Objective To study two new factor Ⅸ mutations Cys82Ser and Ile288Ser in vitro and research the molecular mechanism of haemophilia B. Methods PcDNA3. 1 ( - ) FⅨwt expression plasmid was prepared. The mutated FⅨcDNA expression plasmids, PcDNA3.1 ( - ) FⅨM1 (Cys82Ser) and PcDNA3. 1 ( - ) F Ⅸ M2 (Ile288Ser) were constructed by megaprimer method respectively. Transient expression experiments were performed using HEK293 cells transfected with the expression vectors containing the wild-type or the mutation recombinant cDNA. PcDNA3. 1 ( - ) was used as a blank control. The expression proteins were detected by ELISA, factor activity assay and flourescence stain. Results The results suggested that the two FⅨ gene mutations did not induce the reduction of the mutant FⅨ mRNA compared with the wild-type FⅨ mRNA. The FⅨ:Ag in culture media and cell lysate of wild type conduct were assigned as 100. 0%. The results of PcDNA3.1 ( - ) FⅨ M1 mutation protein were (27. 1 ± 5. 2)% and (99.4 ±4. 1)% respectively. For PcDNA3. 1( - )FⅨM2, the results were (5.3 ± 1.8)% and (31.7 ±2. 5)% respectively. The FⅨ: C in culture media of wild type conduct was also assigned as 100. 0%. Then the two types of mutant protein were ( 8. 5 ± 3.2 ) % and ＜ 1%, respectively. Immunofluorescence microscopy result suggested that the intensity of perinuclear spot was reduced in cells transfected with PcDNA3.1 ( - ) FⅨM2 while staining for PcDNA3. 1 ( - ) FⅨM1 was predominantely diffuse without perinnclear enhancement. Conclusions These results strongly suggest that the FⅨ Cys82Ser mutation protein is not been correctly folded, by any possibility. The mutation protein has secretion defect. The secretion dysfunction and the protein degradation intracellular are possiblely the molecular pathology of Ile288Ser mutant protein.

Objective To evaluate the role of 18F fluorodeoxyglucose positron emission tomography computed tomography (PET-CT) in nasopharyngeal carcinoma (NPC) after radiotherapy. Methods A total of 27 NPC patients received 18FDG PET-CT 8-32 weeks after radiotherapy. All the patients were followed up for about 12 months after the examination. Metastasis and residual were evaluated by PET-CT. The correlation between SUV and prognosis was analyzed. Results Of these 27 patients, metastasis was found in 2 patients by PET-CT. Local persistence was diagnosed as for SUV≥2. 5 by PET-CT in 20 patients, among whom 18 were confirmed by biopsy and then received brachytherapy or conformal radiotherapy. One year local control and survival rates were 70% and 81%. Based on SUV, the patients were divided into group one for SUV between 2. 5 and 5(9 patients) or group two for SUV≥5 (11 patients). In group one and group two, the one year local control rate, survival rate and metastasis rate were 67% , 55% (P=0.670) , 64% ,89%(P=0.319), and 22% , 82% (P =0. 022) , respectively. Conclusions PET-CT is valuable for the identification of residual nasopharyngeal carcinoma. SUV of residual tumor is related to metastasis.

Objective To study the drug resistance of Staphylococcus aureus in 2006 -2011 ,and to provide the evidence for treatment infection .Methods The isolated bacteria were identified and antibiotic sensitivity were tested by automated system in 503 Staphylococcus aureus collected from 2006 to 2011 .Methicillin resistant Staphylococcus aureus(MRSA) was screened by oxacillin disk diffusion .Results 503 strains mainly derived from secretion and sputum .The incidence of MRSA was 44 .9% during 6 years . Detection rate of MRSA was decreased year by year .The difference was statistically significant between 2009 ,2010 ,2011 and 2006 , 2007 ,2008(P<0 .05) .No resistance to quinupristin/dalfopristin ,linezoiid ,vancomycin and nitrofurantoin was found .The resistance of Staphylococcus aureus was below 30% to levofloxacin ,imipenem ,compound sulfamethoxazole and rifampicin ,above 80% to ce-fazolin and penicillin .Although the resistant to cefazolin ,levofloxacin ,imipenem was risen ,the resistant rate of rest antibiotics was downed year after year .Conclusion Monitoring of drug resistance should be strengthened .The antimicrobial therapy should be de-fined on the basis of drug-sensitive test in order to control the incidence of infection and to delay the growth of clinical resistant strains of Staphylococcus aureus .

The aim of this study is to evaluate the application of untrasound-guided local anesthesia in alleviation of pain after percutaneous liver biopsy.The clinical results of 1417 cases of percutuneous liver biopsy were retrospectively analyzed.In 896 patients under ultrasound-guided local anesthesia 51 felt pain after liver biopsy,while in 521 patients whose local anesthesia without ultrasound guidance,143 felt pain (5.7% vs.27.4%,X~2=118.63,P＜0.01).The results indicate ultrasound-guided local anesthesia can effectively alleviate pain after percutaneous liver biopsy.

