Objective To investigate the effect of microcireulation blood flow and altering immunity by Chuanxiongqin,ulinastain and thymosin α1 on sepsis patients.Methods 90 patients were randomly divided into 3 groups(n=30),namely ICU group,Chuanxiongqin group,ulinastain and thymosin α1 group.HLA-DR/CD14+and IL-6,TNF-α,Lac,DD were measured.Results (1)DD showed no significant difference at every time point between ICU group and ulinastain+thymosin α1 group(P＞0.05).DD decreased in Chuanxiongqin group,and was significantly different from the others on the third day.(2)Lac unchanged significantly at every time point in ICU group(P＞0.05).Lac in Chuanxiongqin group and ulinastain+thymeain α1 group tended to decrease,and was statistically different from ICU group on the second day.(3)IL-6 and TNF-α tended to increase at every time point in ICU group(P＜0.05).In ulinastain+thymosin α1 group,IL-6 and TNF-α returned to the level before treatment,HLA-DR/(D14+increased significantly,and was higher than Chuanxiongqin group and ICU group statistically.Conclusion Chuanxiongqin could ameliorate circulation;ulinastsin and thymosin α1 could depress IL-6,TNF-α.So ulinastain and thymosin α1 might protect the immunity of sepsis patients.

Objective To investigate the underlying mechanism of bone marrow mesenchymal stem cells (BMSC) modulating the inflammatory response during the multiple organ dysfunction syndrome (MODS), especially the expression of inflammatory cytokines, which will provide new theoretical and experimental basis of MODS in clinic. Methods BMSC of Sprague-Dawley (SD) rat (female, 4 weeks) was extracted and cultivated, and the 4th passage were used in experimental study. According to the random number table, 60 female SD rats were divided into three groups (n = 20 per group): sham group, MODS group, BMSC group. MODS model in rats was induced by lipopolysaccaride (LPS, 1 mg/kg) via femoral vein injection. Sham group was injected with the sterile phosphate buffer saline (PBS) in the same volume. BMSC group, in which BMSC infusion was started at 2 hours after 0.5 mL LPS stimulation (1×106/cells) through the tail vein. The survival rate was observed after 72 hours in each group. Abdominal aortic blood was collected for routine blood and biochemical examination at 72 hours after operation. Protein microarray was used to detect the related 34 inflammatory cytokines. Signal ratio was defined as the differentially expressed factors when it was more than 2.0 or less than 0.5. And enzyme linked immunosorbent assay (ELISA) was be applied to validate the significant inflammation factor. Meanwhile, the heart, kidney, intestine tissue was harvested, then their pathological changes were observed by hematoxylin eosin (HE) staining.Results 20, 12, 16 rats lived in sham group, MODS group and BMSC group respectively at 72 hours after operation. Compared with the sham group, the indicators (routine blood, liver and kidney function, myocardial enzyme) were apparently unusual, and the heart, kidney, intestine tissue were injured obviously in the MODS group. After BMSC administration, the organ function was improved and tissue damaged was alleviated significantly. Protein microarray showed that interleukin-4 (IL-4) and receptor for advanced glycation end products (RAGE) were significantly different in 34 goal cytokines. The signal ratio change of IL-4 was 0.397, 1.124, 2.826 respectively, and the signal ratio of RAGE was 6.197, 1.552, 0.250, respectively in MODS/sham group, BMSC/sham group, BMSC/MODS group. ELISA validated the result that the expression level of IL-4 decreased significantly (ng/L:3.59±1.21 vs. 29.10±5.78) and the expression level of RAGE increased significantly (ng/L: 1.09±0.04 vs. 0.11±0.03) in MODS group as compared with sham group (bothP < 0.05). Compared with the MODS group, the level of IL-4 was obviously higher than that in BMSC group (ng/L: 9.59±2.21 vs. 3.59±1.21,P < 0.01), and RAGE decreased significantly (ng/L: 0.29±0.07 vs. 1.09±0.04,P < 0.05).Conclusions BMSC administration can regulate the expression of IL-4 and RAGE in the rats subjected to MODS. Moreover, BMSC can promote the restoration of tissue and organ function, thus improve the survival rate. BMSC may be the target in cell therapy for the inflammatory disease.

