Objective To construct eukaryotic expression vector encoding rat IMD gene and deliver it into rat renal tissue via ultrasound-mircobubbles. Methods IMD gene was inserted into pCDNA3.1 ( + )between Hind Ⅲ and EcoRI enzyme sites. The recombinant plasmid designated as IMD-pCDNA 3.1 wasconfirmed by restrictive enzyme digestion and sequencing. Eighteen male Wistar rats were randomized into 3groups, which were treated with no transfection, empty vector transfection and IMD transfection, respectively, in renal tissue via ultrasound-microbubbles. RT-PCR and Western blotting were applied to detect the expression level of IMD. Results Enzyme- digestion and sequencing data showed that IMD-pCDNA 3.1 was correctly constructed. The differences in ALT, AST, BUN and SCr were not significant; No obvious damage in the glomerular, tubular and interstitial was observed in all the treated groups;Compared with non-transfection group and empty vector-transfection group, IMD mRNA and protein expression in IMD transgenic renal tissue were significantly increased. Conclusion IMD-pCDNA 3.1 expression vector was successfully constructed and well expressed in rat kidney.

Objective To investigate the diagnostic and prognostic value of serum soluble CD163 (sCD163 )and the positive rate of membrane -bound CD163 (mCD163 )in peripheral blood mononuclear cells (PBMC)in children with infection -associated hemophagocytic syndrome (IAHS).Methods Between July 2012 and June 2016,26 pediatric patients with IAHS (IAHS group)and 28 pediatric patients with sepsis(sepsis group)admitted to Children′s Hospital Affiliated to Shanghai Jiaotong University were selected,and 20 healthy children were taken as healthy control group. Sandwich enzyme linked immunosorbent assay was used to detect serum sCD163 .The population of circulating mCD163 positive monocytes was determined by using flow cytometry.Receiver operating characteristic (ROC)curves were used to evaluate the diagnostic and prognostic values of sCD163 and mCD163 in children with IAHS compared with the diagnos-tic and prognostic values of plasma ferritin,and so on.Results The serum levels of sCD163 in patients of IAHS group, sepsis group and healthy control group were (1264 ±538)mg/L,(862 ±332)mg/L,(610 ±316)mg/L,respective-ly.And the population of mCD163 -positive PBMC in patients of IAHS group,sepsis group and healthy control group was (88.3 ±9.7)%,(68.5 ±18.3)%,(28.9 ±5.2)%,respectively.Both serum sCD163 and the population of mCD163 -positive PBMC were significantly higher in IAHS group compared with those of sepsis group (t =2.031 ,P =0.048;t =3.191 ,P =0.002,respectively).The serum sCD163 and population of mCD163 -positive PBMC in sepsis group were higher than controls (t =3.848,P =0.002;t =4.049,P =0.000,respectively).Moreover,the areas under the ROC curve (AUC)for the mCD163 ,sCD163 ,were 0.853(P =0.013),0.762(P =0.004),0.755(P =0.049),respec-tively.mCD163 at a cutoff of 83.7% had a high diagnosis sensitivity (81 .8%)and specificity (72.4%).The optimal cutoff values of sCD163 and ferritin for predicting IAHS was 888 mg/L (sensitivity 66.7% and specificity 63.3%)and 2880 μg/L (sensitivity 80.0% and specificity 54.5%).In addition,the serum level of sCD163 and the population of mCD163 -positive PBMCs were significantly increased in acute phase and decreased in recovery phase[(1553 ±542) mg/L vs.(866 ±92)mg/L,(91 .0 ±6.4)% vs.(79.0 ±4.6)%,t =2.450,χ2 =3.419,P =0.036,0.007]in IAHS group.Furthermore,subgroup analysis indicated that the serum level of sCD163 and the population of mCD163 -positive PBMCs were significantly higher in dead patients than those in survived patients [(1748.91 ±518.17)mg/L vs. (909.69 ±171 .35)mg/L,t =3.070,P =0.011 ;(93.50 ±8.42)% vs.(77.30 ±3.28)%,χ2 =3.005,P =0.024, respectively].Conclusion Serum sCD163 and the population of mCD163 -positive PMSCs are specific and validity bio-markers for early diagnosis of IAHS,which also are associated with treatment response assessment and prognostic analy-sis in IAHS.

