Objective To observe the effect of Shenfu injection on fluid intake volume of resuscitation therapy for patients with septic shock. Methods The clinic data of 36 patients with septic shock admitted to Department of Critical Care Medicine of the Affiliated Hospital of Hangzhou Normal University from June 2010 to June 2013 were retrospectively analyzed. All the patients were treated with western conventional medicine. Twenty cases treated with western medicine combined with Shenfu injection (intravenous drip 100 mL once daily, half of a month was a therapeutic course) were defined as Shenfu group; the rest 16 cases treated with western medicine only were assigned as control group. The following data after treatment for 6, 24, and 72 hours in the two groups were compared:liquid intake and urine volumes, system vascular resistance index (SVRI), mean arterial pressure (MAP), cardiac index (CI), and case fatality rate in 28 days. Results There were no significant differences in the liquid intake volume in 6 hours after treatment (mL:3 101±219 vs. 3 329±295, P>0.05), the urine volumes in 6, 24 and 72 hours after treatment (mL, 6 hours:701±229 vs. 651±292, 24 hours:1 870±566 vs. 1 697±618, 72 hours:7 396±2 546 vs. 5 987±2 497), and the levels of SVRI in 24 hours after treatment between Shenfu group and control group (kPa·s·L-1·m-2:802±158 vs. 741±106, all P>0.05). The total liquid intake volumes (mL) in 24 hours and 72 hours after treatment in Shenfu group were significantly less than those in the control group (24 hours:4 544±425 vs. 4 996±396, 72 hours:10 985±891 vs. 11 612±807, both P<0.05). The SVRI, MAP, and CI in 72 hours of Shenfu group were significantly higher than those of control group [SVRI (kPa·s·L-1·m-2): 1 361±182 vs. 1 163±183, MAP (mmHg, 1 mmHg = 0.133 kPa): 76.2±6.1 vs. 71.8±6.3, CI (mL·s-1·m-2):76.2±7.5 vs. 70.8±7.2, all P<0.05], and the 28-day mortality rate in Shenfu group was significantly lower than that of control group [25.0%(5/20) vs. 62.5%(10/16), P<0.05]. Conclusion The application of Shenfu injection was favorable to the reduction of liquid intake volume in 72 hours after treatment that may be beneficial to the fluid limitation management in the course of treatment for septic shock.

OBJECTIVETo investigate the influence of the smoke tar on the expression of aromatic hydrocarbon receptor (AHR) and the cytochrome P4501Al (CYP1A1) gene of mice lungs.

METHODSThe smoke tar of 5.29, 10.58 and 15.87 mg/kg was administered intraperitoneally in mice respectively. RNA of mice lungs was got with RNA kit. RT-PCR technique was used for determining AHR and CYP1A1 gene expression with beta-actin as control.

RESULTSThe AHR gene expression level was (0.554 +/- 0.023) for the mice intraperitoneally administered with 5.29 mg/kg smoke tar for 72 hours with the significant difference in gene expression level compared with the Tween-80 group (0.484 +/- 0.045) (P < 0.05). The AHR gene expression levels were (0.555 +/- 0.014), (0.606 +/- 0.051), and (0.566 +/- 0.014), (0.684 +/- 0.069) for the mice intraperitoneally administered with 10.58 and 15.87 mg/kg smoke tar for 48 hours and 72 hours respectively with the significant difference in gene expression level compared with the Tween-80 group (0.486 +/- 0.060, 0.484 +/- 0.045) (P < 0.05, P < 0.01). The CYP1Al gene expression levels were (1.535 +/- 0.021), (1.643 +/- 0.046) and (1.624 +/- 0.056), (1.739 +/- 0.038) respectively with the significant difference compared with the Tween-80 group (l.436 +/- 0.016, 1.404 +/- 0.036) (P < 0.01).

CONCLUSIONThe smoke tar can regulate up the expression of AHR and CYP1A1 gene at a certain dosage and time. The regulation of the smoke tar for the expression of AHR was earlier than for that of CYP1A1.

Animals ; Cytochrome P-450 CYP1A1 ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Female ; Gene Expression ; drug effects ; Lung ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Aryl Hydrocarbon ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Smoking ; Tars ; toxicity ; Up-Regulation