To establish a quantitative assay for telomerase activity and analyze the telomerase activity in peripheral blood mononuclear cells (PBMNC) from patients with acute leukemia, a fluorescent dye, PicoGreen, was added to the products after telomere repeat amplification protocol. The samples were excited at 480 nm and the fluorescence emission intensity was measured at 520 nm using a spectrofluorometer. Telomerase activity was detected in PBMNCs from 20 cases of normal individuals and 25 patients with acute leukemia. The results showed that the fluorescence of PicoGreen binding to double-stranded DNA specifically was enhanced with increase of DNA quantities. In conclusion, the met hod is rapid, simple and quantitative, the telomerase activities of PBMNCs from acute leukemia patients are significantly higher than that of the normal controls.
Acute Disease ; Adolescent ; Adult ; Aged ; Cell Line ; DNA, Neoplasm ; genetics ; metabolism ; Female ; Fluorescent Dyes ; chemistry ; Humans ; Leukemia ; blood ; enzymology ; genetics ; Leukocytes, Mononuclear ; enzymology ; Male ; Middle Aged ; Organic Chemicals ; Telomerase ; chemistry ; genetics ; metabolism

Objective To study the changes of serum TNF-α and NO in neonatal rats with endotoxic shock and its relationship with myocardial cells damge and to explore the protection effect of dexamethasone (DXM) on neonatal rats with endotoxic shock. Methods Nine of the 117 seven-day-old healthy neonatal Wistar rats were used as the pre-experimental base value control group, and the other 108 rats were randomly divided into control group, endotoxic shock group (LPS group, LPS 5 mg/kg) and treatment group (DXM group, LPS 5 mg/kg+DXM 5 mg/kg). Before and after injection of LPS (2, 4, 6 and 24 hs), 9 rats in each group were sacrificed and blood samples were collected to detect TNF-α by ELISA and NO by nitrate reductase. Myocardial super-microstructure was observed under an electron microscope. Results ① In the LPS group, the concentration of serum TNF-α peaked at 2 h, and it decreased to the level of control group after 6 hs. In the DXM group, the level of TNF-α at 2 h was higher than that of control group (P<0.05), but lower than that of the LPS group (P<0.05). At 4 hs the level TNF-α in DXM group was lower than that of the LPS group (P<0.05), but was not different than that of the control group (P>0.05). From 6 h after injection, the differences of TNF-α levels in 3 groups were not obvious (P>0.05). ② In the LPS group, the concentration of NO rose after 2 h (P<0.01), and peaked at 24 h (P<0.01). After 2 hs, the levels of NO in the DXM group were lower than those of the LPS group (P<0.01). ③ Six hours after the injection of LPS, a little mitochondria of myocardial cells in the LPS group appeared to develop vacuolae-like degeneration. At 24 h, most mitochondria of myocardial cells in the LPS group presententecl with vaculea-like degeneration and necrosis. The myocardial fibers were broken. While in the DXM group, the changes in the super-microstructure at 24 h were not as serious as those which took place in the LPS group. Conclusions TNF-α and No were involved in the damage of myocardial cells in neonatal rats with endotoxic shock. DXM could partly protect the myocardial cells from damage by inhibiting the production of TNF-α and NO.

OBJECTIVETo explore the norms of treatment of acute organophosphorus pesticide poisoning (AOPP), and observe the curative effect.

METHODSOn basis of the pre-research, the norms of treatment of AOPP were summarized, and a multi-center clinical trial was performed in 6 hospitals selected from high incidence of AOPP in Shandong Province.

RESULTS422 patients of AOPP in 6 hospitals in observation period were treated and observed by the norms of treatment. Among them, the proportion of oral poisoning was 97.16%, middle and severe degree were 87.44%. Compared with themselves 2 years ago before standard treatment, the curative effect of the norms of treatment for AOPP was much better than before. The mortality rate of AOPP declined from 9.87% to 1.66% (Chi2 = 27.92, P < 0.01), that was much better than the average therapeutic effect level of all our province in the same period (the mortality rate: 8.92%) (Chi2 = 26.05, P < 0.01). The average amount of atropine [(37.54 +/- 17.76) mg], dropped greatly [(1280.70 +/- 69.22) mg] (U = 439.22, P < 0.01).The usage of atropine by continuous intravenous injection with venous pump was better than ordinary intravenous injection. The mean dosage of pralidoxime chloride increased twice than the previous (U = 19.48, P < 0.01). There was no drug poisoning.

CONCLUSIONThe standard treatment of AOPP is urgently needed in our country, especially in rural area. By this trial, the satisfactory effect of the norms of treatment for AOPP summarized is observed and it reduces the fatality rate remarkably.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Organophosphate Poisoning ; Pesticides ; poisoning ; Poisoning ; therapy ; Standard of Care ; standards ; Young Adult

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.
Apoptosis Regulatory Proteins ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; Transfection ; U937 Cells

OBJECTIVETo sum up the experience in the treatment of obstructive azoospermia by intracytoplasmic sperm injection (ICSI).

METHODSWe retrospectively analyzed 107 cases of obstructive azoospermia treated by ICSI in our center from Jan. 2006 to Dec. 2008, and compared the rates of fertilization, cleavage and pregnancy between the patients with congenital bilateral absence of vas deferens (CBAVD) and those with non-CBAVD.

