This thesis aimed at the Bacillus natto TK-1 screened out from Natto.The lipopeptide was purified using Thin-Layer Chromatography(TLC),and investigated its anti-tumor activity.After acid precipitation and methanol extraction,the lipopeptide was separated on TLC.Then the authors get the monomer surfactin which molecular weight is 1036Da through the High performance liquid chromatography(HPLC),electro spray ionization-mass spectrometry(ESI-MS) and infrared(IR).MTT method was implied to testify the anti-tumor activity of the purified sample from TLC.The results indicated a concentration and time-dependent relationships.After 48h,their IC50 were 40 mg/L.The detection with inverted microscope fluorescence microscope displays that the surfactin will cause a series of Morphological changes to the cells.In TUNEL experiment,the authors noticed that surfactin has the ability to induce apoptosis,besides this inhibition shows an obvious time-dependent relationship.

The study was to develop cis-dichlorodiammineplatinum(DDP)-loaded formulations using a novel type of self-assembled compound composed of block copolymers synthesized by poly(?-glutamic acid)(?-PGA).For the potential of targeting liver cancer cells,D-galactose was conjugated on the prepared ?-PGA.In vitro,DDP can be released from the resulting conjugate in PBS:there was a burst release during the first 8 h,then followed by sustained release.DDP could be easily incorporated into poly(?-glutamic acid)-D-galactose esterifiable derivative through a covalent bond.The yield of DDP incorporation into the esterifiable derivative was 9.4%～10.2%.In vitro experiments conclusively established that the poly(?-glutamic acid)-D-galactose esterifiable derivative-Cisplatin Complex Compound(?-D+-DDP)was much less toxic to normal cell lines than DDP only.The surviving rate of cells treated with ?-D+-DDP compound is higher than those treated with free DDP.Also it has obvious antitumor efficiency on human liver tumor BEL-7402 cells.HE staining indicated that the ?-D+-DDP compound make the BEL-7402 apoptosis.These results indicated that the conjugation of DDP to the esterifiable derivative reduced its cytotoxicity activity,but retains its antitumor activity in vitro.In conclusion,the ?-D+-DDP compound could be used as a potential clinic antitumor drug.The ?-PGA obtained by fermentation can be used as a valuable drug carrier system.

DDP could be easily incorporated into poly (?-glutamic acid)-D-galactose esterifiable derivative through a covalent bond. The yield of DDP incorporation into the ?-PGA was 9.4%～10.2%. The DDP was released in the initial 8h in a burst manner,and thereafter in a sustained manner. The results that the conjugation of DDP to poly (?-glutamic acid)-D-galactose esterifiable derivative not only reduced the toxicity of the DDP but also enhanced antitumor activity and the targeting ability. The vivo experiments conclusively established that the ?-D+-DDP compound was much less toxic to animals than DDP alone. A direct evaluation showed that mice treated with ?-D+-DDP compound at a dose of 7.5 mg/kg displayed significant tumor regression. Furthermore,the implanted solid tumors disappeared completely from 35% of the H22 tumor-bearing mice after ?-D+-DDP compound administration. The aforementioned results of biodistributions of the prepared ?-D+-DDP compound in various organs in normal mice demonstrated that the ?-D+-DDP compound had a specific interaction with liver's parenchymal cells and H22 hepatocellular carcinoma tumor cells via ligand receptor recognition. In conclusion,the results indicated that the ?-D+-DDP compound prepared can effectively target the site of hepatoma tumor via the recognition and significantly reduce its size. The ?-D+-DDP compound was less toxic than the free DDP,and could effectively reduce xenografted H22 hepatocellular carcinoma cells in KM mice and prolong the survival of KM mice grafted with H22 hepatocellular carcinoma tumor cells. Therefore,the prepared ?-D+-DDP compound may be used as a potential drug delivery system for the targeted delivery to liver cancers or other liver diseases.

Objective To investigate expressions of phosphorylated c-Jun N-terminal kinase (p-JNK)and P38 mitogen-activated protein kinase(p-P38MAPK)in psoriasis vulgaris lesions. Methods Tissue specimens were obtained from lesions of 30 patients with psoriasis vulgaris and normal skin of 30 healthy human controls. An immunohistochemical study and Western-blot analysis were performed to measure protein expressions of p-JNK and p-P38MAPK in these skin specimens. Results As the immunohistochemical study showed, the expressions of p-JNK and p-P38MAPK(expressed as the average optical density [AOD]value for targeted proteins)were significantly higher in psoriasis vulgaris lesions than in normal skin tissues (p-JNK: 0.663 ± 0.016 vs. 0.333 ± 0.009, t = 44.869, P < 0.001; p-P38MAPK: 0.436 ± 0.011 vs. 0.306 ± 0.010, t = 21.913, P < 0.001). Western-blot analysis also showed increased protein expressions of p-JNK and p-P38MAPK in psoriasis vulgaris lesions compared with normal skin tissues (t = 20.477, 165.084, respectively, both P <0.05). Conclusion The activation of JNK and P38MAPK may be involved in the overproliferation of epidermal cells in psoriasis vulgaris lesions.

Adult ; Factor VII Deficiency ; complications ; Female ; Humans ; Liver Cirrhosis, Biliary ; complications

OBJECTIVETo evaluate the results of the surgical treatment of patients with Budd-Chiari syndrome (BCS).

