AIM: To explore the variation of blood biochemistry and arterial blood gas of patients with systemic inflammatory response syndrome (SIRS) in the early time after trauma and improve the diagnosis and first aid. METHODS: Eighty-eight patients with trauma from August 2003 to February 2004 were divided into two groups by their AIS-ISS90 score. The data of temperature, pulse, respiratory rate, white blood cell counts, Hb, blood glucose and arterial blood gas (PaO 2, PaCO 2, HCO 3 -, AG) were collected and compared with each group by statistic methods. RESULTS: Of the 88 patients, 49 underwent SIRS, 12 in light trauma group (ISS≥16) and 37 in severe trauma group (ISS45 mmHg or 0 05). 13 patients had MODS in severe trauma group and 2 died while none had MODS or died in light trauma group. CONCLUSION: Application of AIS-ISS90 and SIRS-related blood biochemistry and arterial blood gas is beneficial for the diagnosis and treatment for patients in the early time after trauma. [

Child ; Humans ; Immunoglobulin A ; blood ; Immunoproliferative Small Intestinal Disease ; diagnosis ; drug therapy ; etiology ; Male

BACKGROUND: Leptin is a kind of polypeptide hormone secreted by fatty tissue, previous studies have shown that leptin plays a certain role in the formation of atherosclerosis.OBJECTIVE: To investigate the effects of leptin on the expression of tumor necrosis factor-alpha (TNF-α) in RAW264.7 cells, and investigate the possible mechanism from the angle of the change of nuclear factor-κB (NF-κB) activity.DESIGN: A controlled observational experiment.SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiments were carried out in the Department of Biochemistry and Molecular Biology, Tongji Medical College, and the Department of General Surgery, Union Hospital affiliated to Huazhong University of Science and Technology between April 2005 and February 2006.The cultured RAW264.7 cells were divided into leptin treated groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. Each group had 3 sub-wells,and the experiments were repeated for 3 times.METHODS: Mouse macrophage RAW264.7 cells were incubated into a 6-well plate at the density of 109 cells L-1, cultured in RPMI-1640 culture medium containing bovine serum of 0.1 in volume fraction. When the cells grew to 80%, serum-free culture medium Opti-MEM was changed to culture for another 24 hours, and then the cells were divided into leptin groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. After the cells were incubated with leptin for 4 hours, the expression of TNF-α mRNA expression in RAW264.7 cells was detected with reverse transcription-polymerase chain reaction (RT-PCR). After the RAW264.7 cells were incubated with leptin for 1, 3, 6 and 9 hours, the expression of TNF-c protein expression was detected with double antibody sandwich enzyme-linked immunosorbent assay (ELISA). After the RAW264.7 cells were incubated with leptin for different times, the activity of NF-κB was detected analyzed with electrophoretic mobility shift assay. Another, the RAW264.7 cells were treated with or without 50 μmol/L leptin and/or 100 μmol/L PS1145 (I kappa B kinase specific inhibitor)divided into four groups: blank control group, I kappa B kinase specific inhibitor PS1145 (10 μmol/L) treated group, leptin (50 μmol/L) treated group, leptin (50 μmol/L) + PS1145 (10 μmol/L) group. Aftere incubated for 6 hours, the activity of NF-κB and expression TNF-α mRNA were detected respectively.MAIN OUTCOME MEASURES: ① Effect of leptin of different concentration on the expression of TNF-α mRNA and protein in RAW264.7cells; ② Effect of leptin of different concentration on the activity of NF-κB in RAW264.7 cells; ③ Influence of inhibition I kappa B kinase activity inhibition on expression of TNF-a induced by leptin in RAW264.7cells.RESULTS: ① After the RAW264.7 cells were treated with leptin of different concentration, the TNF-α mRNA level was elevated in a dose-dependent manner, and it reached the peak value emerged in the 50 μg/L leptin treated group. ② The expression of TNF-α protein increased in dose-dependent and time-dependent manners, and it reached the peak val ue at 6 hours in the 50 μg/L leptin treated group. ③ The activity of NF-κB was also positively correlated with the leptin concentration, and it was the highest value at 6 hours treated with 50 μg/L leptin (P ＜ 0.05). ④ I kappa B kinase activity inhibition only partially suppressed the leptin induced elevation of TNF-α expression induced by leptin.CONCLUSION: Leptin can increase the expression and secretion of TNF-α in RAW264.7 cells directly in both dose-dependent and time-dependent manners, and the mechanism may be correlated with the activated NF-κB induced by leptin. It may be one of the mechanisms of atherosclerosis induced by leptin.

