The occurrence, development and outcome of most diseases are a complex process. It is the advantage of traditional Chinese medicine to establish complex interventions that are consistent with the characteristics of disease development. The specific steps are as follows: quantitative research on literature research, establishment of interview framework; focus on interviews, specificization of interview outlines; semi-structured interviews, preliminary construction of complex intervention programs; evaluation of efficacy of complex interventions. The introduction of semi-structured interviews and other qualitative research methods into the modernization of traditional Chinese medicine, combined with quantitative methods such as text analysis and data mining, is also helpful in formulating the standard of diagnosis and treatment system with Chinese characteristics.

OBJECTIVEThe study was designed to investigate the expression patterns of metalloproteinase (MMP)-13 protein in invasive breast carcinoma and to determine the clinicopathological and prognostic values of its various localization and relation to the tumor phenotypes.

METHODSImmunohistochemistry was performed on paraffin-embedded tissue array from 263 invasive breast carcinomas to investigate the protein expressions of MMP-13, estrogen receptor, progesterone receptor, HER2, MMP-2, MMP-9 and tissue inhibitor of matrix metalloproteinases (TIMP)-1, TIMP-2.

RESULTSMMP-13 protein was detected in the cytoplasm of carcinoma cells and peritumoral fibroblasts. High level expression of MMP-13 protein in tumor cells was associated with more lymph node involvement and higher tumor grade (both P < 0.01), and positively correlated with HER2 (P = 0.015) and TIMP-1 protein (P < 0.01) expression in carcinoma cells. Moreover, high expression of MMP-13 was associated with shortened overall survival for the entire patient population and the patient group with positive lymph node. Tumor cell derived MMP-13 had different impact on patients with different HER2 status. Peritumoral fibroblasts derived MMP-13 protein, although correlated with tumor cell derived MMP-13 and associated with lymph node stage and HER2 expression, was found having less prognostic impact. Univariate survival analysis showed that the tumor size, grade, lymph node status, PR status, HER2 expression, tumors TIMP-1 and MMP-13 expression were prognostic factors. However, multivariate survival analysis showed that only tumor size, lymph node status, HER2 expression, tumors TIMP-1 and MMP-13 were independent prognostic factors.

CONCLUSIONMMP-13 protein expressed by tumor cells correlates with the invasion and metastasis of breast carcinoma, and therefore, may serve as a poor prognostic marker for the patient.

Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; physiopathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Lymph Nodes ; pathology ; Matrix Metalloproteinase 13 ; analysis ; genetics ; Matrix Metalloproteinase 9 ; analysis ; Neoplasm Invasiveness ; diagnosis ; physiopathology ; Neoplasm Staging ; classification ; Prognosis ; Receptor, ErbB-2 ; analysis ; Receptors, Estrogen ; Receptors, Progesterone ; analysis

OBJECTIVESTo investigate the effects of newborn bull serum(NBS), vitamin C and vitamin E on cryopreservation of mouse seminiferous epithelial cells.

METHODSThe seminiferous epithelial cells from 7-day-old mice were cryopreserved in different freezing solutions. The cell recoveries were examined by Trypan blue exclusive staining after thawing. The freezing solutions composed of DMEM, 10% dimethylsulphoxide(DMSO), and 0, 5%, 10%, or 20% NBS, respectively, or composed of DMEM, 10% DMSO, 10% NBS, and 150 micrograms/ml vitamin C or 50 micrograms/ml vitamin E, respectively.

RESULTSThe cell recoveries in freezing solution containing 0, 5%, 10%, or 20% NBS were 83.4%, 84.7%, 85.7% and 83.6%, respectively. There were no significant differences between them. The cell recoveries in freezing solution containing vitamin C or vitamin E were 88.0% and 82.9%, respectively. There was no significant differences compared with that in freezing solution containing 10% DMSO and 10% NBS.

CONCLUSIONSNBS, vitamin C and vitamin E have no significant protecting effects on mouse seminiferous epithelial cells, and can not significantly improve the cell recoveries.

