Adenocarcinoma ; diagnosis ; pathology ; Adult ; Aged ; Aged, 80 and over ; Ascitic Fluid ; chemistry ; Cadherins ; analysis ; Calbindin 2 ; Carcinoembryonic Antigen ; analysis ; Diagnosis, Differential ; Epithelial Cells ; chemistry ; Female ; Humans ; Immunohistochemistry ; Keratin-5 ; analysis ; Male ; Middle Aged ; Pericardial Effusion ; chemistry ; diagnosis ; Pleural Effusion ; chemistry ; diagnosis ; Pleural Effusion, Malignant ; chemistry ; diagnosis ; S100 Calcium Binding Protein G ; analysis

BACKGROUND: Icariin has a broad prospect for promoting cell proliferation. Differentiation direction of periosteal cells is uncertain, but the cells are easy to be induced by ultrasound, oxygen or bone morphogenetic protein 7 (BMP7). Periosteal cells have been applied in bone tissue engineering; however, icariin effects on the proliferation and differentiation of periosteal cells is little reported.OBJECTIVE: To investigate the effect of icariin on the proliferation and differentiation of human periosteal cells, thus providing theoretical basis for icariin applied in bone tissue engineering. METHODS:The human periosteum was obtained and the primary cells were isolated in vitro.After culture and expansion,periosteal cells were cultured in 24-well plates, and induced by 0.001, 0.01 and 0.1 mg/L icariin and 50 μg/L BMP7, respectively. The corresponding avsorbance values of different groups were detected. The levels of alkaline phosphatase and calcium nodules in periosteal cells were measured at 1, 3, 5 and 7 days, and the mRNA levels of osteocalcin, osteopontin and Runx2 were detected at 3, 5 and 7 days. RESULTS AND CONCLUSION: The periosteal cells proliferated well after induction with icariin, and could proliferate well in different concentrations of icariin and the positive control group (P < 0.05). Compared with the control group, the periosteal cells induced by icariin were able to produce more alkaline phosphatase and calcium nodules (P < 0.05). The mRNA expression of osteocalcin, osteopontin and Runx2 in periosteal cells could be up-regulated by icariin (P < 0.05). These findings imply that icariin can promote proliferation and differentiate of periosteal cells into osteoblasts, and it can be used as an inducer for the preparation of seed cells in bone tissue engineering.

BACKGROUND: Although icariin has a clear role in inducing differentiation of bone marrow stromal cells, no studies have reported on its use in proliferation of periosteal cells in animal experiments. Three-dimensional polycaprolactone scaffold is a new scaffold, and it has the characteristics of biodegradability, proper pore size, and good histocompatibility. OBJECTIVE: To investigate the effect of icariin on the proliferation of periosteal cells, to construct periosteal cells and three-dimensional scaffold composite tissue-engineered bone and study its therapeutic effect on radical defects in rabbits, so as to provide a new way for the treatment of bone defects . METHODS: The mandibular periosteum was obtained from the rabbit and the primary cells were obtained by tissue explant method. The passage 3 cells were planted on 24-well plates. In experimental group, icariin at 10-4, 10-3and 10-2mol/L were added into the corresponding cell culture wells; equal volume of PBS was added to control group; positive control group was treated with 10 μg/L basic fibroblast growth factor. The cell proliferation in each group was measured by MTT method on the 2nd, 4th, 6thand 8thdays, respectively. The three-dimensional polycaprolactone scaffolds,10-2mol/L icariin and 107/L periosteum cells were placed in cell culture plates and cultured in vitro to construct the cell-scaffold complex. After the rabbit radical defect model was established, the model rabbits were randomized into three groups (n=15 per group), followed by implanted with scaffold complex (experimental group), polycaprolactone scaffold (polycaprolactone group) or nothing (control group). Pathological sections were taken at the 2nd, 4thand 8thweeks respectively for hematoxylin-eosin staining to observe the bone and count the number of osteoblasts. The X-ray films of the radius were obtained at the 4thweek. RESULTS AND CONCLUSION: (1) The proliferation of periosteal cells induced by different concentrations of icariin, especially 10-2mol/L icariin, was better than that in the control group (P < 0.05), Therefore, the optimal concentration of icariin was 10-2mol/L. (2) At 4 weeks after transplantation, in the experimental group, there were more osterocytes and less microvessels; in the polycaprolactone group, there were more new vessels, less woven newly born bone and osterocytes; in the control group, the defect region was filled with more granulation tissue and less osterocytes. At the 8thweek, complete healing in the defect region was observed in the cell-scaffold complex group, while partial healing in the polycaprolactone scaffold group. In the control group, however, fibroblasts and scar tissues were visible in the defect region, with presence of closed medullary cavity and nonunion. The number of osteocytes in the experimental group was significantly higher than that in the other two groups (P < 0.05). X-ray examination at the 4thweek showed that the defect region in the experimental group achieved healing;in the polycaprolactone group, the callus formed and defect region connected partly; and there was a clear gap in the defect region of the control group. Our findings indicate that icariin at different concentrations can promote the proliferation of periosteal cells and the concentration of 10-2mol/L has the greatest role. Icariin can induce proliferation of periosteal cells in the three-dimensional polycaprolactone scaffold, which contributes to bone repair.

