DQA1*0501.TSHR peptides of 183～198、195～210、248～263、301～320、343～362 and 357～376 could bind with high affinity to both HLA-DR3 and HLA-DQA1*0501,all of their IC50 values less than(1 ?mol/L),but they could bind to neither HLA-DR7 nor HLA-DQA1*0201 with high affinity.Conclusion Epitopes of TSHR 183～198,195～210,248～263,301～320,343～362 and 357～376 on the TSHR extracellular domain might be the auto-antigens to cause GD.

Objective To evaluate the clinical significance of determination for serum insulin like growth factor binding protein 3 (IGFBP 3) protease activity. Methods Serum IGFBP 3 protease activity in normal persons, parturient pregnant women, acromegalic patients and patients with growth hormone deficiency (GHD) was determined by Western blot technique. Results Serum IGFBP 3 protease activity in parturient pregnant women, acromegalic patients,normaladultsanduntreatedGHDpatientswere (75.2?2.2)%, (68.6?3.9)%, (22.7?3.1)% and (13.4?4.2)% respectively. Serum IGFBP 3 protease activity in GHD increased to (29.0?6.8)% after treatment with recombining human somatropin (rhGH) for six months. Conclusions Serum IGFBP 3 protease activity is modulated by GH and affects serum levels of members of GH IGF Ⅰ axis.

Objective To review the epidemiologic features of norovirus infection outbreaks in Zhejiang province during 2004-2014.Methods Epidemiological data of norovirus infection outbreaks in Zhejiang province from January 2004 to February 2014 were collected from the Emergency Public Reporting System in Zhejiang Provincial CDC.The distribution of time,area,population,route of transmission and genotype of norovirus were analyzed.Results There were 16 outbreaks of norovirus infections with 2 037 cases during 2004-2014 in Zhejiang province.Eleven outbreaks occurred during February and April,and 13 outbreaks occurred in schools.The outbreaks in schools mainly involved students aged 15-20 years,while other outbreaks took place mainly in the young and middle-aged population.The sex ratio of male to female was 1.05 ∶ 1.Among 16 outbreaks,10 were induced by norovirus G Ⅱ infections,3 were induced by norovirus G Ⅰ infections and 3 were induced by norovirus G Ⅰ and G Ⅱ infections.Fourteen outbreaks were caused by water pollution.Conclusion Outbreak of norovirus infection usually occurs in schools during winter and spring in Zhejiang province,and the epidemic of disease is mainly associated with polluted water.

OBJECTIVETo study the value of serum insulin-like growth factor binding protein-3 (IGFBP-3) levels in differential diagnosis of growth hormone deficiency (GHD).

METHODSTo measure serum IGFBP-3 levels by RIA in normal children and adolescents, GHD children and short-stature children without GHD.

RESULTSSerum level of IGFBP-3 in 129 children with untreated GHD and with no pubertal development was 1.6 +/- 0.9 mg/L, which was less than that in normal group of the same age, but overlapped with the normal children in Tanner stage I. After six-month treatment with recombinant human growth hormone (rhGH), serum level of IGFBP-3 in 59 GHD significantly increased from 1.3 +/- 0.7 mg/L to 2.7 +/- 0.9 mg/L, accompanied by an increase of body heights, growth velocities and serum level of IGF-1. Serum level of IGFBP-3 in 55 short-stature children without GHD was 3.3 +/- 2.2 mg/L, which was not significantly different from that in normal group.

CONCLUSIONSerum IGFBP-3 level can reflect the status of GH secretion in children with GHD and is a useful marker for differential diagnosis of GHD.