Background Whether ocular anterior and posterior chamber exist a blood-aqueous barrier is in controversy.Conventional method can not offer a good evidence because it is unable to detect the aqueous component in the posterior chamber.Objective This study was to investigate the distribution of Gadolinium-diethylene triamine pentaacetic acids(Gd-DTPA)after peripheral iridectomy with magnetic resonance imaging(MRI)in rabbit.Methods Monocular peripheral iridectomy was performed on the right eyes in 8 clean New Zealand white rabbits and the fellow eyes were as controls.0.2 ml/kg(0.5 mol/L)Gd-DTPA,a tracer of MRI,was injected into ear vein in vivo to scan the eyes with MRI for the observation of the permeability and distribution.The signal enhanced ratio of interest region associated with time were analyzed.Results The signal in ciliary body of both eyes showed an immediately sharp enhancement within 10 minutes following the injection of Gd-DTPA with a peak intensity at 30-40 minutes,and then the intensity was gradually weaken over time.The signal was stronger in the operative eyes than that in the fellow eyes.The signal in the posterior chamber was gradually increased after operation,however,that in posterior chamber of the control eyes was lower.The interest regions of Gd-DTPA were ciliary,anterior chamber and posterior chamber,and the enhanced signal intensities were consisted in the posterior chamber after operation.However,the increase of the signal was not seen in the posterior chamber in the control eyes.Conclusions The pathway of plasma protein entering into the anterior chamber is very different from that of aqueous secretion.There exists a barrier between the anterior and posterior chamber which might be an integral part of the blood-ocular barrier.

PTEN, a dual-specificity phosphatase, exerts its tumor-suppressive effects through the inhibition of cell cycle progression and cell immigration, therefore could be an important candidate for tumor-suppression. As study on prokaryotic expression of PTEN and its anti-tumor functions has not been reported, the present study aims at an efficient expression of functional PTEN in Escherichia coli and the investigation of its tumor-suppression activity. PTEN cDNA cloned in our lab previously was recombined into prokaryotic expression vector pET-44a(+) to construct pET-PTEN (pEP) and pET-Nus-PTEN (pENP). PTEN was fused with 6 x His tag in pEP, and with Nus in pENP, which could be useful for a stable and soluble expression. The recombinant vectors were transformed into both BL21 (DE3) (BL) and Rosetta-gami (DE3) pLysS (RG). The former is a normal expression host while the latter is optimized for expression of eukaryotic genes and folding of target proteins. On the induction of 0.5mmol/L IPTG, 55kD and 118kD specific protein bands were observed, corresponding to His-PTEN and Nus-PTEN fusion proteins, respectively. Western blot analysis showed the recombinant fusion proteins could react with PTEN polyclonal antibody. The recombinant HTEN was expressed both in soluble fraction and inclusion body. Higher expression levels of recombinant PTEN were obtained in BL (His-PTEN: 10.3%; NusA-PTEN: 18.7%), whereas the higher percentages of soluble recombinant proteins were observed in RG (His-PTEN: 4.7%; Nus-PTEN: 6.6%). The obtained recombinant fusion proteins were purified by affinity chromatography and were showed to be homogeneous in SDS-PAGE. In tumor-suppression experiments, His-PTEN was proved to have significant inhibition on growth of mice solid tumor with an inhibitory ratio of 58.76%, and on the proliferation of DU-145 tumor cells with an inhibitory ratio of 46.16%. The cell cycle progression of DU-145 tumor cells was also arrested from G0/G1 to S phase. His-PTEN from RG was proved to have significantly higher tumor-suppression activity than that from BL, indicating that there may be some advantages for eukaryotic genes to be expressed in the former host. This is the report of functional recombinant PTEN expressed in Escherichia coli. Purified His-PTEN was used for immunizing Kunming mice, and ascitic polyclonal antibodies raised against His-PTEN were generated using sarcoma 180 cells. At 1:2000 dilution, the antibodies could interact with PTEN by western blot. The present study has laid a foundation for application of PTEN in cancer therapy.
Animals ; Antibodies, Neoplasm ; biosynthesis ; immunology ; Antineoplastic Agents ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Humans ; Liver Neoplasms, Experimental ; therapy ; Mice ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo study the polymorphism of CYP1A1 gene Msp I site in the Mongolian and Han nationality populations of Inner Mongolia.

METHODSThe PCR-restriction fragment length polymorphism(PCR-RFLP) technique was used to analyze the genotypes of CYP1A1 gene Msp I site in 80 subjects of Mongolian nationality and 120 subjects of Han nationality among whom there is no blood relationship each other.

RESULTSThe genotype frequency of CYP1A1 gene Msp I site showed that the wild-type, heterozygote, homozygous variants were 35.0%, 48.7%, 16.3% and 33.3%, 52.5%, 14.2% respectively distributions of Mongolian nationality and Han nationality population, and the Chi-square tests showed that there was no significant difference between the two groups.

CONCLUSIONThe genotype frequency distributions of CYP1A1 gene Msp I site did not exhibit the obvious difference between Mongolian nationality and Han nationality population of Inner Mongolia.