PBL teaching method is a new mode of teaching which is originated from the West and implemented into China in recent years with an expectation that it would mainly develop the students’ self-learning ability,and enhance their skills of comprehensive thinking and solving actual problems.The author summarizes the practical experience of using PBL teaching methods in the theory teaching in Department of Medical Microbiology and Human Parasitology,China Medical University in the past three years,and then proved this method is very helpful to improving the students’integrated thinking by analysis of sample.At the same time the results also suggested that the students showed high enthusiasm in discussing the cases.By this way,the students showed great subjective intiative in their studies.

OBJECTIVETo establish the method of determining the quantity of hypericin in Hypericum perforatum and determine the quantity of the hypericin in defferent medicinal materials and asepsis seedings which grow in defferent environment.

METHODThe specimen is extracted with methanol--Pyridine (9:1) ultrasound extraction. Chromatographic assay is performed on a hypersily ODS2 (4.6 mm x 150 mm, 5 microm) column. The mobile phase is composed of methanol -1.56% dihydric natrium phosphate hydrogen natrium solution (shift solution's acidity to 2.1 with phosphoric acid)--ethyl acetate (4:1.9:1), velocity of flow is 1 mL x min(-1); column temperature is 35 degrees C; the detection wavelength is 590 nm.

RESULTA satisfactory seperaration between hypericin and impurity. The calibration curve is linear over the range of 0.0524-0.2620 microg for hypericin (r = 0.9998). The average recovery of hypericin is 97.50%.

CONCLUSIONThe quantity of hypericin in Hypericum perforatum has something to do with the genetic factor, environment factor, growing period and dry means. The method of determining the quantity of hypericin can be regarded as the method of controling the quantity of medicinal materials.

China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Hypericum ; chemistry ; genetics ; Perylene ; analogs & derivatives ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; genetics ; Quality Control ; Seasons

OBJECTIVETo evaluate the epidemiological status of HER2 protein expression in Chinese patients with gastric carcinoma, and to study its clinical and prognostic significance and the association with the clinicopathological features.

METHODSThe clinical data were reviewed in 860 patients with gastric carcinoma admitted to Guangdong General Hospital from 2003 to 2010. The HER2 status was evaluated using immunohistochemistry (IHC). The modified HercepTest scoring criterion was used to assess HER2 protein expression. The association between HER2 expression and clinicopathological features was analyzed by χ(2) test. Kaplan-Meier analysis, log-rank test and Cox regression model were used for the survival analysis.

RESULTSThe median age of the patients was 59 years, and the male-to-female ratio was 2.06:1. Positive expression of HER2 protein (3+) was found in 77 (9.0%) cases of gastric carcinoma, and in 69 (8.9%) advanced gastric cancers. There was significantly positive association between HER2 over-expression and tumor differentiation, Lauren classification and WHO classification. No significant association was observed between HER2 protein expression and patients' age, gender, tumor location and clinical stage. There was no statistically significant difference in survival rate between patients with positive HER2 expression and negative ones.

