, A438079 group, The three groups were processed as follows in the last week:control group and model group: given an equal dose of saline intraperitoneal injection(about 0.2 mL/only) once a day. According to the weight of the mice, A438079 group was given intraperitoneally injection by 200 μmol·kg-1of A-438079 (prepared at 7 g·L-1 of A438079,about 0.2 mL/only) once a day. And it was given a single 31.5% alcohol solution by intragas-tric administration on the last day of the morning,with the dose of 10 mL·kg-1. Nine hours later alanine aminotransferase (ALT), aspartate aminotransferase (AST),cholesterol(TCHO),triglyceride(TG) were measured by orbital blood in mice. HE staining was used to observe the pathological changes of the liver. Immunohistochemical method was applied to detect the expression of P2X7R in liver tissues. Western blot was employed to detect the levels of P2X7R, NLRP3, ASC, IL-1β and IL-18 in liver tissues. Results Compared with control group,the levels of ALT,AST, TG and TCHO in model group were significantly en-hanced, and the liver injury was obvious. Compared with model group, the levels of ALT, AST, TG and TCHO in A438079 group significantly decreased. Compared with control group, the expressions of P2X7, NLRP3, ASC, IL-1β, IL-18 in model group were significantly higher than those in control group. Compared with model group, the expression levels of P2X7, NLRP3, ASC, IL-1β and IL-18 in A438079 group significantly decreased. Conclusion Alcohol-induced liver injury may be associated with P2X7R-NLRP3 signaling pathway.

Saliva and blood were collected from two patients who had not received post exposure prophylaxis in the cities of Wenzhou and Xinning respectively. Both patients were confirmed as positive for rabies by detection of rabies virus specific nucleoprotein antibodies in the sera by Western Blot. However, rabies virus specific RNA was only identified in the saliva collected from the patient in Wenzhou. Furthermore, the isolate Zhejiang Wz0 (H) was obtained by inoculating one-day-old suckling mice. Both nucleoprotein (N) and glycoprotein (G) genes from the isolate were amplified by RT-PCR and sequenced. Phylogenetic analysis indicated that the isolate belonged to classic rabies virus, and shared a higher homology with the street viruses from dogs in the main endemic areas in China and the street virus from dogs in Indonesia than with other known strains. Further comparison of the deduced amino acid sequences between the isolate and the vaccine strains used in China showed that the virus had a higher level of homology with the vaccine strain CTN than with the other vaccine strains (3aG, PV, PM and ERA). In particular, amino acid residues substitutions located in antigenic site Ⅲ in the G protein, which could react with the neutralizing antibodies, were observed. These results suggested that the virus belonged to the classic rabies virus, and both N and G genes diverged from the current vaccine strains used in China at either the nucleotide or the amino acid level.

OBJECTIVEIn order to obtain functional genes, a normalized stems cDNA library was constructed from medicinal plant Dendrobium officinale.

METHODSMART (switching mechanism at 5' end of RNA transcript) cDNA synthesis combined with DSN (duplex-specific nuclease) normalization was applied to construct the normalized full-length cDNA library of D. officinale.

RESULTThe titer of cDNA library was about 1.3 x 10(6) cfu x mL(-1) and the average insertion size was about 1.5 kb with high recombination rate (93.9%). Random selected 163 positive clones were sequenced at single side. Bio-information analysis indicated that 147 from 150 high-quality unique sequences matched corresponding homologous proteins, and they participated in various biological processes based on GO (gene ontology). There were 8 clones with complete coding sequence, which presumed to be full-length genes.

CONCLUSIONThese results showed preliminarily that we successfully constructed a normalized full-length cDNA library of D. officinale which could be used to screen the functional genes related to metabolic pathways of medicinal ingredients.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; biosynthesis ; genetics ; Dendrobium ; genetics ; Gene Library ; Molecular Sequence Data ; Plants, Medicinal ; genetics ; RNA, Messenger ; genetics ; metabolism ; Sequence Analysis, DNA ; methods

