Purpose:To study the effect of early restrictive fluid resuscitation (EFR) on inflammatory and immune factors in patients with severe pelvic fracture (SPF).Methods:A total of 174 SPF patients in the Department of Orthopaedics,the First Affiliated Hospital of Chengdu Medical College from July 2015 to June 2018 were involved in this study and divided into EFR group (n =87) and control group (n =87) using the random number table method.Conventional fluid resuscitation (CFR) was performed in control group,and EFR was performed in EFR group.The incidences of acute respiratory distress syndrome (ARDS) and multiple organ dysfunction syndrome (MODS) during rescue,successful rescue rate,blood transfusion volume,fluid input,and resuscitation time were compared between the two groups.The parameters including prothrombin time (PT),hematocrit (HCT),platelet (PLT) and blood lactate (BL) at the 4th hour after fluid resuscitation were recorded.The levels of inflammatory factors (TNF-α,IL-6,CRP) and immune factors (CD3+,CD4+,CD8+,CD4+/CD8+) were compared between the two groups before treatment and 7 days after treatment.The revised acute physiologic and chronic health evaluation system and the sequential organ failure assessment scores were adopted for evaluation before treatment and 7 days after treatment.Results:The incidences of ARDS and MODS during rescue in EFR group were significantly lower than those in control group (p=0.015 and 0.010 respectively),and the successful rescue rate in EFR group was significantly higher than that in control group (p =0.011).The blood transfusion volume,fluid input,resuscitation time in EFR group were significantly lower than those in control group (p =0.016,0.002 and 0.001 respectively).At the 4th hour after fluid resuscitation,PTand BL in EFR group were significantly lower than those in control group (p =0.021 and 0.003 respectively),while HCT and PLT in EFR group were significantly higher than those in control group (p =0.016 and 0.021 respectively).On day 7 after treatment,TNF-α,IL-6,CRP and CD8+ in EFR group were significantly lower than those in control group (p =0.003,0.004,0.007 and 0.003 respectively),while CD3+,CD4+ and CD4+/CD8+ in EFR group were significantly higher than those in control group (p =0.004,0.000,0.007 respectively).On day 7 after treatment,the revised acute physiologic and chronic health evaluation (APACHE) system and the sequential organ failure assessment (SOFA) scores in EFR group were significantly lower than those in control group.Conclusion:EFR can effectively eliminate inflammatory factors,improve immune function,maintain the stability of blood components,reduce the incidences of ARDS and MODS,and elevate the successful rescue rate in patients with SPF.

Culture environment is the key factor in the construction of dermal skin.It was investigated that the effects of the culture methods,including the static culture and spinner flask culture,and stir speeds on the cells proliferation,metabolism and distribution within collagenchitosan sponges.A higher cell density and specific growth rate was obtained with spinner flask culture versus static culture,especially,the 80 r/min spinner flask culture.The cell distribution in dermal substitutes from stirred culture system was more uniform than static culture,as well as that with increase of stir speeds in spinner flask.In summary,the spinner flasks culture with proper stir speed shows promise for the construction of dermal substitutes in vitro.

Objective To investigate the simulation exercise on emergency timeliness of pre-hospital trauma patients with the radius of ≤6 km. Methods A retrospective control study was taken, and 170 cases of trauma patients before and after exercise, respectively, were randomly selected. Pre-hospital first aid implementation of the pre-hospital time, first-aid radius, the actual distance first aid, and emergency response time were compared. Results Time of first aid, first aid radius, the actual distance aid were no statistically significant (P ＞ 0. 05); the pre-hospital first aid, emergency response time were all statistically significant (P ＜ 0.05). Conclusions Pre-hospital trauma simulation exercises had a strong guide to the pre-hospital trauma patients with the radius of ≤ 6KM emergency, could improve the treatment time, and increase the implementation of effective rescue measures and shorten emergency response time.

OBJECTIVETo discuss the reverse effect of Yinchenhao decoction(YCHD) in dimethyl nitrosamine (DMN)-induced hepatic fibrosis in rats.

