Objective To retrospectively analyze the serum enzymes in childhood amputees as a result of earthquake,and to discuss their clinical significance.Methods From 150 children amputees who were victims of Sichuan Wenchuan earthquake in 2008 and Sichuan Lushan earthquake in 2013,45 cases with complete records of serum enzymes examinations were reviewed retrospectively.They were divided into three groups:amputation group (n=6),fasciotomy decompression group (n =5),general trauma without injury to extremity group (n =34).Serum enzyme examination data were compared for statistical analysis to find the difference among groups.Ten children who were not victims of earthquake were selected from department of orthopaedics to serve as controls,and 20 adult amputees as a result of earthquake served as another control group.Results There were significant differences in injury severity scale (ISS) and the contents of all serum enzymes,including aspartate transaminase (AST),alanine aminotransferase (ALT),creatine kinase (CK),lactate dehydrogenase (LDH),and hydroxybutyrate dehydrogenase (HBDH),between the amputation group and other patients (FISS=15.474,P＜0.001 ; FAST=10.770,P＜0.001 ; FALT=12.799,P＜0.001 ; FcK=12.848,P＜0.001; FUDH=13.126,P＜0.001; FHBDH=13.186,P＜0.001),and the difference in amputation group was even more significant.AST,LDH,HBDH in childhood amputees group were significantly higher than those in adult amputation group.The contents of ALT and CK were slightly increased.Serum enzyme contents were found to be significantly helpful for prediction of disease condition and prognosis.It was also found that CK was extremely helpful in assessing the degree of illness in patients with severe trauma,especially in patients complicated by severe soft tissue injury.In all the groups,of patients,it was found that CK rose from (129 ± 62) U/L in non-earthquake induced trauma group to (44 208 ± 39 788) U/L in earthquake amputation group,and it was thus increased more than 300 times.Its highest value even reached 117 513 U/L,which was more than 840 times of the normal.If a timely amputation or muscle compartment decompression was performed,CK might decline rapidly down to the normal value.Conclusions The comprehensive and continuous assessment of serum enzymes is mandatory during the treatment of children with acute trauma.It is of important clinical significance to correctly judge the condition and to determine optional treatment measures.

ObjectiveTo investigate the expression and activity of Rho-associated kinase 1 (ROCK1) in renal tissues of uninephrectomized diabetic rats,and to explore the possible mechanism of renal protection of spironolactone.Methods The model rats were established by a single intraperitoneal injection of streptozotocin(STZ) after uninephrectomy and randomly divided into sham operation group(N),uninephrectomized control group?,uninephrectomized diabetic group (D),and spironolactone treated group (S).Four and 8 weeks later,biochemical indexes and renal morphology were detected.Expressions of ROCK1 and connect tissue growth factor (CTGF) were examined by immunohistochemistry and real-time PCR.Protein expression of p-MYPT1 was examined by Westem blotting.Results After 4 and 8 weeks,compared with group N and C,blood glucose,systolic blood pressure,24-h urinary protein,kidney weight/body weight (KWI) were significantly increased (P＜0.01),and extracellular matrix proliferation and basement membrane thickening were found in group D.After 8 weeks in group D,Alb significantly decreased and Scr significantly increased (P＜0.05).In group D,protein and mRNA expression of ROCK1 and CTGF increased significantly and protein expression of p-MYPT1 increased significantly as well with time.Treatment with spironolactone could partially reverse those changes. ConclusionsExpression and activity of ROCK1increase in renal tissues of uninephrectomized diabetic rats and are positively correlated with the expression of CTGF.Spironolactone can protect the kidney of diabetic rats in early stage probably through decreasing the expression of ROCK1,CTGF and inhibiting the activation of ROCK1.

10.3969/j.issn.1000-3606.2013.06.004

A RP-HPLC method was established for simultaneous determination of phellodendrine hydrochloride (PH1), magnoflorine hydrochloride (MH), jatrorrhizine hydrochloride (JH), palmatine hydrochloride (PH2) and berberine hydrochloride (BH) in Phellodendri Chinensis Cortex by using ionic liquids as mobile phase additives. The separation was performed on a Kromasil C18 (4.6 mm x 250 mm, 5 μm) coupled with ultraviolet (UV) detection. The effect of extraction solvent, detection wavelength, length of alkyl chain on different imidazolium ionic liquids and concentration of ionic liquids on the separation and determination of alkaloids were investigated. Ionic liquid, [BMIm] BF4, can obviously improve the resolution and peak shape. This ILs-HPLC method is simple, rapid, and reliable, which can be used for determination of alkaloids in Phellodenddri Chinensis Cortex.
Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Phellodendron ; chemistry

A full-length cDNA sequence encoding for the precursor of a venom peptide (named BmKCT) with homology to chlorotoxin has been isolated from a cDNA library made from the venom glands of the Chinese Scorpion Buthus martensii Karsch. The sequence of BmKCT is similar (68 % identities) to that of chlorotoxin isolated from Leiurus quinquestriatus quinquestriatus. To understand the biological function of BmKCT, this peptide was expressed using pGEX expression system and purified using GST affinity column and gel filtration.Whole cell patch-clamping recording showed that BmKCT could significantly inhibit chloride currents of gliomas cells, and the inhibitory effect was reversible. These results suggested that BmKCT might belong to the class of short chain toxins blocking the chloride ion channels.

