Objective To remove cellular compenents from porcine aortic valve with different reagents, providing acellular tissue matrix(ACTM) scaffolds for tissue engineering of heart valve. Methods Different detergents (TritonX-100、Sodium dodecyl sulfate and sodium deoxycholate) and enzyme (trypsin) were used to remove cells and its, compenents from porcine aortic valves, respectively. According to the different detergents, specimens of porcine aortic valve were divided into three groups(Sodium dodecyl sulfate, sodium deoxycholate, and Triton X-100). Specimens were observed grossly uncroscopically. Haematoxylin-eosin and DNA assay was performed to confirm the removal of cells. Transmission electron microscope was used to observe the integrity of collagen and elastin. The changes of mechanical properties were also studied. Results The cells were removed effectively from cusps and roots of porcine aortic valves by Triton X-100 and sodium dodecyl sulfate in 96 h. Sodium deoxycholate could not effectively remove the cells from the root of aortic valve. Treatment with SDS disrupted collagen fiber structure of porcine aortic valve, resulting in poor mechanical properties. Treatment with TritonX-100 result in an acellular porcine aortic valve matrix with retention of near normal structure and mechanical properties. Conclusion Porcine aortic valve can be successfully decellularised with retention of near normal structure and mechanical properties by Triton X-100.

OBJECTIVE: To study the function of Qiyeling Decoction in inducing apoptosis of transplanted human lung adenocarcinoma cells A549 in nude mice. METHODS: Nude mice with transplanted A549 tumor were randomly divided into the untreated control group (group A), chemotherapy treated group (group B), chemotherapy plus Qiyeling Decoction treated group (group C), Qiyeling Decoction treated group (group D) and managed correspondingly. The tumor volume was measured and calculated into tumor weight. The apoptosis of tumor cells were examined using in situ cell apoptosis detection kit. RESULTS: The tumor weight was lower obviously in groups B, C and D than that in group A (P<0.05). The apoptosis of tumor cells was lower obviously in groups B, and C than that in group D (P<0.05). Cells in group A appeared perfect differentiation during the early stage and apoptosis later. CONCLUSION: Qiyeling Decoction can induce A549 cell apoptosis in nude mice.

OBJECTIVETo evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC).

METHODSThe identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit.

RESULTSThe lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells.

CONCLUSIONThe plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; blood ; pathology ; DNA-Binding Proteins ; Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Lung Neoplasms ; blood ; pathology ; Lymphokines ; blood ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplastic Cells, Circulating ; chemistry ; pathology ; Telomerase ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective: To investigation the situation of cold chain on vaccine in parts of Zhejiang Province and to provide recommendations for the management. Methods: From October to December, 2016, we each selected an immunization clinic in Cangnan County of Wenzhou, Yongkang City of Jinhua, Jianggan District of Hangzhou. Temperature recorder and vaccine viral monitor (VVM) labels were used to monitor the cold chain during all the storage and transportation process. In Jianggan District, we use optical density sensor to detected 20 VVM labels every time when the vaccine was stock in and out. Results: In total, 54 958 records were collected by temperature recording devices in all the three immunization clinic. 275 records exceeded the temperature limit required for store and transportation, of which 270 (98.2%) were above 8 ℃ and 5 (1.9%) were under 2 ℃. Excessive temperature exposure mainly occurred during the transportation (38.2%, n=105), followed by storage process in CDCs at different levels (26.2%, n=72), stock in and out (20.7%, n=50) and storage in the refrigerators in immunization clinics (14.9%, n=41). The average optical density difference between VVM labels and the reference circular decreased from 0.404 to 0.344 when the vaccines were delivered from the Zhejiang provincial CDC to immunization clinics. The color of VVMs did not significantly changed before use. Conclusions The potential risk of vaccine cold chain in the monitoring sites is over-temperature. The weak links of cold chain management include the transportation, storage process, and stock in and out.

Objective: To investigate the clinical and pathological characteristics and prognosis of ovarian clear cell borderline tumor. Methods: A total of 12 cases of ovarian clear cell borderline tumors recorded were collected from May 2011 to December 2017 at Obstetrics and Gynecology Hospital, Fudan University.Clinical histories were retrieved and pathological slides were reviewed. Results: The age of the patients ranged from 35 to 65 years with a mean age of 52 years. Seven cases were associated with cystic endometriosis of the ovary. All tumors consisted of irregular and crowded glands or cysts embedded in a fibromatous stroma. The cysts and glands were lined by mild to moderate atypical cells.CK7 and HNF-1β were expressed in all cases, and Naspin A was expressed in 11 cases. ARID1A expression was absent in 5 cases and p53 showed wild-type expression. None of the cases developed recurrence during follow-up ranging from 7 to 79 months. Conclusions Ovarian clear cell borderline tumor may be associated with endometriosis and tumor suppressor gene ARIDA. The tumor has a good prognosis without recurrence and progression to carcinoma.