Adolescent ; Adult ; Aged ; Binding Sites ; genetics ; China ; Cytochrome P-450 CYP1A1 ; genetics ; Deoxyribonuclease HpaII ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Mongolia ; ethnology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length ; Young Adult

Objective To compare the differences in virulence-related factor aspartate protease,biofilm formation,and gene expression among clinical isolates of Candida parapsilosis(C.parapsilosis).Methods Gene sequencing and microsatellite typing(MT)method were adopted to identify C.parapsilosis isolated from patients with clinical fungal infection.The production of secreted aspartate protease and biofilm formation ability of each strain were de-tected,and the expression of biofilm formation related-genes BCR1,EFG1,and HWP1,as well as aspartate prote-ase virulence genes SAPP1,SAPP2,SAPP3 were compared among the strains.Results A total of 8 clinically iso-lated C.parapsilosis strains were collected,all of which were identified as genotype Ⅰ.Based on microsatellite ty-ping results,8 clinical strains were divided into 4 microsatellite types.G1,G2,and G3 strains isolated from the urine,peripherally inserted central catheters(PICC),and blood of patient A were of different subtypes.J1,J2,J3,J4,and J5 strains were of the same type,and isolated from blood specimens of patient B at different periods.All 8 clinical strains could form biofilm,and their biofilm formation ability was higher than that of the standard strain of C.parapsilosis(ATCC 22019).G1,G3 and J5 strains had strong biofilm formation ability,J1,J2,J3,and J4 strains had moderate biofilm formation ability,and G2 strain had weak biofilm formation ability.All of the eight clinical isolates secreted aspartate protease,and their in vitro expression levels of the enzyme were higher than that of the standard strain(ATCC 22019).G3,G1,and G2 strains showed low,moderate,and high in vitro enzyme expression respectively,with statistical differences(all P＜0.05).Enzyme expressed moderately in J1 and J5 strains,and highly in J2,J3,and J4 strains.Difference between moderate and high expressions was statistically significant(P＜0.05).The expression levels of biofilm formation genes BCR1,EFG1,and HWP1 in various strains isolated from patients A and B increased.In strains isolated from patient A,the expression level of EFG1 gene in G1 strain was higher than that in G2 strain(P＜0.05).There was no statistically significant difference in BCR1,EFG1,and HWP1 gene expression levels among strains isolated from patient B.The expression levels of as-partate protein genes(SAPP1,SAPP2,and SAPP3)in various strains isolated from patients A and B increased.The expression levels of SAPP1 and SAPP2 in strain G1 were higher than those in G2 and G3(both P＜0.05).There was no statistically significant difference in the expression levels of SAPP1,SAPP2,and SAPP3 genes in strains from patient B.Conclusion Clinical isolates of C.parapsilosis have higher biofilm formation and aspartate protease production abilities than standard strain.The expression of virulence factors varies among strains isolated from different specimens,while there is no significant difference in the expression of virulence factors among strains isolated at different periods.Patients may have been infected with different MT types of C.parapsilosis in multiple sites during the same period.

OBJECTIVETo understand the environmental risk factors on attempted suicide, and to study the interaction between factors as gene polymorphism of catechol-O-methyltransferase (COMT) associated to attempted suicide.

METHODSPaired case-control study of 205 suicide attempters (89 male, 116 female) and molecular biological techniques were used to study the relation between gene polymorphism of COMT, environmental factors and the rate of attempted suicide. Controls were paired with cases according to the same gender, similar age (no more than 3 years) and in the same district.

RESULTSThere were no significant differences in gene types and gene frequency between case and control groups. Multivariate conditional logistic regression model analysis showed that COMT Val/Val 158/108, low education level, cigarette smoking, emotional conflicts, psychologic disorders and depression were risk factors of attempted suicide with OR values as 2.43 (95% CI: 1.10 - 5.40), 5.70 (95% CI: 1.88 - 17.27), 3.54 (95% CI: 1.02 - 12.36), 10.96 (95% CI: 4.74 - 25.34), 6.35 (95% CI: 1.68 - 24.05) and 11.30 (95% CI: 4.58 - 27.89) respectively. There was no first level interaction between any two risk factors.

CONCLUSIONThe study supported that low education level, cigarette smoking, affective conflicts, psychiatric disorders, depression were risk factors of attempted suicide and COMT Val/Val 158/108 was suspected to be a susceptible gene type of attempted suicide but needs further study. The study also suggested that 116 bp in gene atlas be possibly correlated to high activity of COMT.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Catechol O-Methyltransferase ; genetics ; China ; epidemiology ; Depression ; complications ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Smoking ; adverse effects ; Suicide, Attempted ; statistics & numerical data