Objective To investigate the effect of Caveolin-1 on extracellular regulated protein kinases of rabbit carotid artery anastomotic restenosis.Methods 40 New Zealand white rabbits were randomly divided into normal control group,carotid artery end to end anastomosis surgical group,empty vector transfection on the site of anastomosis group and Caveolin-1 transfected group.Left carotid artery endto-end anastomosis was performed,and the mixture of Caveolin-1 plasmid and liposome lipofectin 2000 (transfected group) or empty plasmid and lipofectin 2000 mixture (empty vector group) were transfected on anastomosis.Specimens were taken at 7 d after surgery for Western blot and RT-PCR to detect the expression of protein and mRNA.Specimens were taken for HE staining at 28 d to observe the proliferation of intima,and measured the ratio of intima/media area by Image-Pro Plus 6.0 software.Results Compared with surgical group,the ratio of intima/media area in Caveolin-1 transfected group decreased by about 50％.Compared with surgical group and empty vector group,the Caveolin-1 mRNA expression and protein activity significantly increased (t =36.59,P ＜ 0.01) ; the ERK1/2 mRNA expression and protein activity significantly decreased on rabbit carotid artery anastomotic site in Caveolin-1 transfected group (t =32.64and 7.27,P ＜ 0.01).Conclusions Caveolin-1 inhibits anastomotic restenosis possibly by regulating the activation of ERK.

Objective To investigate the effect of caveolin-1 on rabbit carotid artery anastomotic stenosis and its relationship with TNF-α.Methods 40 New Zealand white rabbits were randomly divided into normal group,surgical group,empty vector group and caveolin-1 transfecting group.Carotid artery end-to-end anastomosis was done in the rabbits except these in normal group.Five specimens were randomly taken on day 7 after surgery for Westem blot to detect the expression of caveolin-1 and TNF-α; The rest specimens were taken for HE staining at fourth week.The ratio of intima/media area were measured by Image-Pro Plus 6.0 software in order to observe the proliferation of intima.Results Compared with normal group,in surgical group intimal proliferation was significant,the intima/media ratio was significantly higher (P ＜ 0.05) ; Compared with surgical group,in caveolin-1 transfected group neointimal proliferation was not obvious,the intima/media ratio decreased (P ＜ 0.05).Western blot showed that:compared with the surgical group,caveolin-1 expression in transfected group was significantly higher (P ＜ 0.05) ; compared with normal group,the TNF-α expression in surgical group increased (t:41.28,P ＜ 0.05) ; Compared with surgical group,TNF-α expression in transfected group decreased significantly (t:36.37,P ＜ 0.05).Conclusions Caveolin-1 inhibits vascular anastomotic stenosis,possibly by down-regulating TNF-α expression.

Acetaminophen ; metabolism ; Animals ; Cells, Cultured ; Drug Interactions ; Flavonoids ; metabolism ; Hepatocytes ; metabolism ; Humans

Objective To develope a new Real-time quantitative PCR assay using SYBR green as fluorescence reporter, which is rapid, specific, sensitive, cheap and accurate for the detection of mycoplasma pneumoniae(MP), and evaluated its clinical application value.Methods The sequence of the 23S rRNA gene in MP type strain FH was selected as amplified regions, and specific primers were designed.Then the related plasmids were extracted as standards,and the absolute quantitative standard curve was established.The sensitivity ,specificity of the fluorescence quantitative PCR assay was compared with the nest-PCR and kit;To calculate correlation coefficient, coincidence rate and kappa coefficient, clinical samples were detected using above-mentioned methods and cultivation,respectively.Results The detection sensitivity of the new real-time PCR and nest-PCR was 10 copies of FH DNA,while the kit 100 copies.In the specificity tests,the MP sample was positive,while mycoplasma hominis and other four bacteria were all negative.We applied this real-time PCR assay ,nest-PCR, kit and cultivation to 182 clinical specimens, and the detection rates were 55.49％, 52.75％, 47.25％ and 39.01％ ,respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and nest-PCR were 89.6％ ,0.790, respectively;while those of the new method and cultivation were 83.5 ％ ,0.678, respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and the kit were 89.6％ ,0.792,respectively;and the correlation coefficient of these two methods was 0.923,P ＜ 0.001.Conclusion Compared with other methods, the new real-time PCR assay could be used to detect mycoplasma pneumoniae quickly and economically, with high sensitivity and specificity ,revealing great utility value on varied instrumentation platforms.

Objective To evaluate the rehabilitation effects of walk exercise training on the heart function rehabilitation in the patients with heart failure with preserved left ventricular ejection fraction (HFPEF) .Methods A total of 142 cases of HFPEF were randomly assigned to 2 groups on the basis of medication therapy ;the control group (70 cases) was given only routine medica‐tion therapy without the exercise prescription;On the bases of control group ,the exercise group (72 cases) conducted the rehabilita‐tion training according to the exercise prescription .After 24 weeks follow up ,6 min walking distance ,plasma NT‐proBNP level and left ventricular diastolic function were compared between the two groups .Results The 6 min walking distance after walk exercise training in the exercise group was significantly increased compared with before walk exercise training and the control group ,while the level of NT‐proBNP was significant decreased(P<0 .01) .Conclusion The walking exercise training mode according to the ex‐ercise prescription can significantly improve the exercise tolerance in the HFPEF .