Objective To understand the diagnostic value of blink reflex (BR) in central neuropathy by observing the variations of blink reflex in diabetic patients. Methods Electrical pulse was used to stimulate the supraorbital nerve at the supraorbital notch. The responses of bilateral orbicular muscles of the eye were recorded, the variations of the latency and amplitude of R 1 and R 2 recorded on the side of stimulation and of R′ 2 recorded on the contralateral side were measured. Results There was significant prolongation in the latency and lowering of the amplitude of R 1, R 2 and R′ 2 in the diabetic patients compared with the control group. The abnormality rate of BR increased gradually with the prolongation of diabetic duration. Conclusion BR examination might provide an objective index for nervous damage in diabetic patients and it is helpful in early diagnosis of diabetic neuropathy.

Objective To study the immune regulation of antisense peptide in rats by observing immune function of activity fragments of thyrotropin receptor (TSHR) and their corresponding antisense peptides. Methods TSHR peptides TR1, TR2, TR3 and their antisense peptides RT1, RT2, RT3, and three pairs of complementary peptides were injected into rats of different groups respectively, and the serum levels of TT_3, TT_4, TSHR antibody (TRAb), thyroid stimmulating antibody, thyroid blocking antibody and TSH antibody (TSHAb) and pathological changes in thyroid tissue were investigated. Results Serum TRAb could be induced when each of three fragments of TSHR was injected into rats; TRAb and TSHAb were induced by RT1 or RT2; epithelial hyperplasia and lymphocytic infiltration observed in thyroid tissue of rats injected with TR2 could be abated by injecting RT2 subsequently. Conclusion The results suggest that all 3 TSHR fragments are shown to be immunogenic and are capable to induce TRAb; both RT1 and RT2 show their effect on immune regulation and are idiotypic of TSHR peptides; On the other hand, the humoural and cell immunities are ameliarated by injection of antisense peptides. Therefore, it is possible that antisense peptides may be involved in immune regulation via immune network.

OBJECTIVE:To establish a method for the content determination of polymer in Cefotaxime sodium and tazobactam sodium(6∶1)for injection. METHODS:High performance size exclusion chromatography(HPSEC)was performed on the column of TSK-GEL G2000SWXL with mobile phase of 0.01 mol/L phosphate buffer (pH 7.0) at a flow rate of 1.0 ml/min,detection wavelength was 254 nm,injection volume was 10 μl and the column temperature was 25 ℃. RESULTS:Cefotaxime and the poly-mers were well-separated;the linear range of cefotaxime was 2.3-226.4 ng(r=0.999 9);RSDs of precision and reproducibility tests were no more than 0.79%;results of durability test showed the changes of column temperature,flow rate,wavelength,pH and mobile phase salt concentration had little effects on the separation among different polymer peaks and between polymer peaks and main peaks. CONCLUSIONS:The method is simple,specific and sensitive,and can be used for the content determination of poly-mer in Cefotaxime sodium and tazobactam sodium(6∶1)for injuction.

Objective To evaluate the effectiveness of a newly designed modular flexible ureteroscope with holmium laser lithotripsy for the treatment of renal calculi based. Methods 40 patients were treated with modular flexible ureteroscopic lithotripsy (German Polydiagnost) with holmium laser. 40 patients were treated by traditional flexible ureteroscopic lithotripsy. Their therapeutic effects were compared. Results For modular flexible ureteroscopic lithotripsy group, 37 patients underwent successful operation, with lithotripsy with operation time of (86.0 ± 34.4) min and postoperative hospital stay of (5.3 ± 1.6) days. No severe complication including ureteral perforation, high fever or severe bleeding occurred. One month after the operation, KUB and B-ultrasonography showed complete stone free in 35 patients. Residual calculi (0. 4 cm in diameter) in the calyces were found in 2 patients. Medication and postural drainage therapy were applied. Two weeks later , KUB and B-ultrasonography showed that the residual stones have been removed completely. There was no significant differences in stone size , operation time, rate of calculus clearance, the incidence of postoperative complications, operating time and duration of postoperative hospitalization between the traditional flexible ureteroscopic lithotripsy group and modular ureteroscopic lithotripsy group. Conclusions The modular flexible ureteroscope is effective and safe in treating reanl calculi. It has similar surgical efficacy as traditional flexible ureteroscope , but is more costly effective in terms of maintenance costs.