Objective To investigate the impact of sarcopenia on muscle strength and exercise capacity in elderly patients with chronic heart failure (CHF).Methods One hundred and sixteen inpatient aged ≥65 years with CHF were enrolled in the study.General clinical characteristics were collected with questionnaire;the appendicular skeletal muscle mass (ASM) was assessed with body composition analyzer;the arm muscle strength was measured and a 6m-walk test was performed.The sarcopenia was evaluated according to the Asian Working Group for Sarcopenia (AWGS) criteria.Simple regression analysis and logistic regression were used to analyze the correlation between sarcopenia and decreased muscle strength,exercise capacity in elderly CHF patients.Results Among 116 elderly CHF patients,sarcopenia was confirmed in 42 patients(36.2％).Patients with sarcopenia had poorer cardiac function and lower muscle strength,ASM and 6m-walking speed than patients without sarcopenia did(allP＜0.05).Simple regression analysis showed positive correlation between ASM and muscle strength in both groups.Logistic regression analysis showed that after adjustment for confounders,ASM reduction was an independent risk factor for reducedexercisecapacity(OR=0.158,95％CI:0.055-0.455,P＜0.05).Conclusion Sarcopenia is a common complication in elderly patients with CHF and may lead to decreased muscle strength and exercise capacity.

Objective To investigate the association between sarcopenia and nutritional status among inpatients aged 80 years and over.Methods A total of 120 patients aged ≥80 years admitted into geriatrics department of a third-grade first-class hospital in Shandong province who met inclusion and exclusion criteria were enrolled from March 2017 to March 2018 in this cross-sectional study.According to diagnostic criteria of Asian Working Group for Sarcopenia,patients were divided into the sarcopenia group and the non-sarcopenia group.General clinical data were collected.Nutritional assessment was validated by using Mini Nutritional Assessment(MNA)and the determined nutritional indexes of serum levels of hemoglobin and albumin.Univariate and multivariate analysis were used to analyze the association between sarcopenia and nutritional status.Results Of 120 inpatients aged ≥ 80,28 cases(23.3％)had sarcopenia,including 20 men(71.4％)and 8 women(28.6％).The incidences of sarcomenia were 1.4％ (1 case)in patients with normal nutritional status and 100％ (6 cases)in patients with malnutrition.Compared with the non-sarcopenia group,the sarcopenia group had lower levels of body mass index,serum hemoglobin,serum albumin and MNA scores[(20.0 ± 2.2) kg/m2 vs.(25.4±3.0)kg/m2,(34.5±3.4)g/L vs.(38.6±3.5)g/L,(114.4± 14.0)g/L vs.(127.3± 14.8) g/L,(8.8 ± 1.9) score vs.(12.7 ± 1.4) score,P ＜ 0.05].After adjustment for all covariates,multivariate Logistic regression analysis showed that MNA score was an independent risk factor for sarcopenia(OR =0.118,95％CI:0.026-0.530).Conclusions The incidence of sarcopenia is high in hospitalized patients aged 80 years and older.MNA score is an independent risk factor for sarcopenia.

BackgroundThe proliferation of lens epithelial cellsLECs) following extracapsular cataract extraction is the biological basis of posterior capsular collagen and cataract formation.Disintegrin is certified to competitively bind the integrin with extracellular matrix and therefore prevent the posterior capsular opacification (PCO).But,its molecular mechanism is below understand.ObjectiveThe present study was to investigative the effects of disintegrin (kistrin) on the expression of collagen in lens posterior capsular.MethodsThe right eyes of 24 New Zealand white rabbits received transparent lens extracapsular enudeation and were randomly divided into two groups using random number table,0.2 ml of kistrin ( 80 mg/L) was intracapsularly injected at end of the operation in 12 eyes ( kistrin group) and the same volume of normal saline solution was used at the same way in other 12 operative eyes ( normal saline group).The PCO was graded in postoperative 1,3,5,7,14 days on Odrich' s criteria under the slit lamp.The lens section was prepared at 14 days and 3 months after operation.Haematoxylin and eosin stain was used to examine the proliferation of LECs in posterior capsule; Masson stain was used to observe the collagenous fiber formation in capsule bag,and the expression of collagen type Ⅳ was detected by immunochemistry.Results No significant difference in the number PCO eye was found in postoperative 14 days between normal saline group and kistrin group ( P =0.093 ).However,the eye numbers of 2-3 grades of PCO were significant increased in normal saline group compared with kistrin group in 1,2,3 months after operation( P=0.041,0.014,0.022).In the operative 14 days,staining and adhesion of LECs in posterior capsule were more in normal saline group than kistrin group,and the fibrocytes in capsule were evidently increased in normal saline group in 3 months.Masson stain certified that the blue stain was seen to be stronger and more in posterior capsule in normal saline group in comparison with kistrin group in 3 months after operation,and the immunochemistry showed that the gray values of collagen type Ⅳ in posterior capsule were significant lower in normal saline group compared with kistrin group in both 14 days and 3 months after operation (P=0.000,0.001 ).ConclusionsKistrin can suppresses the proliferation of LECs and collagen type Ⅳ on rabbit lens posterior capsular after transparent lens extracapsular enudeation.