AIM:To set up a method for determining epimedin C and icariin in Xianling Guobao Capsules(Herba Epimedii,Radix et Rhizoma Salviae miltiorrhizae,Fructus Psoraleae,Radix Rehmanniae,etc.). METHODS:The chromatographic conditions included the column of Spherisorb C 18 (4.6 mm?250 mm,5 ?m),the mobile phase was acetonitrice and water as gradient eluent was at a flow rate of 1.0 mL/min,the detection wavelength was set at 270 nm and the column temperature was at 25 ℃. RESULTS:The linear range of epimedin C was 0.22-2.20 ?g and icariin was 0.04-0.40 ?g,respectively. The average recovery of epimedin C and icariin were 103.2% (RSD=3.1%) and 97.8% (RSD=3.2%),respectively. CONCLUSION:The method is reliable,stable and well reproducible,and can control the quality of Xianling Guobao Capsules.

Objective To analyze the characteristics and the related risk factors of the contractility of pelvic floor muscles in old women with pelvic floor dysfunction(PFD).Methods A cohort of 125 elderly patients with symptoms of pelvic floor dysfunction,including 60 of postpartum urinary incontinence(SUI)and 65 of pelvic organ prolapse(POP),were recruited.60 women volunteers with normal pelvic floor function were recruited as a control group.A questionnaire survey was conducted to record the general situation of the subjects.The pelvic floor muscle strength was determined by the methods of electromyography and pressure feedback recording.Results The body mass index,the average number of delivery,newborn baby's body weight were significantly higher in the SUI and POP group than in control group,with statistically significant difference(F=5.29,6.27,5.26,P=0.007,0.003,0.008),but without statistically significant difference between SUI and POP groups (P =0.674,0.554,0.578).The rapid contractility value was significant lower in SUI group than in POP and control groups(all P＜0.01),and the sustained contractility value was significant lower in POP group than in SUI and control groups(all P＜0.01).Conclusions The postpartum pelvic floor dysfunction may be correlated with BMI,vaginal delivery rate,the average number of delivery,newborn baby's body weight and a history of increased abdominal pressure.The postpartum pelvic floor dysfunction may be associated with the declined contractility of pelvic floor muscle,among which the POP may be associated with class Ⅰ muscle and the SUI with class Ⅱ muscle.

Objective To determine levels of nuclear factor (NF)-κB, interleukin (IL)-10, and visfatin in adipocytes treated by different degrees of intermittent hypoxia (IH), and to investigate the mechanism of IH leading to insulin resistance (IR). Methods The cell model of intermittent hypoxia/re-oxygenation (IH/ROX) in obstructive sleep apnea (OSA) was established. Differentiation mature 3T3-L1 adipocytes, were randomly divided into 10 groups including four different-frequency intermittent hypoxia groups(IH1-4, fixed intermittent hypoxia scheme for 1.5%O2 45 s and then re-oxygen 21%O2 for 2 min 15 s, 4 min 15 s, 5 min 45 s and 8 min 45 s, 60 times circulation), and their normal oxygen control groups (SC1-4, instead each IH group 1.5%O2 to 21%O2, the rest groups were treated as same as IH group), continuous hypoxia group (CH, 10%O2 for 6 h) and normal oxygen control group (CC, 21%O2 for 6 h). ELISA method was used to determine the levels of IL-10 and visfatin in the supematant of adipocytes. Western blot method was used to determine the protein levels of NF-κB p65 and visfatin. Real-time PCR method was used to determine the mRNA levels of IL-10 and visfatin. Results The protein and mRNA expressions of IL-10 were significantly lower in IH group and CH group than those of control groups (P<0.01). The levels of NF-κB p65 protein were significantly increased in IH group and CH group than those of control group. The protein and mRNA expressions of visfatin were significantly higher in IH1, IH2 and CH groups than those of control group (P<0.01). Conclusion As a prominent feature of OSA pathophysiology, IH may take part in insulin resistance of OSA patients by abnormally secreting NF-κB, IL-10 and visfatin in adipocytes.