Objective: To describe the clinicopathological features of synchronous mucinous metaplasia and neoplasia of the female genital tract (SMMN-FGT). Methods: The sample consisted of 7 cases of SMMN-FGT recorded from November 2014 to September 2017 at Obstetrics and Gynecology Hospital, Fudan University.PAP method was used in immunohistochemistry.Clinical histories were retrieved and pathological slides were reviewed. Results: The patients were 37 to 70 years old(mean 54 years old). All patients showed endometrial mucinous lesions associated with cervical lesions. Three cases were an admixture of minimal deviation adenocarcinoma(MDA) and gastrictype adenocarcinoma(GAS). Three cases were an admixture of lobular endocervical glandular hyperplasia (LEGH), atypical LEGH and focal gastrictype adenocarcinoma in situ, one of which had early invasive gastrictype adenocarcinoma.Endometrium showed a structure of LEGH in one patient with focal simple gastric mucinous metaplasia in her cervix. Gastric mucinous differentiation was found in unilateral fallopian tube in 6 patients. Ovarian mucinous lesions were found in 3 patients. p16 was negative staining in 6 cases and positive in 1 case. CK7 was diffusely positive in all lesions. CK20 and CDX2 were negative or only focally positive.The expression of MUC6 was strongly positive staining or focal staining. p53 in GAS and GAS in situ had mutant expression, but wild expression in MDA region. Patients were followed up for 2 to 34 months and no recurrence was found. Conclusions SMMN-FGT is a series of rare mucinous lesions involving multiple areas of the female genital tract, including benign or malignant lesions with gastric differentiation. It is not related to infection with high-risk human papilloma virus. When cervical gastrictype lesions are found, SMMN-FGT should be considered and should be differentiated from metastatic mucinous adenocarcinoma.

Objective To investigate the relationship between the expression of CENPF in NSCLC adenocarcinoma (LUAD) and the clinical prognosis of patients and its effect on the metastasis of lung adenocarcinoma cells. Methods The expression of CENPF in LUAD and its relationship with patient prognosis were analyzed by online bioinformatics. The expression of CENPF was verified by LUAD tissue microarray immunohistochemical staining. Kaplan-Meier analysis was performed to analyze the relationship between the expression of CENPF and the prognosis of patients with lung adenocarcinoma. Cox survival hazard ratio was used to analyze the factors affecting the survival of patients. Chi-square analysis was adopted to examine the relationship between CENPF expression and clinicopathological stage and grade of patients. The expression of CENPF in NCI-H2126 cells were knocked out by lentivirus, and then the proliferation, invasion, and migration abilities of the cells were detected. Changes in mRNA expression profiles after CENPF knockout were detected by RNA-seq. Bioinformatics analysis of downstream signaling pathways and the target genes of CENPF was also performed. Western blot was used to verify the target gene. Results CENPF was significantly upregulated in LUAD tumor tissue (P < 0.05) and significantly correlated with pathological stage (P=0.013). The higher expression of CENPF, the worse the prognosis of patients (P=0.01, P=0.027). After the expression was CENPF of knocked out, the cell proliferation, migration, and invasion abilities significantly reduced (P < 0.01). The expression of chemokine pathway genes in cells was enriched significantly (P < 0.001). ACKR3/CXCR7 and CDH2/N-cadherin were significantly downregulated, whereas CDH1/E-cadherin was significantly upregulated. After CENPF was knocked out, ACKR3/CXCR7 and N-cadherin were significantly downregulated, whereas E-cadherin significantly increased. Conclusion The expression of CENPF is negatively correlated with the clinical prognosis of patients with LUAD, and it promotes the occurrence of EMT by regulating the expression levels of N-cadherin and E-cadherin related to EMT through ACKR3/CXCR7.

Objective:To investigate the clinical and pathological characteristics of ovarian cellular fibromas.Methods:The sample con-sisted of 24 cases of ovarian cellular fibromas from February 2008 to March 2017 in the Obstetrics and Gynecology Hospital of Fudan University.Clinical histories were retrieved,and pathological slides were reviewed.Results:The age of the patients ranged from 17 to 70 years old,with a mean age of 46.5 years.Clinical symptoms included ovarian masses,abdominal pain,or pleuroperitoneal fluid.Se-rum CA125 notably increased in two patients.Of the 24 cases,13 and 10 occurred in the right and left ovaries,respectively,and one case occurred bilaterally.Tumor cells were densely cellular in all cases and were mitotically active in three cases(5-7/10 high power fields).A minor component of sex cord elements(<10% area of the tumor)was present in three cases,and luteinized cells were ob-served in four cases.None of the cases manifested recurrence during follow-up ranging from 1 month to 109 months.Conclusion:Ovarian cellular fibromas are pure ovarian stromal tumors that may manifest mitotic activity, sex cord elements, and luteinization. Some patients present with pleuroperitoneal fluid and increased CA125.Thus,the pathological features of cellular fibromas must be mastered to avoid misdiagnosis for other benign or malignant tumors and improper treatment.

Adenocarcinoma, Clear Cell ; diagnosis ; Biomarkers, Tumor ; Female ; Humans ; Ovarian Neoplasms ; diagnosis ; Prognosis