METHODThe rat hepatic fibrosis model was established through the intraperitoneal injection with 1% dimethyl nitrosamine (DMN) with a dose of 1.0 mL x kg(-1) x d(-1) for consecutively three weeks, once for the first three days of each. The rats were randomly divided into six groups: the silymarin positive control group (50.0 mg x kg(-1) x d(-1), YCHD high (20.0 g x kg(-1) d(-1)), middle (8.0 g x kg(-1) x d(-1)) and low (3.2 g x kg(-1) x d(-1)) dose groups, the model group and the normal control group. The model group and the normal control group were orally administered with normal saline for consecutively five weeks. The pathologic changes in liver tissues were observed by HE staining. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), g-glutamyltransferase (g-GGT), hyaluronic acid (HA), laminin (LN), collagen type IV (CIV) and type III procollagen amino terminal peptide (PIIINP) in serum were determined. The metabolite profiling of amino acid and the content of hydroxyproline in liver tissues were also measured.

RESULTCompared with the model group, YCHD high and middle dose groups could significantly reverse the pathologic changes in liver tissues of rats. YCHD could reduce the levels of ALT, AST, gamma-GGT, HA, LN, CIV, PIIINP in serum and the content of hydroxyproline in liver tissues in a dose-dependent manner, and altered the metabolite profiling of amino acid in rat liver tissues.

CONCLUSIONYCHD has the effect in reversing dimethyl nitrosamine induced hepatic fibrosis in rats.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Collagen Type IV ; metabolism ; Dimethylnitrosamine ; adverse effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hydroxyproline ; metabolism ; Liver ; drug effects ; enzymology ; metabolism ; Liver Cirrhosis ; chemically induced ; drug therapy ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley

Objective To observe the effect of different thyroid hormone level on the expression of synaptotagmin Ⅰ(Syt Ⅰ) in adult rat hippocampus. Methods All 28 adult male SD rats were assigned randomly into hypothyroid, hyperthyroid and control group, hypothyroid group was established by daily intraperitoneal injections with propylthiou raci(PTU, 10.0 mg/kg body weight) for 6 weeks and hyperthyroid group with L-Thyroxine (L-T4, 0.5 mg/kg body weight) for 3 weeks. Radioimmunity method was used to assay the levels of serum T3 and T4, immunohistochemical S-P technology to assay the levels of Syt Ⅰ protein in hippoeampus CA1, CA3 and dentate gyrus (DG). The layers analyzed in the different subfields include the polymorphic cell layer(the stratum oriens, SO), pyramidal cell layer(PCL), stratum radiatum (SR), lacunosum-molecular layer (SLM) in CA1 and CA3, granular cell layer(GL) and molecular layer(ML) in DG. Results The levels of serum T3 and T4[(0.34±0.12), (41.03± 11.37)nmol/L]in the hypothyroid rats were significantly lower than those in the control group[(0.65±0.15), (55.20±10.68)nmol/L, P < 0.01 or < 0.05], and the positive granule of Syt Ⅰ was significantly lower in PCL and SR of CA1 and CA3, GL of DG. The average optical value responsible for Syt Ⅰ immunoreactivity was obviously reduced in SO(0.048±0.007), PCL(0.299±0.035), SR(0.042±0.007), SLM(0.038±0.006) of CA1, PCL(0.085± 0.019), SR(0.040±0.011), SLM (0.038±0.006) of CA3, GL (0.076±0.019) of DG than normal controls (0.068± 0.014, 0.376±0.053, 0.053±0.008,0.056±0.009,0.118±0.026,0.052±0.010,0.053±0.009,0.099±0.015; P< 0.01 or < 0.05). Serum T3 and T4 levels [(1.43±0.30), (157.18±19.95)nmol/L]of hyperthyroid rats were significantly higher than those of control group(P < 0.01). The value was reduced in PCL(0.322±0.050), SR(0.039±0.006), SLM (0.042±0.006) of CA1, PCL(0.098±0.034), SR(0.046±0.013), SLM(0.046±0.010) of CA3 and GL(0.085± 0.024), ML (0.042±0.009) of DG (P < 0.05 or < 0.01). Conclusion Adult-onset of hypothyroidism and hyperthyroidism can reversibly decrease the expression of Syt Ⅰ in CA1, CA3 and DG regions of hippocampus.