METHODSThe clinic data of 120 BCS patients who underwent various consecutive surgical treatments from July 2001 to October 2010 was analyzed. There were 82 male and 38 female patients, aging from 11 to 72 years with a mean age of (41 ± 13) years. All patients experienced various examinations to identify the pathological type of BCS. There were 5 cases of small hepatic veins type, 28 cases of large hepatic veins (LHV) type, 31 cases of inferior vena cava (IVC) type and 56 cases of combined obstruction of LHV and IVC. Totally, 25 patients experienced interventional treatment, include percutaneous transluminal angioplasty and/or stenting for stenosis of hepatic vein and/or IVC, 77 patients experienced open-thorax operation for BCS radical resection under protection of right atrium by-pass with extracorporeal circulation.

RESULTSTotally 97 cases were followed up from 1 to 120 months after various surgical treatment methods. Perioperative mortality was 6.2% (6/97). Follow-up period mortality was 8.2% (8/97). The restenosis of IVC and/or hepatic vein happened in 3 cases out of 25 cases in intervention treatment group in contrast with 15 cases out of 77 cases in radical resection group. The 5-year patency and survival rate of IVC/hepatic vein were 64.5% and 83.3%.

CONCLUSIONSThe surgical treatment of BCS need to get accurate diagnosis and pathological classification firstly, then, to choose appropriate therapeutic strategies based on individual pathological classification. The BCS radical resection can be an alternative method in some particular pathological classifications and the cases who failed in interventional treatment.

Adolescent ; Adult ; Aged ; Budd-Chiari Syndrome ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo summarize clinical experience in the treatment of low rectal cancer with anterior perineal plane for ultra-low anterior resection of the rectum(APPEAR).

METHODSClinical and follow-up data of 26 patients with low rectal cancer undergoing the APPEAR operation in Department of General Surgery at Peking Union Medical College from September 2009 to August 2011 were retrospectively analyzed.

RESULTSThe 26 cases consisted of 19 male and 7 female patients with an average age of (63.4 ± 9.5) years. The average tumor distance from the anal verge was (4.6 ± 0.7) cm according to preoperative examinations. Fourteen patients received neoadjuvant radiochemotherapy before the operation. All the 26 patients had successful sphincter-preserving operations. The average operative time was (170 ± 21) min and the average intra-operative blood loss (140 ± 69) ml. Complications included one case of intraoperative injury to the rectal wall and 4 cases of postoperative perineal wound infection. Postoperative pathological examination showed well to moderately differentiated adenocarcinomas(n=10), moderately differentiated adenocarcinomas with partial mucinous adenocarcinomas (n=7), poorly differentiated adenocarcinoma(n=1), villous adenoma with high-grade intraepithelial neoplasia (n=1), and rectal villous adenoma(n=1). In 6 cases no residual tumor cells were detected in the surgical specimens. All the patients were followed-up for an average period of(11.4 ± 5.6) months. No impaired urinary function or tumor recurrence was observed during the follow-up. Eighteen patients had the transverse colon stoma closure six months after the operation. The average Wexner continence score was 5.5 after colostomy reversal surgery. The anorectal manometry tests showed that maximum squeeze pressure of the anal sphincter was(224.0 ± 59.3) mm Hg. The maximum resting pressure was (42.5 ± 11.8) mm Hg, and the maximum tolerable volume of the rectum was (120.0 ± 27.4) ml. Anorectal reflexes were present in all these patients.

CONCLUSIONThe APPEAR technique can be applied in the sphincter-preserving operations for low rectal cancer patients with satisfactory anal function.

Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Perineum ; surgery ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Retrospective Studies ; Treatment Outcome

AIM To establish a UFLC-QTRAP-MS/MS method for the simultaneous content determination of phenylalanine,isoleucine,methionine,valine,proline,tyrosine,aspartic acid,histidine,arginine,lysine and glutamic acid in Gynostemma pentaphyllum (Thunb.) Makino.METHODS The analysis of aqueous extract of G.pentaphyllum was performed on a Waters XBridge Amide column (2.1 mm × 100 mm,3.5 μm),with the mobile phase comprising of water-acetonitrile (containing 0.2％ formic acid) flowing at 0.6 mL/min in a gradient elution manner.RESULTS Eleven amino acids showed good linear relationships within their own ranges (r ＞0.998 9),whose average recoveries were 90.74％-103.05％ with the RSDs of 0.61％-5.00％.CONCLU-SION This accurate,stable and reproducible method can be used for the quality control of G.pentaphyllum.

Gastrointestinal microbiota has an important impact on physiological functions of human body. In recent years,the importance of gastrointestinal microbiota in tumorigenesis has emerged. Aberrant location and proportion of microbiota can cause not only carcinogenesis of local epithelia,but also tumorigenesis in submucous tissues or even distant organs. Certain microbes can produce genotoxin or generate reactive oxygen species to cause DNA damage and affect DNA damage repair,which leads to genetic instability and induces cell transformation. Moreover,gastrointestinal microbiota is able to affect the function of host immune system and form an immunosuppressive microenvironment,while some other pathogenic bacteria can cause chronic inflammation and may be involved in tumor immune escape. Based on the above,gastrointestinal microbiota is oncogenic and closely linked to tumorigenesis.

Tongue diagnosis is one of unique techniques of TCM diagnosis. Its subjective and empirical description of written words, spoken language and pictures have been used until now, however, which cannot express the complete information of tongue image comprehensively and objectively. The development of modern technology provides new ideas and methods for the study of tongue diagnosis in TCM. For example, the application of spectroscopy is of great significance for restoring tongue image information. This article briefly introduced the spectroscopy method, summarized its research and application in the field of TCM tongue diagnosis, and put forward the problems and prospects to provide references for further research.