RESULTSA total of 949 oocytes were injected for the 107 patients undergoing ICSI, of which 678 (71.4%) were fertilized and 605 (89.2%) cleaved, with 44 pregnancies (41.4%). Of the 442 oocytes injected for the 49 patients with CBAVD, 308 (69.6%) were fertilized and 279 (90.6%) cleaved, with 27 pregnancies (55.1%), and of the 507 oocytes injected for the 58 cases induced by inflammation or surgery, 370 (72.9%) were fertilized and 326 (88.1%) cleaved, with 17 pregnancies (29.3%). The rate of pregnancy was significantly higher in the CBAVD than in the non-CBAVD group (P < 0.01), but there were no significant differences in the rates of fertilization and cleavage between the two groups (P > 0.05).

CONCLUSIONPESA or TESE combined with ICSI is an effective approach to the treatment of male infertility induced by obstructive azoospermia, which may achieve a higher rate of pregnancy in patients with CBAVD than in those with non-CBAVD. Inflammation or surgery may not only cause the obstruction of the deferent duct, but also affect sperm quality, and consequently reduce the potentiality of embryonic development.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Treatment Outcome

OBJECTIVETo explore the difference on the prevalence and the risk factors of type 2 diabetes(T2DM) between Yi farmer and immigrants in Liangshan, Sichuan province.

METHODSA representative sample of 2878 Yi people (including 1549 farmers and 1329 immigrants) aged from 16 to 76 was selected by the method of multistage and cluster sampling in Liangshan, Sichuan province, China, during 2007 - 2008. The samples were divided into 5 groups by the factor of age (16-, 25-, 35-, 45- and 55-76). The standardized prevalence of T2DM was calculated by the national census data in 2000. Logistic regression analysis was used to study the related risk factors of T2DM.

RESULTSThe prevalence of Yi farmer was 4.33% (67/1549) (male: 6.15% (42/683), female: 2.89% (25/866)), and that of Yi migrants was 9.03% (120/1329) (male: 11.31% (88/778), female: 5.81% (32/551)). The standardized prevalence (SP) was calculated by the data of national census 2000. The SP of Yi farmer was 5.97%in male, and that of the female was 2.40%. The SP of Yi migrant was 10.25% in male, and that of the female was 6.29%. For Yi people, sex (male versus female, OR = 1.69, 95%CI: 1.02 - 2.81), age (versus the group aged 16- and 25-, group aged 35 to 54: OR = 5.04, 95%CI: 2.93 - 8.69; group aged above 54: OR = 6.19, 95%CI: 3.23 - 11.86), hypertension (versus normal group, borderline hypertension value: OR = 1.61, 95%CI: 1.08 - 2.38; hypertension group: OR = 2.40, 95%CI: 1.37 - 4.22), smoking (OR = 1.55, 95%CI: 1.01 - 2.37), triglyceride (TG) level (OR = 1.65, 95%CI: 1.10 - 2.46) and high-density lipoprotein cholesterol (HDL-C) level (OR = 1.64, 95%CI: 1.13 - 2.37) were the positive factors correlated with T2DM, and drinking (the alcohol volume from 22.67 to 52.50 g/d ) was negative factor (OR = 0.53, 95%CI: 0.30 - 0.95) correlated with T2DM.

CONCLUSIONThe prevalence of T2DM in Yi immigrants was higher than that in Yi farmers;sex, age, blood pressure, smoking, TG, HDL-C, drinking were influencing factors of T2DM.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; Ethnic Groups ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Rural Population ; Young Adult

Objective:To study diagnostic value of left atrial diameter(LAD),lipoprotein(a)[LP(a)]combined D-dimer(D-D)for left atrial thrombus(LATH)in patients with atrial fibrillation(AF).Methods:According to results of transesophageal echocardiography,a total of 367 AF patients treated in our hospital were divided into LATH group(n=67)and no LATH group(n=300).General clinical data,LAD,serum Lp(a)level and positive D-D rate were compared between two groups,and diagnostic value of above indexes for LATH in AF was ana-lyzed.Results:Compared with no LATH group,there were significant rise in LAD[(40.93±4.69)mm vs.(44.65±4.31)mm],serum Lp(a)level[(17.05±2.31)mg/dl vs.(27.42±3.06)mg/dl]and positive D-D rate(5.67%vs.22.34%)in LATH group(P=0.001 all).ROC curve indicated that AUC of Lp(a),positive D-D,LAD single detection and triple combined detection diagnosing LATH in AF was 0.711,0.584,0.728 and 0.801 respectively;sensitivity was 43.28%,22.39%,64.18%and 65.67%respectively;and specificity was 92.00%,94.33%,76.67%and 80.67%respectively.AUC of triple combined detection was significantly higher than that of any single detection,and sensitivity and specificity were significantly higher than those of Lp(a)and positive D-D single detection(P＜0.05 or＜0.01).Conclusion:Combined detection of Lp(a),positive D-D and LAD possess high diagnostic value for left atrial thrombus in atrial fibrillation.