Objective To investigate the protective effect of intermedin(IMD)on renal ischemia reperfusion injury(IRI)and its mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups: control group, IRI group, empty plasmid group and IMD group. After remove of right kidney, plasmid was transfected into the kidney by ultrasonic microbubbles technology, and IRI model was made after 1 week. Renal pathology was observed by PAS staining. Renal tissue superoxide dismutase(SOD), myeloperoxidase(MPO), caspase-3 activity, and malondialdehyde(MDA)content were detected by colorimetric method. The intercellular cell adhesion molecule-1(ICAM-1), endothelin 1(ET-1)and P-selection expression of renal tissue were detected by immunohistochemical method. Apoptosis of renal tubular cell was detected by TUNEL.Results Compared with control group, tubulointerstitial pathological injury was significant aggravated in IRI group(P<0.01);compared with IRI group, IMD pretreatment significantly alleviated the degree of renal injury(P<0.01). Compared with control group, in IRI group, SOD activity was significantly decreased(P<0.05), MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 were increased significantly(all P< 0.01). Compared with IRI group, IMD pretreatment significantly increased SOD activity(P <0.05), decreased the MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 (all P<0.01). The apoptosis rate of renal tubular epithelial cells in IRI group was significantly higher than that in control group(34.83%±8.75% vs 3.33%±0.47%, P<0.01), while the apoptosis rate of IMD group(20.67%±7.71%)was significantly lower than that of IRI group. There was no difference of above indexes between empty plasmid group and IRI group. Conclusions IMD pretreatment protects against renal IRI. The mechanism may be at least partly related to the clearance of oxygen free radicals, the improvement of lipid peroxidation, inflammatory cell infiltration and cell apoptosis, leading to the decrease of the production of reactive oxygen species caused by oxidative stress.

Objective To investigate the effect of intermedin (IMD) on renal ischemia/ reperfusion (I/R) injury after the up-regulation of IMD. Methods A total of 24 healthy Wistar male rats were randomly divided into four groups,sham-operated group,I/R group,IMD gene transfection +I/R group and empty plamid +I/R group.All the animals were killed at the end of 24 h of reperfusion.Histological changes and renal function were estimated.The expression and site of IMD were determined by Immunohistochemistry method,semi-quantitative RT-PCR and Western blotting.The protein expressions of endothelin 1 (ET-1),tumor necrosis factor αt (TNF-α) were detected by Western blotting. Results Compared with sham-operated group,tubulointerstitial pathological injury was significant aggravated in I/R group (7.6±2.3) and empty plamid +I/R group (7.0±1.8),and such injury was improved in IMD+I/R group (1.5±0.8) (P＜0.05).Compared with I/R group and empty plamid +I/R group,the renal dysfunction of IMD +I/R group was obviously lessened [BUN:(7.73±1.03) mmol/L vs (10.13±2.14) mmol/L,(9.77±1.92) mmol/L; Scr:(58.50±3.27) μmol/L vs (80.33±7.15) μmol/L, (75.67±7.58) μmol/L,all P＜0.05].IMD expression was weak in the plasma of tubulointerstitial cells in sham-operated group,and was up-regulated in I/R group. Compared with I/R group, immunohistochemical IMD expression increased obviously (262.03±67.89 vs 175.57±48.06,P＜0.01).The mRNA expression of IMD in IMD+I/R group was up-regulated significantly by 60.7％,66.1％ and the protein expression of IMD in IMD+I/R group increased significantly by 51.4％,55.9％ as compared to I/R and empty plasmid +I/R group.Meanwhile,the protein expressions of ET-1 and TNF-αt in IMD+I/R group were obviously lower compared with those in I/R group (ET-1:0.08±0.02 vs 0.17±0.02; TNF-α:0.21±0.04 vs 0.35± 0.02,all P＜0.05). Conclusion IMD gene transfected into kidneys of rats prior to I/R surgery can attenuate the over-expressions of both ET-1 and TNF-o in I/R injured rat kidneys as well as the damages to the structure and function of the kidneys.