Animals ; Ascorbic Acid ; pharmacology ; Cattle ; Cryopreservation ; Epithelial Cells ; physiology ; Fetal Blood ; physiology ; Male ; Mice ; Seminiferous Epithelium ; cytology ; Vitamin K ; pharmacology

OBJECTIVETo probe into indication of living-related liver transplantation (LRLT) for Wilson's Disease.

METHODSFrom January 2001 to February 2007, thirty-seven living-related liver transplants were performed. A retrospective analysis was carried on outcome of those patients. The indications for LRLT were acute hepatic failure in 3 patients and chronic advanced liver disease in 32 patients including 13 patients with Wilsonian neurological manifestations. Two patients presented with severe Wilsonian neurological manifestations even though their liver functions were stable. According to the scoring system for evaluation of the neurological impairment in Wilson disease based on neurological signs and functions (total score was 30), the pre-transplantation score of those patients with neurological manifestations was 15.9 +/- 4.3 (n = 15).

RESULTSThirty-seven patients were followed up for 20 - 93 months. The survival rates of post-transplant patients and grafts at 1, 3, and 5 year were 91.9%, 83.8%, 75.7%, and 86.5%, 78.4%, 75.7%, respectively. Postoperative surgical complications occurred in 2 donors with bile leakage required drainage, in 2 recipients with hepatic thrombosis underwent retransplantation of cadaveric liver and in 1 recipient with hepatic stenosis required balloon dilatation. Neurological function was improved in all recipients and the score of posttransplantation at 6, 12, 18, 24, and 30 month was 17.5 +/- 3.7 (n = 13); 21.0 +/- 4.3 (n = 12); 23.9 +/- 3.9 (n = 10); 26.6 +/- 2.2 (n = 10) and 28.1 +/- 1.9 (n = 7) respectively.

CONCLUSIONSPatients with acute hepatic failure or patients with severe liver disease unresponsive to chelation treatment should be treated with LRLT. Early transplantation in patients with an unsatisfactory response medical treatment may prevent irreversible neurological deterioration even though their liver function is stable.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Hepatolenticular Degeneration ; complications ; surgery ; Humans ; Liver Failure ; etiology ; surgery ; Liver Transplantation ; Living Donors ; Male ; Nervous System Diseases ; etiology ; Retrospective Studies ; Transplantation, Homologous ; Treatment Outcome ; Young Adult

OBJECTIVEViral hepatitis remains a major public health problem and the most common type of liver disease worldwide. There are an increasing number of patients with chronic hepatitis B who develop acute hepatitis on chronic condition (AOC) and die of acute hepatic failure both as a result of lack of understanding of the pathogenesis of the disease and lack of effective treatment. The hallmark of AOC is the extreme rapidity of the necromicroinflammatory process resulting in widespread or total hepatocellular necrosis in weeks or even days. Our previous studies have shown in an experimental animal model of fulminant viral hepatitis caused by murine hepatitis virus strain 3, the importance of macrophage activation, and expression of a unique gene mfgl 2 which encodes a serine protease capable of directly cleaving prothrombin to thrombin, resulting in widespread fibrin deposition within the liver and hepatocyte necrosis. The undergoing study in this report is designed to identify the role of hfgl 2 (human fibrinogen like protein 2) /fibroleukin in patients with viral hepatitis.

METHODSLiver tissues were obtained from 23 patients with AOC hepatitis B, and from 13 patients with inactive chronic hepatitis B (CHB) and 14 patients with chronic hepatitis B with cirrhosis during the year of 1995 to the end of 2001. Liver biopsies were performed within 30 min after the patients were diagnosed with death as a result of acute hepatic failure. Liver samples were also obtained from 4 liver donors as normal controls. In addition, peripheral blood mononuclear cells (PBMC) were isolated from 30 patients (unpaired) with AOC hepatitis B and 10 patients with CHB during the May of 2001 to March of 2002 and 10 healthy volunteer as negative control. PBMCs were freshly isolated and smeared on slides and kept at -80 degree C for further use. Histological sections were stained with hemotoxylin and eosin. A 169 bp of hfgl 2 cDNA probe and a polyclonal or monoclonal antibody against hfgl 2 were used to detect the expression of hfgl 2 mRNA and protein in liver samples as well as PBMC by immune histochemistry separately.

