Objective To study the effect of siRNA targeting survivin and tumor necrosis factor related apoptosis-inducing ligand on the proliferation and apoptosis of hepatocellular carcinoma ( HCC) cells. Methods siRNA eukaryotic expression vector was constructed and the effects of soluble tumor necrosis factor related apoptosis-inducing ligand ( sTRAIL) on HCC cells were observed. Results The recombinant plasmid Psilence ( + )-survivin was successfully constructed. Survivin mRNA expression inhibition ratio reached 73% by RT-PCR. Observed by MTT method sTRAIL failed to inhibit the survival rate of HepG2、HepG2/Silence( - ) cells at 12 h、24 h、48 h when compared to control groups. With the survivin gene being inhibited, the survival rate of HepG2/Silence( + ) cells(0. 518?0. 017) decreased in 12 h compared to control groups (0. 741?0. 005 ) and reached the lowest level in 48 h ( P

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

With patients' general situation, medication use, occurrence time of adverse drug reaction/event (ADR/ADE), clinical manifestations and prognosis as reference items, a retrospective study was made for 315 cases with ADR/ADE induced by Gastrodin in Chongqing from January 2008 to June 2014, in order to analyze the characteristics of ADR/ADE and provide reference for rational clinical medication. The results showed that among the 315 cases with ADR/ADE, 143 cases (45.4%) were males and 172 cases (54.6%) were females, most of them (74.9%) were aged above 45; 60 cases (19.0%) with ADE were caused by off-label indications and 66 cases (21.0%) with ADE were caused by over dosage; ADR/ADE cases induced by intravenous drip mainly happened within 30 min (85.5%), ADR/ADE cases induced by oral administration mainly happened within 2 h (74.4%), and all of ADR/ ADE cases induced by intramuscular injection happened within 10 min. Totally 593 ADR/ADE cases were reported, which were mainly damages in gastrointestinal system, skin and its adnexa; And 61.9% of ADR/ADE cases were newly reported. It is suggested that medical workers shall learn about the regularity and characteristics of ADR/ADE induced by gastrodin, apply it in clinic with standards, pay close attention to changes of patients' situations and attach importance to the monitoring of ADR/ADE, so as to enhance the safety of medication.
Adolescent ; Adult ; Aged ; Benzyl Alcohols ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; China ; epidemiology ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Gastrointestinal Tract ; drug effects ; Glucosides ; administration & dosage ; adverse effects ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Skin ; drug effects ; Young Adult

Red in vivo recombination is a new kind of genetic engineering technique based on homologous recombination. In this work, plasmid pKD46 which expresses Red recombination proteins is transferred into Escherichia coli strain DH5?.The kanamycine resistant gene is generated by PCR by using primers with homology to hisDCB gene of E.coli chromosome. Thus, the hisDCB gene was replaced with kanamycine resistant gene by the plasmid recombination system, then the resistant gene was eliminated by a helper plasmid encoding the FLP recombinase. At last, a E.coli histidine auxotroph which is sensitive to kanamycine was got. The results indicate that Red in vivo recombination is a convenient method to construct auxotrophs.