Objective To investigate the related factors of early liver cancer recurrence associated with elevated fasting glucose levels after radical resection of liver cancer.Methods The retrospective cohort study was adopted.The clinical data of 145 patients with liver cancer who were admitted to the Nanfang Hospital of Southern Medical University from October 2009 to June 2013 werc collected.After radical resection of liver cancer, 111 patients with average level of fasting blood glucose (FBG) ＜ 6.1 mmol/L and 34 patients with level of FBG ≥ 6.1 mmol/L were divided into the normal group and the hyperglycemic group, respectively.The blood glucose levels of patients during the period of hospital stay and follow-up were observed.The status of early recurrence and risk factors affecting recurrence in the 2 groups were analyzed.Follow-up of outpatient examination was applied to patients up to July 1, 2015 with the first recurrence as the end point.Patients had reexaminations once every month within postoperative month 6 and once every 3 months after 6 months.The general information [gender, age, FBG, underlying hepatopathy, preoperative AFP, alanine transaminase (ALT), aspartate aminotransferase (AST), Child-Pugh stage, Barcelona Clinic Liver Cancer (BCLC) stage, history of alcohol drinking], surgical data (surgical method, operation time, number of blood transfusion, volume of intraoperative blood loss), pathological data (histopathological differentiation, number of tumor nodules, diameter of maximal tumor, liver cirrhosis) and follow-up data (postoperative AFP, imaging findings, recurrence time, preventive chemotherapy) were collected.Measurement data with normal distribution were presented as-x ± s and analyzed by the t test.Measurement data with skewed distribution were presented as M (range) and analyzed by the non-parametric test.Count data were analyzed using the chi-square test.With the first recurrence as the end point, the tumor-free survival rate was drawn using the Kaplan-Meier method, and the comparison was analyzed by the logrank test.The risk factors affecting recurrence were analyzed using the Kaplan-Meier method for proportional hazards assumption test firstly, and then eligible factors were done using Log-rank test in the univariate analysis.The multivariate analysis was done using the COX model.Results The postoperative 1-, 2-year overall recurrence rates of liver cancer were 28.3％ (41/145) and 45.5％ (66/145).And 1-, 2-year recurrence rates of liver cancer were 21.6％ (24/111) and 36.9％ (41/111) in the normal group and 50.0％ (17/34) and 73.5％ (25/34) in the hyperglycemic group, respectively, showing significant differences between the 2 groups (x2=10.335, 14.053, P ＜ 0.05).The univariate analysis showed that FBG, Child-Pugh stage, volume of intraoperative blood transfusion and postoperative AFP were risk factors affecting tumor-free survival rate after radical resection of liver cancer (x2 =17.591,6.492, 10.690,12.820, P ＜ 0.05).The tumor-free survival rates at postoperative month 24 in the normal group and hyperglycemic group were 63.1％ and 26.5％ respectively, showing significant difference between the 2 groups (x2=17.591, P ＜ 0.05).The results of multivariate analysis showed that level of FBG ≥6.1 mmol/L, volume of intraoperative blood transfusion ＞ 200 mL and postoperative level of AFP ＞ 8.1 pg/L were independent risk factors affecting tumor-free survival rate after radical resection of liver cancer (RR =2.542, 2.028, 2.724, 95 ％ condifence interval : 1.529-4.225,1.183-3.479,1.635-4.538, P ＜ 0.05).Conclusions Elevated FBG level has a stimulative effect on early recurrence of tumor after radical resection of liver cancer.As a result, monitoring and controlling of blood glucose after operation is helpful in decreasing the early recurrence rate of patients with liver cancer.

Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.

Objective:To study the curative efficacy of decitabine combine with CAG regimen in the treatment of acute myeloid leukemia and its effects on the serum Interferon-γ (IFN-γ),alpha hydroxybutyrate dehydrogenase (HBDH) and lactate dehydrogenase (LDH) levels.Methods:70 cases of patients with acute myeloid leukemia who were treated from February 2013 to October 2016 in our hospital were selected as research objects.According to the random number table,the patients were divided into the observation group (n=35) and the control group (n=35).Both groups of patients were treated conventional treatment.The control group was treated with CAG scheme,intravenous injection of aclarubicin,20 mg each time,granulocyte colony stimulating factor,300 μg each time,subcutaneous intravenous cytarabine,10～15 mg/m2 each time,while the observation group was treated with intravenous drip of decitabine on the basis of control group,15 mg/m2 each time.Then the therapeutic effect,serum interferon-γ (IFN-γ),alpha hydroxybutyrate dehydrogenase (HBDH),lactate dehydrogenase (LDH) levels before and after treatment,incidence of adverse reactions were compared between two groups.Results:After treatment,the total effective of observation group was significantly higher than that of the control group [91.42％(32/35)vs68.57％(24/35)] (P ＜0.05);the serum IFN-γ,HBDH,LDH levels were significantly higher than those of the control group [(3.21± 1.01)pg/ml vs.(5.13 ± 1.90)pg/mL,(103.62± 26.39)U/L vs.(118.80± 28.60)U/L,(101.36± 27.32)U/L vs.(123.08 ± 30.59)U/L] (P ＜0.05);the incidence rate of adverse reactions was significantly lower than that of the control group [20.00％(7/35) vs.42.85％(15/35)(P ＜0.05)].Conclusion:Decitabine combined with CAG regimen was more effective for acute myeloid leukemia than CAG regimen alone,which might be related to reduce the serum levels of IFN-γ, HBDH and LDH.

Objective To study the possible relationships between expression of matrix metalloproteinase(MMP)2,9 and the pathogenesis of preeclampsia in which trophoblast invasion is impaired. Methods MMP-2,9 expression were detected by immunohistochemistry streptavidin-biotin complex (SABC)method in 20 normal term placentae and 20 preeclampsia placentae,respectively.In addition, mRNAs for MMP-2,9 were analyzed by real time PCR in both groups.Results The intensities of both MMP-2 and MMP-9 immunostaining in preeclampsia placentae were significantly declined compared to those of normal term placentae(P