RESULTSLiver tissues from the patients with acute on chronic hepatitis had classical pathological features of acute necroinflammation. Hfgl 2 was detected by immune histochemistry in 21 of 23 patients (91.3%) in liver sections from patients with acute on CHB, while only 1 of 13 patients (7.7%) with CHB and cirrhosis and no evidence of active disease had hfgl 2 mRNA or protein expression. 28 of 30 patients (93.3%) with acute on CHB and 1 of 10 with CHB were detected with hfgl 2 expression in PBMC. There was no hfgl 2 expression in either the liver tissue or the PBMC from the normal donors. There was positive correlation of hfgl 2 expression and the severity of the disease displayed by the value of bilirubin and PT.

CONCLUSIONThe molecular and cellular results reported here in patients with acute on chronic hepatitis and who died of acute hepatic failure correlates with previous report in 8 patients with fulminant hepatic failure (FHF) and mimic closely the changes observed in the murine model of fulminant viral hepatitis in which the pathogenesis of the disease has been studied in a stepwise fashion. This study further suggests that virus induced hfgl 2 prothrombinase/fibroleukin expression and the potent function of the protein it encodes plays a pivotal role in initiating acute severe hepatitis on the baseline of chronic hepatitis. The measurement of hfgl 2/fibroleukin expression in PBMC may serve as a useful marker to monitor the severity of disease in patients with the AOC hepatitis B and a target for therapeutic intervention.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Fibrinogen ; biosynthesis ; genetics ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; Liver ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; RNA, Messenger ; biosynthesis ; genetics ; Severity of Illness Index ; T-Lymphocytes ; metabolism

OBJECTIVETo evaluate the outcome of emergency adult right lobe living donor liver transplantation for fulminant hepatitis.

METHODSNine cases of adult right lobe living donor liver transplantation were performed from September 2002 to August 2005, the clinical and follow-up data was analyzed.

RESULTSAccording to Child Pugh Turcotte (CPT) classification, 9 patients were classified as grade C before transplant. The Model for End-Stage Liver Disease (MELD) scores of these patients were 26.7 +/- 8.8. The principal pre-transplant complications included hepatic encephalopathy (5 cases), electrolyte disturbance (3 cases), renal failure (2 cases), gastrointestinal bleeding (1 case). The operations in donors and recipients were all successful. The post-transplant complications induced pulmonary infection in 2 patients, acute renal failure in 3 and transplantation related encephalopathy in 1. There were no primary graft non-function and no blood vessel and bile tract complications occurred. One-year survival rate was 55.6%. No serious complication or death found in donors.

CONCLUSIONSEmergency adult to adult living donor liver transplantation is an effective treatment for fulminant hepatitis but the safety of the donors should be assessed strictly preoperation.

Adult ; Critical Illness ; Emergency Medical Services ; Female ; Follow-Up Studies ; Hepatitis ; pathology ; surgery ; Humans ; Liver Transplantation ; methods ; Living Donors ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate and evaluate different surgical methods applied in living-donor liver transplantation (LDLT).

METHODSFifty patients with end-stage liver disease received LDLT in our department between January 1995 and March 2006. The data were analyzed on a retrospective basis. The choice of different surgical methods, strategies applied to ensure the safety of donors and indications of LDLT in the series were reviewed.

RESULTSAll donors recovered uneventfully. Among the 50 patients, 47 recipients presented with end-stage cirrhosis, 3 patients suffered from malignant tumor. To date, 6 recipients died after LDLT, among them, 3 recipients died of the operation and the other 3 recipients died of long-term complications. Resected donor livers included 9 cases of segments V, VI, VII and VIII (not including the middle hepatic veins) and 1 case of segments V, VI, VII and VIII (including the middle hepatic veins), 36 cases of segments II, III and IV (including the middle hepatic veins) and 4 cases of segments II, III, and part of IV (not including middle hepatic veins).