Objective To investigate the correlation of peripheral blood neutrophil-to-lymphocyte ratio(NLR)and standardized serum uric acid(SUA),i.e.,the ratio of SUA to serum creatinine(SUA/Scr),with the traditional Chinese medicine(TCM)syndrome types of early diabetic kidney disease(EDKD)from the pathogenies of deficiency-dampness-heat-stasis,thus to provide an objective basis for the precise TCM treatment of EDKD patients.Methods A retrospective study was conducted.The clinical data collection was carried out in 197 EDKD patients,including 42 cases of qi and yin deficiency syndrome(qi-yin deficiency group),44 cases of qi and yin deficiency complicated with dampness syndrome(qi-yin deficiency with dampness group),53 cases of qi and yin deficiency complicated with heat syndrome(qi-yin deficiency with heat group),and 58 cases of qi and yin deficiency complicated with stasis syndrome(qi-yin deficiency with stasis group).The differences in lipid metabolism indicators,renal function indicators,NLR,and standardized SUA of EDKD patients with various TCM syndromes were compared,and then the correlation of NLR and standardized SUA with the TCM syndromes of EDKD was investigated.Results(1)There were no statistically significant differences in the gender and glycosylated hemoglobin(HbA1c)level among EDKD patients with different TCM syndromes(P＞0.05).The age of qi-yin deficiency with stasis group was older than that of qi-yin deficiency group(P＜0.05),and the duration of EDKD in qi-yin deficiency with stasis group was longer than that in the other syndrome groups(P＜0.01).The body mass index(BMI)presented an ascending order in the qi-yin deficiency group,qi-yin deficiency with stasis group,qi-yin deficiency with heat group,and qi-yin deficiency with dampness group(P＜0.05).(2)The total cholesterol(TC)level in qi-yin deficiency group was lower than that in qi-yin deficiency with dampness group,and qi-yin deficiency with stasis group(P＜0.05),the triglyceride(TG)level in qi-yin deficiency group was lower than that in qi-yin deficiency with stasis group(P＜0.05),Scr level in qi-yin deficiency with heat group was higher than that in qi-yin deficiency group and qi-yin deficiency with dampness group(P＜0.05),the estimated glomerular filtration rate(eGFR)level in qi-yin deficiency group was higher than that in qi-yin deficiency with heat group and qi-yin deficiency with stasis group(P＜0.05),the NLR level in qi-yin deficiency with heat group was higher than that in the other three syndrome groups(P＜0.05),and the level of standardized SUA in qi-yin deficiency with stasis group was lower than that in qi-yin deficiency with dampness group and qi-yin deficiency with heat group(P＜0.05).(3)The results of the non-ordinal multinomial logistic regression analysis showed that BMI and SUA were the influencing factors for qi-yin deficiency with dampness syndrome;BMI,SUA,and NLR were the influencing factors for qi-yin deficiency with heat syndrome;BMI,TC,and standardized SUA were the influencing factors for qi-yin deficiency with stasis syndrome.The differences were all statistically significant(P＜0.05 or P＜0.01).Conclusion NLR and standardized SUA can be used as the reference indicators for the differentiation of TCM syndromes of EDKD.The elevated level of NLR may accelerate the development of qi-yin deficiency with heat syndrome,and the decreased level of standardized SUA may accelerate the development of qi-yin deficiency with stasis syndrome.

Objective To discuss the effect of psychological nursing on critically ill patients in department of thoracic surgery.Methods Totals of 56 critically ill patients after thoracic surgery were randomly divided into theexperimental group (29 cases) and the control group (27 cases).The control group received conventional nursing care while the experimental group received psychological nursing in addition.SF-36 was used before and after nursing in both groups,and incidence rates of complications and medicaldisputes were investigated and compared.Results There was no statistically significant difference in SF-60 score between two groups before nursing.After nursing,the score of role physical,physical function,body pain,vitality,social function,role emotional,psychological health and total health in SF-36 scale was respectively (87.3 ± 1.2),(84.8 ±1.0),(85.2 ±0.7),(83.2 ±0.6),(85.1 ±1.2),(74.8 ±1.0),(86.4 ±0.5) and (79.2 ±l.2)in the experimental group,higher than (86.1 ± 1.1),(78.9 ± 1.2),(74.2 ± 1.1),(73.4 ± 0.9),(79.4 ± 1.4),(65.9 ± 1.4),(77.3 ± 0.9) and(73.6 ± 1.3)in the control group,and the differences were statistically significant (t =21.051,28.903,37.812,20.962,30.873,26.761,31.080,19.050,respectively; P ＜ 0.05).The incidence rate of complications including postoperative blooding,infection and heart disorder,and medical disputes was respectively 3.4％,6.9％,6.9％ and 3.4％ in the experimental group,lower than 7.4％,11.1％,14.8％ and 29.6％ in the control group,and the differences were statistically significant (x2 =2.10,2.76,3.42,4.78,respectively; P ＜ 0.05).Conclusions Psychological nursing should fit the needs of patients' conditions,and its application can reduce the incidence of complications and disputes,so as to guarantee patients' nursing quality.

OBJECTIVETo observe the expression change of signal regulatory protein alpha1 (SIRPalpha1) in autoimmune hepatitis (AIH) and approach the relationship between SIRPalpha1 and the extent of inflammation.

METHODSImmunohistochemistry is used to detect the expression of SIRPalpha1 in the paraffin section preparations of 33 AIH and 10 normal hepatic tissue.

RESULTSSIRPalpha1 is positive or weakly positive expressed in AIH. The staining is localized in the cytoplasm of Kupffer cells in the hepatic sinusoid with focal distribution. It is negative in normal hepatic tissue. In light AIH, it is negative or weakly positive expressed with a 36.4 percent of the positive rate (4/11). The positive or strong positive expression is found in the moderate AIH with an 84.2 percent of the positive rate(16/19). There is statistical significance between both light AIH, moderate AIH and severe AIH (P less than 0.001) and moderate AIH and light AIH (P less than 0.001). There is no statistical significance between both light AIH and severe AIH (P = 0.145 ) and moderate AIH and severe AIH (P = 0.084).