CONCLUSIONThough there was significantly positive association between HER2 expression status and tumor differentiation, histological type, it may be of limited prognostic value in gastric cancer patients.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; surgery ; Adenocarcinoma, Papillary ; metabolism ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Receptor, ErbB-2 ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Survival Rate ; Young Adult

OBJECTIVETo explore significance of high-risk human papillomavirus (HR-HPV) testing in atypical squamous cells, cannot exclude high grade squamous intraepithelial lesion (ASC-H).

METHODSPresence of HR-HPV DNA was examined in 45 patients with ASC-H using hybrid capture II (HC-II) test. Colposcopic examination and biopsy were taken all results were evaluated.

RESULTSOverall, 33 of 45 (73.3%) ASC-H cases were biopsy proven cervical intraepithelial lesion (CIN). 36 of 45 ASC-H cases were HPV-DNA positive, including 19 cases of HSIL and over lesion; whereas no HSIL or over was found in 9 HR-HPV negative cases. Sensitivity and negativity predictive value of HR-HPV in ASC-H with HSIL and over lesion were both 100%.

CONCLUSIONSASC-H strongly predicts the presence of HSIL, HR-HPV may serve as a predict select whether a patient with ASC-H should take colposcopic examination immediately, patients with positive HR-HPV should undergo immediate colposcopic examination, while negative HR-HPV is an excellent predictor of the absence of HSIL.

Adult ; Aged ; Biopsy ; Carcinoma, Squamous Cell ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Colposcopy ; DNA, Viral ; analysis ; Female ; Humans ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; Precancerous Conditions ; pathology ; virology ; Uterine Cervical Neoplasms ; pathology ; virology ; Uterine Cervicitis ; pathology ; virology ; Vaginal Smears ; Young Adult

OBJECTIVETo assess the sensitivity and positive predictive value (PPV) of intraoperative frozen section diagnosis of the borderline tumor of ovary (BTO).

METHODSA retrospective analysis and comparison were done respectively between the accuracies of diagnoses made by using frozen and paraffin sections from the same tissue blocks for BTO from March 1995 to May 2008 achieved in the Department of Pathology, Guangdong General Hospital. Univariate and multivariate regression models were used to assess the influence of patient and tumor characteristics on the likelihood of underdiagnosis and overdiagnosis.

RESULTSOf the 73 patients analyzed, 39 cases (53.42%) were histologically serous tumors, 32 (43.84%) were mucinous and 2 (2.74%) were endometrioid tumors. Diagnoses identical in those made by using either frozen or routine paraffin sections were 55/73 (75.34%). The sensitivity and positive predictive value of frozen section diagnosis were 87.30% and 85.94%, respectively. Underdiagnosis of frozen section were 18/73 (24.66%). There was no overdiagnosis cases obtained. Univariate analysis showed that tumor diameter and tumor histology were the predictors of underdiagnosis in frozen section analysis. And in multivariate analysis, only tumor diameter, rather than patient age, tumor histology and stage, bilateral side tumor, serum CA-125 and concurrent presence of endometriosis was a predictor of underdiagnosis.

CONCLUSIONSIntraoperative frozen section diagnosis of BTO has a low sensitivity and PPV. Underdiagnosis is not uncommon. Surgical management based on intraoperative frozen section diagnosis should be used with caution.

Adult ; Aged ; Aged, 80 and over ; CA-125 Antigen ; metabolism ; Carcinoma, Endometrioid ; metabolism ; pathology ; surgery ; Cystadenoma, Mucinous ; metabolism ; pathology ; surgery ; Cystadenoma, Serous ; metabolism ; pathology ; surgery ; Female ; Frozen Sections ; Humans ; Intraoperative Period ; Middle Aged ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; surgery ; Paraffin Embedding ; Predictive Value of Tests ; Retrospective Studies ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo investigate the outcome of the children with thoracic spine tuberculosis who underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach.