CONCLUSIONSLDLT helps tackle the problem of donor shortage in the world. The process is complicated, and it is very important to choose appropriate surgical methods for the improvement of surgical achievement and donor safety.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Liver Cirrhosis ; surgery ; Liver Neoplasms ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo immunopurify human endometrial endothelial cells (HEEC) from fresh surgical specimens of endometrial cancers and normal endometrial tissues, and investigate their biological characteristics.

METHODSEndothelial cells of endometrial cancers and normal endometrial tissues were isolated using anti-CD31 conjugated magnetic microbeads. The isolated endothelial cells were cultured in vitro and their origins were identified. Their angiogenic characteristics were observed by MTT, wound healing, Transwell cell invasion and tube formation assays.

RESULTSFlow cytometry revealed that the immunopurification technique yielded endothelial cell purity of > 95% in all samples. All purified HEEC were characterized as endothelial cells on the basis of expression of the classical endothelial markers vWF and CD31 as shown by immunofluorescence examination. Although the tumor-associated HEEC didn't show more rapid proliferation than normal HEEC, they exhibited enhanced migration ability (P = 0.006), potent invasiveness (P = 0.033), and elevated tube formation in vitro (P = 0.029).

CONCLUSIONSHuman endometrial endothelial cells can be efficiently isolated from endometrial cancer and normal endometrial tissues by immunomagnetic methods. Tumor-associated HEEC exhibit enhanced migratory ability, potent invasiveness, and elevated tube formation in vitro.

Adult ; Aged ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Endometrial Neoplasms ; metabolism ; pathology ; Endometrium ; cytology ; metabolism ; pathology ; Endothelial Cells ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo express and purify HBoV VP2 protein, and the monoclonal antibody against HBoV VP2 protein was prepared with hybridoma technique.

METHODSThe HBoV VP2 cloned into vector pET-30a was expressed in E. coil. After purified by immobilized metal affinity chromatography, the BALB/c mouse was immunized with purified protein as antigen. The positive hybridoma cells were screened with hybridoma technique and ELISA assay. Isotype and titer of the monoclonal antibody were detected.

RESULTSThe recombinant HBoV VP2 protein was expressed and purified, and then the monoclonal antibody was obtained with hybridoma technique. The titer of the IgG monoclonal antibody was up to 1:4 x 10(5).

CONCLUSIONMonoclonal antibody against recombinant HBoV VP2 protein was prepared and the antibody titer was high. This work may provide a new method in rapid diagnosis and study of HBoV.

Animals ; Antibodies, Monoclonal ; immunology ; Capsid Proteins ; genetics ; immunology ; Human bocavirus ; immunology ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Proteins ; biosynthesis ; immunology ; isolation & purification

OBJECTIVETo study the significance of supratubal recess and its aeration pathway to epitympanum in the pathogenesis of cholesteatoma otitis media.

METHODSFifty-two ears of cholesteatoma were selected as study group. Sixteen ears of traumatic facial palsy with pneumatic mastoid, which had no history of chronic otitis media were selected as control group. The status of supratubal recesses of all and their aeration pathways to epitympanum were observed in operations.

RESULTSSixteen ears from control group clearly presented supratubal recesses. Membrane closure was founded in four of them. The aeration pathways of fifty-two ears (100%) from study group were all completely closed. Comparing with control group, the difference was obviously significant (chi2 = 41.7144, P = 0.000). Among these cases, bony closure was observed in thirty-four ears (65.4%), while membrane closure in eighteen ears (34.6%). Their epitympanum space was very narrow and mastoid was sclerotic or poorly developed.

CONCLUSIONSBlockage of the aeration pathway between supratubal recess and epitympanum was possible one of the origins of negative-pressure status of epitympanum and mastoid, which might lead to the formation of aural cholesteatoma. Anatomy variation of the aeration pathway from supratubal recess to epitympanum might be a pathogenesis factor of cholesteatoma otitis media. It suggested that opening the aeration pathway in tympanoplasty with intact canal wall up technique might be helpful to prevent recurrence of aural cholesteatoma.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Cholesteatoma, Middle Ear ; pathology ; Eustachian Tube ; pathology ; Facial Paralysis ; pathology ; Female ; Humans ; Male ; Middle Aged ; Young Adult