CONCLUSIONSAs a negative regulatory factor, the expression of SIRPalpha1 in hepatic sinusoid Kupffer cells is some associated with the extent of AIH.

Adolescent ; Adult ; Aged ; Antigens, Differentiation ; metabolism ; Cell Communication ; Child ; Female ; Hepatitis, Autoimmune ; metabolism ; pathology ; Hepatocytes ; metabolism ; pathology ; Humans ; Kupffer Cells ; metabolism ; pathology ; Male ; Middle Aged ; Receptors, Immunologic ; metabolism ; Young Adult

PURPOSE: The purpose of this study was to investigate the influence of chronic virus-related liver disease severity on propofol requirements. MATERIALS AND METHODS: In this study, 48 male patients with chronic hepatitis B infection were divided into three groups according to Child-Turcotte-Pugh classification of liver function (groups A, B, and C with mild, moderate and severe liver disease, respectively). After intubation, propofol concentration was adjusted by +/-0.3 microg/mL increments to maintain bispectral index in the range of 40-60. Target propofol concentrations at anesthesia initiation, pre-intubation and pre-incision were recorded. RESULTS: The initial concentration used in group C was significantly lower than that used in group A or B (p<0.05), whereas no difference was observed between groups A and B. At pre-intubation, the actual required concentration of propofol increased significantly (3.2 microg/mL) in group A (p<0.05), which lead to significant differences between the groups (p<0.05). At pre-incision, the requirements for propofol decreased significantly in both groups A and B (3.0 microg/mL and 2.7 microg/mL, respectively) compared with those at pre-intubation (p<0.05), and were significantly different for all three groups (p<0.05), with group C demonstrating the lowest requirement (2.2 microg/mL). The required concentrations of propofol at pre-incision were similar to those at induction. CONCLUSION: In this study, propofol requirements administered by target-controlled infusion to maintain similar depths of hypnosis were shown to depend on the severity of chronic virus-related liver dysfunction. In other words, patients with the most severe liver dysfunction required the least amount of propofol.
Adult ; Anesthesia ; Anesthetics, Intravenous/*administration & dosage ; Chronic Disease ; Electroencephalography ; Hepatitis B, Chronic/complications/drug therapy/*surgery ; Humans ; Liver Diseases/*complications/surgery ; Male ; Middle Aged ; Piperidines/*administration & dosage ; Propofol/*administration & dosage ; Virus Diseases/*complications/surgery

OBJECTIVETo compare the effects of mannitol and hypertonic saline (HS) in treatment of intracranial hypertension (ICH) of rabbits.

METHODSThe animal mode of ICH was established by perfusing artificial cerebrospinal fluids (aCSF) with controlled pressure into the cerebral ventricles of rabbits. The mean arterial pressure, respiratory rate, tidal volume, perfusion rate of aCSF and water content of cerebrum were investigated in rabbits with ICH after a single bolus of 20% mannitol (5 ml/kg), 7.5% HS (2.2 ml/kg) or 23.4% HS (2.2 ml/kg).

RESULTSAfter the intracranial pressure was elevated from 15 cmH₂O to 75 cmH₂O, the mean arterial pressure was increased and the tidal volume was decreased. After treatment by 20% mannitol, 7.5% HS or 23.4% HS, the increased percentage of mean arterial pressure and the decreased percentage of tidal volume were similar to the changes in control group. However, the perfusion rate of CSF was increased and water content of cerebrum was decreased after treatment by either 20% mannitol or 23.4% HS, but not by 7.5% HS. No different effects were found between 20% mannitol and 23.4% HS.

CONCLUSIONWith the similar osmotic burden, 20% mannitol is more effective in treating ICH than 7.5% HS. With higher osmotic load, the efficacy of HS is enhanced, and 23.4% HS may be used as an alternative to mannitol in treatment of ICH.

Animals ; Disease Models, Animal ; Female ; Intracranial Hypertension ; drug therapy ; Male ; Mannitol ; administration & dosage ; therapeutic use ; Rabbits ; Saline Solution, Hypertonic ; administration & dosage ; therapeutic use

OBJECTIVETo investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells.

METHODSCisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay.

RESULTSMarked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 microg/mL cisplatin. Strong activation of ERK was led to by 15 microg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 micromol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 micromol/L PD 98059 at 15 and 20 microg/mL cisplatin (P < 0.05).

CONCLUSIONSCisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK activity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.

Adenocarcinoma ; enzymology ; pathology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Enzyme Activation ; drug effects ; Female ; Flavonoids ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Ovarian Neoplasms ; enzymology ; pathology ; Signal Transduction