OBJECTIVETo study the effects of rhizoma sparganii and radices zedoariae on hepatic fibrosis.

METHODThe rat immunohepatic fibrosis model was made by intraperitoneal injection of porcine serum and treated with rhizoma sparganii and radices zedoariae. The ALT, GGT, TP, ALb, A/G, IVC, LN, HA and the pathological change of the liver were observed.

RESULTRhizoma sparganii and radices zedoariae could increase TP, ALb, A/G, decrease ALT, GGT, IVC, LN, HA and improve the pathological change.

CONCLUSIONRhizoma sparganii and radices zedoariae can protect hepatic cells, alleviate degeneration and necrosis, recover structure and function, and reduce the proliferation of fibrous tissue.

Animals ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Liver Cirrhosis ; drug therapy ; pathology ; Magnoliopsida ; chemistry ; Male ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Rhizome ; chemistry

OBJECTIVETo investigate the relationship between the changes of the L-type calcium current (I(Ca, L)) and the calcium-activated transient outward chloride current (I(Cl, Ca)), and the repolarization characteristic of action potential in phase 1 under isoprenaline (ISO) stimulation in atrium myocytes of rabbit.

METHODSAtrium myocytes were obtained by enzymatic dissociation from a section of atrial free wall. The membrane currents and action potential were recorded by the whole-cell patch-clamp technique.

RESULTSAfter recording I(Ca, L), atrium myocytes were perfused with ISO (1 micromol/L) immediately. Five minutes later, a transient outward current (I(to)) was significantly induced, and the peak of I(to) was gradually increased while I(Ca, L) gradually decreased with increasing in clamp voltage. The I(to) was resistant to 4-AP (3 mmol/L) but sensitive to DIDS (150 micromol/L, Cl(-) channel blocker). This current was blocked by CdCl(2) (200 micromol/L, Ca(2+) channel blocker). The elicited rate of I(to) was 91.67% (P < 0.05). (2) The shape of AP was like an inverse triangle with no plateau in Phase 2 after ISO (1 micromol/L) perfusion. Moreover, compared to the parameters of control group, APD(50) and APD(90) were significantly shortened from (65.4 +/- 4.2) ms and (95.8 +/- 3.8) ms to (12.8 +/- 3.8) ms and (27.0 +/- 4.7) ms, and reduced to 80.46% and 71.87%, respectively (P < 0.01, n = 12). 4-AP (3 mmol/L) had on obvious effect on the shape of AP, however, the plateau of AP in phase 2 was recovered by DIDS (150 micromol/L) perfusion, APD(50) and APD(90) were (41.1 +/- 4.5) ms and (79.6 +/- 3.4) ms respectively. Compared to the parameters of control group, there were no significant differences (P > 0.05, n = 12). These results indicated that ionic transport were changed by ISO perfusion in atrium myocytes and I(to) played an important role in the phase 1 repolarization of AP.

CONCLUSIONSBefore ISO administration, we could only observe I(Ca, L) in atrium myocytes of rabbit. After isoproterenol intervention, certain intracellular ionic consistency and membrane ionic channels were changed. Calcium activated chloride channel and I(to2) revealed obvious predominance which shorten APD significantly. Action potential showed a triangle with no plateau, suggesting an electrical remodeling in atrium myocytes. The remodeling of ionic channel is related possibly with the opening of Ca(2+)-activated Cl(-) current, which maybe the electrophysiological base of reentrant atrial tachycardia.

Animals ; Calcium ; metabolism ; Calcium Channels, L-Type ; metabolism ; Calcium Signaling ; Cells, Cultured ; Chloride Channels ; metabolism ; Heart Atria ; cytology ; metabolism ; Ion Transport ; Isoproterenol ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; Patch-Clamp Techniques ; Rabbits

OBJECTIVETo elucidate the relationship between surgical time and postoperative complications in senile patients with hip fractures, and try to find out other factors which are related to these complications.

