Objective To investigate the feasibility of sclerosing agent injection in the treatment of benign thyroid cysts. Methods Clinical data of 60 patients with thyroid cysts, who had been treated by injection of absolute alcohol or 10% sodium chloride solution, were reviewed retrospectively. Results A total of 74 lesions existed in 60 patients, and 182 times of injection were performed, with a total cure rate of 98 3% (59/60). No complications occurred. Follow-up observations for 1～5 years (mean 2 years) found no recurrence. Conclusions Sclerosing agent injection is a feasible method for benign thyroid cysts, if an exclusion of cystic papillary carcinoma is made.

Objective: To investigate the effects of combined use of early enteral nutrition, growth hormone and fibrin glue on early healing of intestinal anastomosis in intra-abdominal sepsis rats. Methods: Male Wistar rats were randomized into 3 groups. In Control group, antimesenteric transverse enterotomy of 4mm was performed, a second laparotomy was subsequently performed after 24 hours, 6cm of the intestine in the middle of the fistula segment was resected. An end-to-end single layer anastomosis was constructed using 8 interrupted 6-0 polypropylene sutures; In EN group, anastomosis was constructed using 4 interrupted sutures and sprayed fibrin glue, enteral nutrition was given on the second postoperative day for 12 days; In EN/GH group, the animals were treated as EN group, and growth hormone(3.0 U/kg/d) was injected subcutaneously every 24 hours for 14 days. The anastomotic bursting pressure and breaking strength, hydroxyproline content and histology were measured as indicators of wound healing. Results: Anastomotic bursting pressure, anastomotic breaking strength and hydroxyproline content in EN+GH group were higher than that in control and EN groups. Conclusions: Combined use of early enteral nutrition , growth hormone and fibrin glue improves early healing of intestinal anastomosis in intra-abdominal sepsis rats.

Objective To propose the developmental orientation of the science of pathology in PLA in the next five years by reviewing the advances and developmental tendency of pathology in the Eleventh Five-Year Plan. Methods The latest progresses and developmental tendency in pathology were reviewed by reviewing the related reviews and treatises published domestically and abroad. Results With the scientific and technological progresses,especially rapid development of molecular biology,a lot of new knowledge,theories,techniques and methods had been proposed,established and applied in various fields of pathology successfully,which provided a new opportunity for improving clinical pathological diagnosis,pathologic research and teaching,as well as cultivation of academic talents of pathology discipline. Meanwhile,these new advances had also broadened the new field of pathological studies and accelerated the development of military pathology. Remarkable achievements have been obtained in military pathology and oncological pathology,etc. Conclusion Emphasis should hereafter be put on the researches in the fields of molecular pathology,military pathology,clinical pathology and army-civilian common pathological techniques,so as to raise the technical level of pathological diagnosis and perfect the construction of hospital pathology discipline.

Object To establish a method for the assay of myricetin in Ampelopsis grossedentata (Hand. Mazz.) W. T. Wang. Methods The determination was carried out with RP HPLC on Nova pak C 18 stainless steel column (150 mm ? 3 9 mm) with methanol∶water (40∶60) as the mobile phase, and detected at a UV wave length of 254 nm. Results The coefficient of variation was 2.763% with average recoveries=97.4%. The minimal detectable limit was 15 ng. Contents of myricetin in different parts of A. grossedentata from Hunan Province varied from 1.57% to 2.17%. Conclusion The method is rapid, simple, accurate and good for the determination of myricetin in A. grossedentata.