Hypodermin C (HC) cDNA was amplified from recombinant pGEM - T/HC, cloned in frame with the signal sequence in yeast vector pPIC9k. The plasmid was linerarized and transformed into Pichia pastoris GS115 strain by electroporation method. Recombinant strain was screened by G418 resistant, and further confirmed by PCR. The recombinant strain which contains insert was induced in the medium containing 0.5% methanol. The supernatant was collected and then purified by anion exchange chromatography. SDS-PAGE indicated that the target protein is around 28kD. Western-blot showed it can react with rabbit-anti HC serum. Gelatin substrate SDS-PAGE displayed it had enzyme activity. Provided a method to produce enough antigens for carrying out extensive immunological analyses.
Animals ; Blotting, Western ; Cell Line ; Chromatography, Ion Exchange ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Gelatin ; metabolism ; Gene Expression ; Immune Sera ; immunology ; Pichia ; genetics ; Plasmids ; genetics ; Rabbits ; Recombinant Proteins ; isolation & purification ; metabolism ; Serine Endopeptidases ; genetics ; immunology ; metabolism ; Substrate Specificity ; Transformation, Genetic

Female ; Humans ; Infant ; Propionic Acidemia ; diagnosis

Objective To develop a high-throughput clinical method on drag-resistance gene mutations of HBV using MALDI-TOF-MS. Method Using MassArray Assay Design software designed the iPLEX primers and followed the iPLEX instruction for amplification, SAP reaction, primer extenction, desalination, dispensing, MALDI-TOF-MS screening and data analysis of the gene mutation locus. 138 serum samples of chronic HBV patients with single drug-resistance or multiple drug-resistance on Lamivudin, adefovi, Entecavir were detected. Result The HBV gene mutation platform was successfully developed and applied on the high-throughput dectection of clinical serum samples. It was also a high throughput assay which could be used to detect for more than 138 samples once. The MALDI-TOF-MS technology and the DNA sequencing simultaneously examine 33 samples, in which result of 10 sample is inconsistent, the including 2 samples by MALDI-TOF-MS technology has not tested, 1 sample has 2 inconsistent mutations. Conclusion Detction of HBV gene mutations using MALDI-TOF-MS is highly-sensitive, highly-accuate, high-thoughput,fast achieved and suithle to use in the diagnosis and monitoring of HBV.

OBJECTIVETo express and purify Schistosoma japonicum ribosomal protein S4(SjRPS4) in Escherichia coli, and assess its value in immunodiagnosis of Schistosomiasis japonica.

METHODSGene fragment of SjRPS4 was amplified by screening the cercaria cDNA library of Schistosoma japonicum. The target gene was cloned into the expressive vector pQE30 and transformed into E. coli M15. The recombinant protein expression was induced by isopropylthio-β-D-galactoside (IPTG). This fusion protein was purified by Ni(2+)-NTA chromatography and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and ELISA.

RESULTSThe plasmid pQE30/SjRPS4 was constructed successfully and expressed a SjRPS4 fusion protein in E. coli as showing a single special band on SDS-PAGE gel at Mr 30 × 10(3) position. It reached a purity of above 90% after purification. The Western blot result confirmed that the recombinant protein could specifically react with the serum samples from patients of schistosomiasis. Detecting the serum of Schistosomiasis japonica patients by ELISA, the sensitivity and specificity of the ELISA method were 90.91% (70/77) and 92.59% (25/27), the positive rate of recombinant protein expression was 67.30% (70/104). There was no cross-reaction with paragonimiasis patients' serum.

CONCLUSIONProtein SjRPS4 was successfully cloned and expressed, and it was confirmed that SjRPS4 antibodies were valuable in the diagnosis of Schistosomiasis japonica.

Amino Acid Sequence ; Animals ; Antibodies, Helminth ; blood ; Antigens, Helminth ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Library ; Humans ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; genetics ; Ribosomal Proteins ; genetics ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; diagnosis ; genetics ; Sensitivity and Specificity

OBJECTIVETo study the effect of several factors on the quantity of hypericin in H. perforatum callus.

METHODHigh efficiency liquid phase chromatography and plant tissue culture were applied.

RESULT AND CONCLUSIONWhen the ratio of nitro-nitrogen to amina-nitrogen is 3:1, the callus biomass is 1.6-fold and the synthetic mass of hypericin rises. 0.1-0.20 mg x L(-1) mannose improves the content of total hypericin. The addition of PVP or PVPP can promote improvement of the growth and biosynthesis of callus.

Culture Media ; Hypericum ; growth & development ; metabolism ; Mannose ; pharmacology ; Nitrogen ; pharmacology ; Perylene ; analogs & derivatives ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Povidone ; analogs & derivatives ; pharmacology ; Tissue Culture Techniques