The cytoplasm of E. coli is a reducing environment where cysteines do not engage in disulfide bonds. Any disulfide bonds that do appear are rapidly reduced through the action of disulfide reducing enzymes such as thioredoxin and glutaredoxin. To study the influence of E. coli cytoplasm on the solubility of recombinant proteins produced in it, bovine fibroblast growth factor (BbFGF), with single disulfide bond, and anti-HBsAg single-chain Fv (HBscFv), with two disulfide bonds, were selected as the pattern molecules of simple protein and complex protein, respectively. pJN98-BbFGF, a BbFGF expressing plasmid based on the vector pET3c, was constructed and transformed into normal host BL21(DE3) and a reductase deficient strain, E. coli Origami(DE3). At the same time, pQE-HBscFv, a HBscFv expressing plasmid was constructed and transformed into M15 [pREP4] and Origami(DE3). The recombinant BbFGF and HBscFv were produced in 2 types of bacteria and their solubilities and bioactivities were determined, respectively. It was found that the majority of BbFGF had formed inclusion body in the cytoplasm of BL21 (DE3) and all of them turned into soluble protein in Origami(DE3). It was also found the productivity of BbFGF in Origami (DE3) was 5% - 10% of the total protein and the value was 15% - 23% in BL21(DE3). BbFGFs produced in 2 recombinant bacteria were purified by cation exchange and heparin affinity chromatography. MTT assay revealed that the bioactivity of BbFGF purified from Origami(DE3) was higher than its counterpart from BL21(DE3). The ED50 of BbFGFs from different bacteria was 1.6ng/mL and 2.2ng/mL, respectively. As far as HBscFvs, both of them formed inclusion body in the cytoplasm of M15 [pQE-HBscFv] and Origami [pQE-HBscFv]. The inclusion body was solubilized in 6mol/L GuHCl, purified with a His-Trap column and then refolded by dialysis step-by-step against buffers containing downtrend concentration of GuHCl. Indirect ELISA was applied to determine the HBsAg binding activity of HBscFvs. It was found there was no obvious difference between the bioactivity of refolded HBscFvs produced from 2 recombinant bacteria. On the other hand, the supernatant of Origami [pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15 [pQE-HBscFv] displayed without any bioactivity. The soluble HBsFv in the cytoplasm of Origami [pQE-HBscFv] was purified by cation exchange and immobilized metal affinity chromatography (IMAC) and the yield was 1 - 2mg/L. Those results suggested that modification of the redox environment of E. coli cytoplasm greatly improved the solubility of recombinant disulfide-bonded proteins produced in it. In the next step, we had like to co-express of molecular chaperones or refoldase to raise the yield of soluble recombinant proteins, as well as optimizing the culture condition of the "oxidizing" E. coli.
Animals ; Antibodies ; genetics ; immunology ; metabolism ; Cattle ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; enzymology ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fibroblast Growth Factors ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hepatitis B Surface Antigens ; immunology ; Inclusion Bodies ; chemistry ; metabolism ; Oxidoreductases ; genetics ; Plasmids ; genetics ; Protein Engineering ; Recombinant Proteins ; chemistry ; genetics ; metabolism ; Solubility

Stroke research and rehabilitation have traditionally focused on the physical and functional impact of a stroke. Less attention has been given to the psychosocial factors associated with this chronic condition. By the few studies that have specifically focused on psychosocial factors in the context of stroke, poststroke depression is demonstrated to significantly influence stroke outcomes. Associations of stroke with psychological symptoms other than depression have rarely been evaluated. This study was aimed to investigate the changes of psychological stress, social support and medication adherence in patients with ischemic stroke in the mainland of China. In this study, 90 patients with hemiplegia one year after first-ever middle cerebral artery infarction (stroke group) in the Zhongnan Hospital of Wuhan University from June 2008 to June 2011 were recruited for interview. Ninety age- and sex-matched normal volunteers (control group) were also examined at the same period. The psychological distress was assessed by the Symptom Checklist 90 (SCL-90), the social support by the Social Support Rating Scale (SSRS), and medication adherence by Morisky's self-reported inventory, respectively. Group differences were analyzed using unpaired-t test and chi-squared test. The results showed that total mean scores of the SCL-90 in the stroke group were higher than those in the control group (P<0.01). Except two dimensions, paranoid ideation and psychoticism, mean scores of the rest dimensions (including somatization, obsession-compulsion, interpersonal sensitivity, depression, anxiety, hostility, and phobic anxiety) of SCL-90 were significantly higher in the stroke group than those in the control group (P<0.05, or P<0.01). The objective support, subjective support, support availability and total social support scores in the stroke group were significantly higher than those in the control group (P<0.05, or P<0.01). Those in the "SCL-90 total scores >150 group" were significantly higher than in the "SCL-90 total scores <100 group" and the "SCL-90 total scores between 100 to 150 group" (P<0.05, or P<0.01). Those in the "SCL-90 total scores between 100 to 150 group" were significantly higher than in the "SCL-90 total scores <100 group" (P<0.05). In 90 patients with ischemic stroke, 26 (28.89%) patients obtained high medication adherence, 47 (52.22%) patients medium medication adherence, and 17 (18.89%) patients low medication adherence, respectively. Among these stroke patients, there were 17 (50.00%) patients with high medication adherence in the "SCL-90 total scores >150 group", 28 (75.67%) patients with medium medication adherence in the "SCL-90 total scores between 100 to 150 group", and 12 (61.16%) patients with low medication adherence in the "SCL-90 total scores <100 group", respectively. There was significant difference in the medication adherence rate among the different SCL-90 scores groups in these stroke patients (P<0.05 or P<0.01). It was led to conclude that ischemic stroke patients one year after hemiplegia have psychological distress, low level of social support and poor medication adherence in the mainland of China. Therefore, it is necessary to mobilize the government, medical institutions and various social support groups to offer psychological interventions to relieve the stress of patients with ischemic stroke, and improve their medication adherence.
Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; psychology ; China ; Female ; Humans ; Infarction, Middle Cerebral Artery ; drug therapy ; psychology ; Male ; Medication Adherence ; statistics & numerical data ; Middle Aged ; Social Support ; Stress, Psychological ; etiology ; psychology ; Surveys and Questionnaires