ObjectiveTo investigate the effects of splenectomy on the intestine mucosa barrier in rats with obstructive jaundice. Methods50 Wistar rats were divided randomly into the obstructive jaundice group (OJ), in which the animals underwent operation to ligate common bile duct, and the obstructive jaundice + splenectomy group (OJ+ S). Seven days post-operation, plasma endotoxin levels were detected. Intestinal mucosa permeability was measured by the ratios of lactulose and mannitol (L/M). Immunohistochemistry and Western blot were used to examine the expression of tight junction proteins (ZO-1 and occludin) in the distal ileum mucosa. Western blots images were analyzed quantitatively. ResultsAverage ratios of L/M and plasma endotoxin were decreased obviously in the OJ+S group compared to those in the OJ group (all P=0. 001). Compared with the OJ group, the average intestinal villus height and mucosa thickness were upgraded somewhat in the OJ + S group (P = 0.019, 0. 001 ). By immunohistochemistry staining seven days post-operation, same comment as above the amounts of strong positive expression of ZO-1 were significantly decreased in the OJ group (6/18, P-0. 021). There wewas no difference between the OJ+S group(8/17) and the OJ group.The amount of strong positive expression of occludin was higher in the OJ + S group than that of the OJ group(10/17 vs 4/18, P= 0. 026). The same outcomes were obtained by quantitative Western blot images. Conclusion The intestinal epithelial permeability was increased in rats with obstructive jaundice,and intestinal barrier was damaged. After excising spleen, the amount and distribution of tight junction proteins were changed and the impairment of intestinal barrier was abated.

Objective To introduce the mutated gene coding cysteine into the gene of dsFv VL of human antibody to N terminal fragment of lipopolysaccharide binding protein(LBP)and to express,purify the mutated dsFv VL in bacterium.Methods We reconstructed and sequenced the mutated gene of VL of human mAb Fab to LBP by Mega-primer PCR based on point mutagenesis method.Some codes of FWR1 of VL had been replaced by TGT in order to code cysteine.The DNA sequence of reconstructed VL was inserted into vector pET-28a(+),then VL was expressed by E.coli.BL21 star(DE3)and was purified by chromatography.Finally the activity of VL to bind NH-LBP was determined by ELISA.Results The results showed that the cysteine was introduced into the position 21 amino acid of VL to replace the threonine.The gene of VL was about 650 bp and relative molecular weight of VL was 28?103.VL could bind NH-LBP directly.Conclusion These have laid a foundation for producing the dsFv against NH-LBP.

Objective To investigate the effects and mechanisms of bile on small intestine mucosal barrier.Methods Fifty Wistar rats were assinged into 3 groups randomly: obstructive jaundice(OJ) group(n=20),biliary external drainage group(n=20) and control group(n=10).Ten days after operation,the plasma endotoxin level was determinated,the terminal ileum mucosas was obtained to be morphologically measured by light microscope,and immunohistochemistry and Western blot were uesd to examine the expressions of tight junction proteins zona occludens-1(ZO-1) and occludin in the mucosas.Results Atrophy significantly appeared in the distal ileum mucosas in OJ group.Compared with control group,the intestinal villus height,mucosa thickness and crypt depth in OJ group were obviously decreased 27.8%,21.7%,and 25.4%(P=0.001,0.001,0.040).There were no differences between external drainage group and control group(P=0.050,0.070,0.080);While the values of external drainage group were significantly higher than those in OJ group(all P=0.001).The level of plasma endotoxin was up to(1.49?0.27) EU/ml in OJ group compared with control group((0.27?0.09) EU/ml),P=0.001.In external drainage group,the value was(0.91?0.25) EU/ml,which was obviously higher than that in control group and lower than that in OJ group(all P=0.001).Immunohistochemical study showed strong positive expression of ZO-1 dropped from 7/10 in the control group to 6/20 in OJ group(P=0.040),occludin expression was 8/10 in control group and 7/20 in OJ Group(P=0.020);expressions of them in external drainage group(8/20(P=0.100,0.210) and 9/20(P=0.060,0.200)) displayed no significant differences compared with the other twogroups.Quantitative testing of Western blot showed the expressions of ZO-1 and occludin in OJ group were significantly lower than those in control group(P=0.001,0.010),the values in external drainage group were higher than those in OJ group(P=0.005,0.014).The expression of ZO-1 was lower in external drainage group than that in control group(P=0.001),and there was no significant difference of occludin between the two groups(P=0.062).Conclusion Lack of intestinal bile will undermine the intestinal tight junction protein composition,and make intestinal mucosal barrier impaired.The intestinal barrier more severely injured when biliary tract obstructs because of multiple factors.Bile plays an important role in the maintenance of intestinal mucosal barrier.

Objective:To generate monoclonal antibodies against ciprofloxacin (CIP) and to analyze its immunological traits for establishing a determination method for ciprofloxacin residues in food tissues.Methods:BALB/c mice were immunized by the conjugation of CIP-BSA successfully.The splenic cells of BALB/c mice and the oncocyte were fused and screened in HAT culture medium.Cell strains of 1C9,3F6,6H2,6A7,6G11 and 8F5 were cloned,which could secret the monoclonal antibody (Mab) for ciprofloxacin.Results:By ELISA method,the haplotype of Mabs were identified.The antibody secreted by 1C9,3F6 and 6A7 were classified to the IgG2a.Those of 6H2 and 8F5 were classified to the IgG1 while 6G11 to IgG3.The results of SDS-PAGE showed that the Mab protein was made up of the weight chain and the light chain whose molecular weight were 50 kD and 25 kD,respectively.All of those Mabs had fine specificity and sensibility.The indirect ELISA titer of 1C9 was 1:6.4?102 in supernatant and 1:5.6?105 in ascites.The affinity of 1C9 was 2.85?109 L/mol and the value of IC50 reached to 245.86 ng/ml.The protein secreted by 1C9 was screened for establishing the ELISA method and the detective limitation reached to 45.25 ng/ml.There were no cross reactions to Ofloxacin,Difloxacin,Sarafloxacin,Malachite green,Chloramphenicol and Furacilin while the rate of cross reaction to Enrofloxacin could reach to 84.6%.Conclusion:The Mabs are sensitive and specific,which are suitable to be applied in establishing immunoassay of CIP residues.

Objective: To investigate the mechanism of intestinal anastomosis in a rat model of intra-abdominal sepsis.Methods:Male Wistar rats were randomized into 2 groups.Control rats underwent intestinal anastomosis.Sepsis group underwent intestinal anastomosis after intestinal fistula operation.fibronectin(FN),?-smooth muscle actin(?-SMA),vascular endorthelial growth factor(VEGF) and apoqtosis in anastomoses were measured.Results:FN,?-SMA,VEGF in control group were higher than sepsis group and apoptosis were lower than the other.Conclusion:It is indicated that the repair of tissue was out of balance after infection,and the organization repair was retarded.The collagenoblast and the epithelial cell migration were slowed down,and the blood capillary regeneration was slowly.Not only the granulation organization was reduced,but also apoptosis was prematurely appeared.

Objective To reconstruct the proteinic sequence of human cathelicidin LL-37 to increase the bactericidal activity of LL-37 and to express the reconstructed LL-37 (rLL-37) in bacterium. Methods The two dimensional structure, three dimensional structure and chemical characteristic of LL-37 were analyzed by Soft Ware Anthepro 5.0 and SWISS-MODEL. Without the three dimensional changes of LL-37, some negative amino acids of human cathelicidin LL-37 were replaced by positive amino acids and the positive charge of LL-37 was increased. According to the proteinic sequence changes of rLL-37, the DNA sequence of rLL-37 was reconstructed by Touch-Down PCR and recombined with vector pET-28a (+), thus rLL-37 was expressed in E.coli. BL21 (DE3) by the induction of IPTG and was purified by chromatography. Results Glu~ 16 , Asp~ 26 , Glu~ 36 of LL-37 were replaced by Gln~ 16 , Asn~ 26 , Gln~ 36 and the static charge of LL-37 was increased from +5.8 to +9.0 at pH 7.4. The DNA sequence of rLL-37 was reconstructed and inserted into vector pET-28a (+), the rLL-37 was expressed in E.coli. BL21 (DE3) and purified by strong cation exchange supports Macro-Prep High S successfully. The rLL-37 was proved by the means of inhibitory zone to be able to kill Gram-negative bacteria and Gram-positive bacteria. Conclusion It is feasible to reconstruct human cathelicidin LL-37 and express the protein in bacteria by fusion, which make it possible to produce more rLL-37 and study its biological function deeply.