Adolescent ; Biomarkers ; blood ; Body Height ; drug effects ; Child ; Diagnosis, Differential ; Female ; Human Growth Hormone ; deficiency ; therapeutic use ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; metabolism ; Male ; Recombinant Proteins ; therapeutic use

Objective To investigate the effect of acute non-isovolemic hemodilution in combination with tranexamic acid on cycle function blood gas and electrolytes with brain tumor surgery. Methods Forty-two patients undergoing brain tumor were randomly divided into two groups. Patients in group A received ANIH plus tranexamic acid , while patients in group B received ANIH alone. Collected blood was transfused before the end of surgery. HR、CVP、MAP,hemoglubin, blood gas and plasma electrolytes were respectively recorded before ANIH(T1), at 0 min (T1) and 1 h (T2) after ANIH, and at the end of operation (T4). Results There were no significant changes in HR, CVP, MAP. At T2, T3, T4, Hb, Hct in both two groups lower than those at T1(P <0.05); at T4, Hb, Hct in group A were higher than those in group B. There were no significant changes in pH , PaO2, PaCO2, BE between the both two groups. There were no significant changes in Na +, Cl-, Ca2+and K+between the both two groups. Conclusion ANIH has little effect on the cycle function and blood gas electrolyte. ANIH in combination with TA has a section blood effect. It can be used in the brain tumor operation with TA security.

Objective To study the effect of acute non-isovolemic hemodilution (ANIH) plus tranexamic acid on bleeding and coagulation factors. Methods Forty-two patients with brain tumor under general anesthesia were randomly divided into group N and group T with 21 patients in each group. Group N was given ANIH , while group T was given tranexamic acid and ANIH. Bleeding, transfusion, urine volume were recorded. Coagulation factors and D-dimer were detected one day before and after surgery. Hemoglobin was recorded before and after ANIH and after auto-blood was transfused. Results There was less bleeding in group T. Hemoglobin in group T was higher after transfusion. No significant difference was found in Group T and group N in terms of urine volume and transfusion rate. Both the two groups had no difference on variation of coagulation factors. Conclusion ANIH with tranexamic acid has no significant effect on coagulation but produces synergetic effect on decreasing bleeding. They can be applied in surgery of brain tumor safely.

Objective To provide a fixtures and feasible injection method for rat tail vein injection.Methods SD rats were randomly divided into A group and B group,thirty rats in each group.Rats in group A fixed by a simple and practical experimental rats fixtures.And rats in group B fixed by common plastic drink bottles.Then the tail vein injection experiment was conducted respectively.Results It took one people 31.2 seconds in group A and 33.1 seconds in group B to finish the experiment from capture to fix rats,and took one people 68.4 seconds in group A to finish the experiment from capture to finish the injection,while it couldn't finish in group B.It took two people 25.4 seconds in group A and 25.8 seconds in group B to finish the experiment from capture to fix rats,and took 63.7 seconds in group A and 85.6 seconds in group B to finish the experiment from capture to finish the injection.Conclusion The experimental rats fixtures can increase the success rate of rats tail vein injection,and shorten the injection time.It is a safe and effective method.

Objective To investigate the effects of controlled hypotension (CH) combined with tranexamic acid (TA) on peri-operative blood loss and coagulation function in patients undergoing brain tumor surgery. Methods Forty patients undergoing brain tumor surgery were randomly allocated into group A and group B with 20 patients in each group. Patients in group A received CH alone, while patients in group B received CH combined with TA. Coagulation factors and d-dimer levels were measured 24 hours before and after surgery. Amount of blood loss, intravenous fluid transfused, urine output and postoperative drainage were recorded. Results D-dimer levels of 24 hours after surgery increased compared with that of 24 hours before surgery. In group B, the d-dimer level increased more than that of group A (P < 0.05). No significant difference was found in coagulation factor levels between group A and group B. Amount of blood loss, intravenous fluid transfused and postoperative drainage flows of patients in group B were lower than that in group A (P < 0.05). There were no significant changes in urine output and fluid infusion volume between two groups. Conclusion CH compared with TA can reduce perioperative blood loss in patients undergoing brain tumor surgery , with no obvious coagulant function abnormality. Collectively, it should be a safe and reliable method for clinical application.

Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).

Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.