METHODSFrom June 2005 to December 2010,9 children with thoracic spine tuberculosis underwent radical debridement, reconstruction with bone autograft or allograft and internal fixation via posterior approach including 7 males and 2 females with an average age of 7 years old ranging from 3 to 12 years. The course of the disease ranged from 3 months to 1 year (averaged 6 months). All the patients had continuous thoracic and back pain,intercostal nerve pain and kyphosis deformity, accompanied with low fever,night sweat and pathologic leanness. The predilection sites examined by X-ray, CT or MRI were in T4 to T9 segment. The kyphosis angle ranged from 35 degrees to 72 degrees (averaged 48.2 degrees) before surgery. The ASIA classification was as follows: 2 cases at grade B, 5 at grade C, 2 at grade D. All the patients underwent a standard one-stage operation via posterior approach. Radical debridement was performed, then iliac crest bone autograft or allograft was placed and transpedicular screw system internal fixation was done to reconstruct the spinal column. The change of kyphosis angle and fusion of bone grafting were reexamined by X-ray regularly. The neurological function were evaluated according to ASIA classification.

RESULTSThere was no injury of blood vessel or spinal cord during the surgery. Nine children were followed-up for 16 to 38 months (averaged 24 months). The tuberculosis symptoms disappeared after surgery and there was no tuberculosis recurrence,incision infection, sinus formation and internal fixation failure in any of these chiildren. ESR reexamination recovered normally. Bony fusion was obtained in all patients and internal fixation position was normal 4 to 8 month postoperatively. The kyphosis angle ranged from 12 degrees to 30 degrees (averaged 19.50) at final followed-up. The function of spinal cord improved postoperatively, the function of spinal cord recovered at different degrees: 2 cases at grade C, 2 at grade D, and 5 at grade E.

CONCLUSIONThe one-stage posterior approach can provide direct and safe access to the lesion. The effect of vertebral canal decompression and kyphosis deformity correction were significantly. The structural iliac crest autograft or allograft and posterior transpedicular screw system could work effectively to stabilize the thoracic junction.

Bone Transplantation ; Child ; Child, Preschool ; Debridement ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; surgery

OBJECTIVETo examine the activation and cytotoxicity of human peripheral blood T lymphocyte induced in vitro by human 4-1-BB ligand (4-1-BBL) gene transfected into tumor Tca8113 cells.

METHODSThe eukaryotic expression vector pEGFP-h4-1-BBL was transfected into human oral carcinoma cell line Tca8113 by Lipofectamine 2000. The transfected cells were then selected in medium containing G-418, cloned by limited dilution and named as Tca8113-4-1-BBL. Human 4-1-BBL mRNA and protein expression of transfected cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting respectively. The tumor cell vaccines (TCV) were obtained by treatment with mitomycin (MMC). Human peripheral blood mononuclear cells (PBMC) were prepared from lymphoprep, and then stimulated with anti-CD-3 mAb and incubated with non-transfected or transfected TCV-Tca8113 cells, respectively. The proliferation of T cells was evaluated by trypan blue exclusion; the CCK-8 was used to detect the cytotoxic effect of T lymphocytes. Meanwhile, the secretion of interferon-gamma (IFN-gamma) and interleukin (IL)-2 in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe Tca8113 cells transfected by pEGFP-h4-1-BBL could express human 4-1-BBL efficiently. As compared with wild type Tca8113 cells, the transfected Tca8113 cells could markedly promote proliferation, IL-2 and IFN-gamma production and cytotoxic activity of lymphocytes.

CONCLUSIONSThe transfection of human 4-1-BBL gene in Tca8113 cells is effective in enhancing its immunogenicity and inducing antitumor immune response in vitro.

4-1BB Ligand ; genetics ; Carcinoma, Squamous Cell ; genetics ; immunology ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Interferon-gamma ; immunology ; Interleukin-2 ; immunology ; Lymphocyte Activation ; Mouth Neoplasms ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection