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 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.

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.

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.

Object To stuay the quality of seedling leaves cultured from improved variety of Ginkgo biloba L.. Methods The content of terpene lactones in seedling leaves of G. biloba were determined by RP-HPLC-ELSD.Results The content of total terpene lactone producted in Jun. was more than that in Aug., and it was obviously lower in Nov.. The content of terpene lactone in grafting was more than that in seedling. The content of terpene lactone in male trees was more than that in female trees.Conclusion The content of the active components was related to the season of growth, age of tree, raising modes, environment of growth, geographic conditions and others. The evaluation of the content of the main active components BB and GB showed that the quality of seedling leaves cultured from improved variety of G. biloba in Jinagxi is better than that reported by literature.

Objective To employ 2 approaches to construct and express reconstructed LL-37 (rLL-37) in procaryotic system, and to explore a better preparation method. Methods The first method: the rLL-37 was inserted into vector pET-28a (+), then was induced to express in E.coli. BL21 (DE3) and purified by chromatography; the second method: the rare codons in the rLL-37 gene sequence were substituted by the preferred codons of procaryotic cell, and a fragment of carrier protein molecule (CPM) was added to the N termination of the objective sequence to construct expression plasmid pET-30a(+)-CPM-rLL-37, then the rLL-37 was expressed in E.coli. BL21 Star(DE3) and purified by chromatography. The productive rates of the 2 methods were compared and the antimicrobial effects of obtained rLL-37 was studied. Results The first method: the DNA sequence of rLL-37 was obtained successively by Touch-Down PCR. The expression plasmid pET-30a(+)-CPM-rLL-37 was expressed with fusion protein in E.coli BL21 (DE3). The expression rate accounted for 20% of total bacterio-protein, then the expressed product was purified by using high positive ion exchange column Macro-Prep High S; The second method: a fragment of carrier protein molecule was designed that contained 28 amino-acid residue and its pHi was 2.7, net charge was-6.0 at pH 7.4. After the expression plasmid pET-30a(+)-CPM-rLL-37 was constructed successively, it was expressed in E.coli BL21 Star (DE3). The expressed fusion protein accounted for 35% of total bacterio-protein, then the expressed product was purified by using affinity binding chromatography with TALON resins successfully. The obtained 2 kinds of rLL-37 were able to kill both Gram-negative and-positive bacteria by the means of inhibitory zone. Conclusion It’s feasible to prepare efficiently rLL-37 in procaryotic system, which founds the basis for the further research on bactericidal activity of rLL-37.

Objective: To investigate the mechanism of combined growth hormone and fibrin glue on early healing of intestinal anastomosis in intra-abdominal sepsis rats.Methods: Male Wistar rats of abdominal sepsis were randomized into 3 groups.Control rats were given intestinal anastomosis after intestinal fistula operation.Glue group was given intestinal anastomosis and fibrin glue after intestinal fistula.GH group was given intestinal anastomosis and fibrin glue and growth hormone after intestinal fistula.FN,?-SMA,VEGF,and apoptosis in anastomoses were measured.Results: Expression of intestinal FN,?-SMA,and VEGF in GH group were higher than those in other groups and apoptosis were lower than others.Conclusion: It is indicated that combined growth hormone and fibrin glue ameliorates healing of intestinal anastomosis in intra-abdominal sepsis rats.

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 obtain the disulfide stabilized Fv fragment (dsFv) against N-terminal fragment of human lip polysaccharide binding protein (NH-LBP) and to identify its biological vitality. Methods The disulfide stabilized Fv fragment antibody (dsFv) was obtained after the inclusion bodies of dsFvVH and dsFvVL had been refolded and purified. Then the characteristics of dsFv were determined in vitro by ELISA and by detecting the secretion of tumor necrosis factor alpha (TNF-?) in rats. Results There was 2.1 mg protein of dsFv obtained. dsFv had good combination with NH-LBP and could restrain inflammatory reaction caused by lip polysaccharide (LPS) in vivo. Conclusion It is feasible to get dsFv against NH-LBP by respective expression of VL and VH. The partial inhibition of the biological function of LBP by dsFv is a new way to restrain the over-inflammatory reaction in vivo.

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.