AIM: The selective recognition of the sense peptides which are located in special regions of thyrotropin receptor (TSHR) by their corresponding antisense peptides has been investigated. Three pairs of sense and antisense peptides were named TR1 (aa37-45) and RT1 (aa45-37), TR2 (aa353-366) and RT2 (aa366-353), TR3 (aa648-655) and RT3 (aa655-648). METHODS: To prepare three affinity chromatography columns, antisense peptides were immobilized, called RT1-sepharose 4B, RT2-sepharose 4B and RT3-sepharose 4B, respectively and investigate the retardative behavior for each of native peptide TR1, TR2 or TR3 on above columns with stepwise elution. RESULTS: Each of the three immobilized antisense peptides recognized and retarded its corresponding sense peptide-TR1, TR2 or TR3 instead of those non-complementary peptides. Immobilized RT1 recognized free TSHR protein molecule as well. In additional, bovine thyrotropin was recognized by immobilized TR1. CONCLUSION: The results indicate that molecular recognition theory exsits in thyrotropin receptor system. It may be useful to isolate biological molecules and to locate epitopes of TSH on TSHR molecule. Otherwise, antisense peptide may be used for treatment of experimental autoimmunolized thyroid disease (AITD) in the rat. [

Objective To determine the role of activated status of peroxisome proliferator-activated receptorγ/nuclear factor-κB ( PPAR-γ/NF-κB ) in coagulation disorders induced by sepsis. Methods Forty male Sprague-Dawley ( SD ) rats were randomly divided into four groups, n = 10 in each group: control group, lipopolysaccharide ( LPS ) challenged group, rosiglitazone ( ROSI, selective agonist of PPAR-γ) pretreatment group, and GW9662 ( PPAR-γ antagonist ) pretreatment group. The sepsis model was reproduced by injection of 6 mg/kg LPS via sublingual vein, and the rats in control group were injected with 2 mL/kg normal saline. The rats in ROSI pretreatment group were given 0.3 mg/kg ROSI by sublingual venous injection followed by injection of LPS 30 minutes later;and in GW9662 pretreatment group rats were given 0.3 mg/kg GW9662 by sublingual venous injection followed by 0.3 mg/kg ROSI 15 minutes later, followed by injection of LPS 30 minutes later. Blood was collected at 4 hours after LPS administration, and the expressions of PPAR-γ and NF-κBp65 in peripheral blood mononuclear cell ( PBMC ) were determined with immunocytocheminal technique and graph analysis. Plasma prothrombin time ( PT ), activated partial thromboplastin time ( APTT ), fibrinogen ( FIB ), and D-dimer were determined simultaneously. Results① PPAR-γ/NF-κB pathway: the expressions of PPAR-γ and NF-κBp65 were lowered in control group, and they were expressed in cytoplasm. In LPS challenged group the expression of PPAR-γ ( gray value ) was slightly increased but with no significant difference as compared with control group ( 111.01±4.06 vs. 98.46±5.99, P >0.05 ). In ROSI pretreatment group the expression of PPAR-γ( gray value ) was significantly higher than that in LPS challenged group ( 214.38±5.79 vs. 111.01±4.06, P<0.01 ), with dislocation into nuclei. In GW9662 pretreatment group the expression of PPAR-γ ( gray value ) was lowered but without significant difference compared with that of control group ( 44.21±2.64 vs. 98.46±5.99, P>0.05 ). In LPS challenged group the expression of NF-κBp65 ( gray value ) was significantly higher than that in control group ( 249.48±6.86 vs. 105.81±10.19, P < 0.01 ), and it was translocated into the nuclei. In ROSI pretreatment group the expression of NF-κBp65 ( gray value ) was significantly lower than that in LPS challenged group ( 102.47±8.05 vs. 249.48±6.86, P < 0.01 ), and it lied in cytoplasm. In GW9662 pretreatment group the expression of NF-κBp65 ( gray value ) showed no significant difference as compared with that of LPS challenged group ( 214.84±7.91 vs. 249.48±6.86, P>0.05 ).②Coagulation:compared with control group, PT and APTT were significantly prolonged, FIB was significantly decreased, and D-dimer was significantly increased in LPS challenged group [ PT ( s ):18.32±2.03 vs. 12.22±1.38, APTT ( s ):40.05±2.72 vs. 26.64±2.73, FIB ( g/L ): 1.65±0.51 vs. 3.60±0.37, D-dimer ( mg/L ): 2.58±0.73 vs. 0.37±0.06, all P < 0.01 ]. Compared with LPS challenged group, APTT and PT were significantly shortened, FIB was significantly increased, and D-dimer was significantly lowered in ROSI pretreatment group [ PT ( s ):13.93±1.67 vs. 18.32±2.03, APTT ( s ):30.29±0.86 vs. 40.05±2.72, FIB ( g/L ):3.18±0.69 vs 1.65±0.51, D-dimer ( mg/L ):0.40±0.12 vs. 2.58±0.73, all P<0.01 ]. All parameters in GW9662 pretreatment group showed no significant difference as compared with those of LPS challenged group. Conclusions PPAR-γagonist ROSI may ameliorate coagulation disorders in septic rats. PPAR-γ/NF-κB transduction pathway plays an important role in septic coagulopathy.