METHODSSixty-two patients, 28 males aged from 65 to 72 years with a mean age of 76.3 years and 34 females aged from 65 to 95 years with a mean age of 78.1 years, who had undergone orthopedic surgery because of hip fractures, were enrolled in a retrospective cohort study. The surgical time and pattern, the type of fracture, preoperative comorbidities, American Society of Anesthesiologists (ASA) score and the volume of blood transfusion during operation were obtained from these patients who were followed up by telephone calls for postoperative complications. All the patients were followed up at least for 1 year and were divided into subgroups according to their clinical characteristics and the results were analyzed by the Statistical Analysis System software.

RESULTSThere was no significant difference in the morbidity of postoperative complications with the gender, age, surgical time and pattern, or ASA score. There was significant difference in the morbidity of postoperative complications related to preoperative comorbidities and the volume of blood transfusion. There was a significant causality between preoperative comorbidities and postoperative complications. The morbidity of postoperative complications was 1.651 times higher in patients with preoperative comorbidities than those without.

CONCLUSIONSThere is no relationship between the surgical time and postoperative complications in senile patients who received surgery for hip fracture within 1 year. No correlation is found between the postoperative complications and gender, age, type of fracture, surgical pattern, ASA score and the volume of blood transfusion. Preoperative comorbidities are an independent predictor for postoperative complications.

Aged ; Aged, 80 and over ; Cohort Studies ; Female ; Hip Fractures ; surgery ; Humans ; Logistic Models ; Male ; Morbidity ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Time Factors

Adult ; Bilirubin ; blood ; Cost-Benefit Analysis ; DNA, Viral ; blood ; Female ; Humans ; Liver Failure ; economics ; mortality ; therapy ; Male ; Middle Aged ; Plasma Exchange ; economics ; methods ; Prothrombin ; metabolism ; Retrospective Studies ; Severity of Illness Index ; Survival Rate ; Treatment Outcome

Objective To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneously determining five compounds in Curcumae Aromaticae Radix.Methods An HPLC method was developed as QAMS to determine curcuma diol,ocathydro-1,4-dihydroxy-1,4-dimethyl-7-(propan-2-ylidene)azulen-5(1H)-one,original curcumol and curcumin in Curcumae Aromaticae Radix,using curdione as intermal reference substance,and the relative correction factor (RCF) of the four components was determined by HPLC with good reproducibility.Their contents in 10 batches of samples,collected from different areas,were determined by both external standard method and QAMS.Result No significant differences were found in the quantitative results of four compounds in 10 batches of Curcumae Aromaticae Radix determined by external standard method and QAMS.Conclusion It is feasible and suitable to evaluate the quality of Curcumae Aromaticae Radix by QAMS.

Objective To observe the protective effects of a mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride (CCl_4). Methods The model of rat acute liver damage was established by injection of CCl_4 2 ml/kg into the abdominal cavity.The rat models were treated respectively by the mixture for protecting liver and supplementing stomach 30 g?kg~(-1)?d~(-1),the polyene phosphatide acid radical choline capsule [Yi Shanfu (易善复), 180 mg?kg~(-1)?d~(-1)],the glycyrrhizic acid diaminogen capsule [Gan Lixin (甘利欣),30 mg?kg~(-1)?d~(-1)] infused into the stomach.The activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected.In the mean time,the liver pathological changes were observed,the degree of liver cell necrosis was evaluated,and the rat mortality was noted in various groups of treatment.Results The values of ALT,AST and the score of liver cell necrosis in the group treated with the mixture for protecting liver and supplementing stomach [(1.168?1.066) kU/L,(1.845?2.212) kU/L,(0.56?0.53) score] were significantly lower than those in the model group [(4.982?3.502) kU/L,(7.030?3.616) kU/L, (1.38?0.92) scores],and all the differences being statistically significant (all P