The introduction of artificial intelligence into the medical field is also facing social ethical challenges while promoting changes in the field of health services. To realize the healthy development of medical artificial intelligence, we must consider the ethical issues inherent in the application of artificial intelligence technology in medical services. The article analyzes the ethical issues brought about by medical artificial intelligence from the aspects of security, doctor’s principal status, doctor-patient relationship, and fair benefit. By implementing special supervision, clarifying the principal status of doctors, and strengthening ethical protocols, the application of artificial intelligence in medical services will be guided to promote artificial intelligence to better improve the service capabilities and quality of healthcare system.

OBJECTIVE: To construct the recombinant lentivirus RNAi vector, and to determine whether the lentivirus mediated short hairpin RNA (shRNA) can inhibit the tissue factor (TF) expression in endothelial cells. METHODS: Two short hairpin RNAs targeting to human TF were cloned into pENTRTM/U6 plasmid to obtain an entry clone, and the positive clones were verified by sequencing. A recombination reaction was performed between the pENTR/U6 entry construction and pLenti6/BLOCKiTTM-DEST vector, and then the positive clones were confirmed by sequencing. The 293FT cell line was transfected by the above recombined plasmid and lentivirus packing materials, the culture supernatant was harvested, and the virus titer was determined. RT-PCR and ELISA were used to observe the inhibition of TF gene expression after the lentivirus transduction in human umbilical vein endothelial cells. RESULTS: The shRNA sequences targeting to human TF were cloned into the vectors, and an entry clone and an expression clone were constructed successfully, which were proved by sequence determination. Viral particles were packaged in the 293FT cell line, all virus stocks were collected, and the transfection titer was 5*10(5)/transduced unit. RT-PCR and enzyme linked immunosorbent assay demonstrated that the lentivirus stocks could suppress the TF expression in endothelial cells remarkably. CONCLUSION Lentivirus RNAi vectors containing human TF gene are successfully constructed, and lentivirus mediated shRNA can inhibit the TF expression in endothelial cells, which may provide a highly effective method for the prevention and treatment of thrombo-embolic diseases.
Base Sequence ; Down-Regulation ; Endothelial Cells ; cytology ; metabolism ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; Molecular Sequence Data ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Thromboplastin ; biosynthesis ; genetics ; Umbilical Veins ; cytology

This paper described a rapid and sensitive HPLC method to analyze (E)-3,5,4'-trimethoxystilbene (BTM-0512) in rat plasma and tissues. The analysis used a BDS Hypersil C18 analytical column (250 mm x 4.6 mm ID, 5 microm) and acetonitrile/water as the mobile phase. The UV detection wavelength was 319 nm. Proteins were precipitated with acetonitrile and diethylstilbestrol as internal standard. The method was validated according to State Food and Drug Administration of China and ICH of Technical Requirements for Registration of Pharmaceuticals for Human Use Guidelines. The limit of detection (S/N: 3/1) for BTM-0512 was 0.005 microg x mL(-1) for plasma. The method performances were shown to be selective for BTM-0512 and the linearity of the assay method was up to 10.0 microg x mL(-1) and 40.0 microg x g(-1) for plasma and tissues, respectively. At 0.1, 1 and 5 microg x mL(-1) (n=5), intraday and interday precision values (% RSD) were in the range of 2.6% - 5.1% and 2.4% - 4.8%, respectively. Mean accuracy and absolute recoveries of BTM-0512 ranged from 95.3% - 100.1% and 95.9% - 100.9% for plasma and tissues, respectively. This method can be quite useful for BTM-0512 pharmacokinetic and tissue distribution studies, for purpose which multiple plasma and tissue samples can be analyzed quickly with high reproducibility.
Angiogenesis Inhibitors ; blood ; pharmacokinetics ; Animals ; Anti-Allergic Agents ; blood ; pharmacokinetics ; Antineoplastic Agents ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Spectrophotometry, Ultraviolet ; methods ; Stilbenes ; blood ; pharmacokinetics ; Tissue Distribution