Objective The aim of our present study is to culture a new female lung adenocarcinoma cell line in Xuanwei region of Yunnan province,detect its proliferation dynamics and heterogeneity transplant and provide an ideal experimental model for the study of lung cancer.Methods The cell line was derived from a surgical specimen of lung cancer,and obtained lung adenocarcinoma cell clone by limiting dilution.Its biologic characteristics were explored by light microscope,determination of the doubling time and growth curve,inoculation the cells in mice as well as immunohistochemistry.Results Morphological study,proliferation dynamics and immersed growth demonstrated the cultured cells' malignant characteristics.The cell line produced 100% tumors after axillary transplantation into the mice provided by the Animal Centre of Kunming Medical College.The results of immunohistochemistry indicated the cells' adenocarcinoma characteristics.Conclusions According to a series of analysis of the cell line in vitro,the present study gave an important implication and some theory basis of following research on human lung adenocarcinoma,biological characteristics,cancerization,the mechanism of metastasis,and exploitation for new anticancer medicine et al.

Objective To evaluate anti-HEV IgG and IgM diagnostic kits with sera from convalescent hepatitis E patients and to establish the quantification method of detecting anti-HEV lgG.Methods Detect 42 convalescent serum samples of over 6 months after onset of hepatitis E patients from Jiangsu province with anti-HEV IgM and IgG diagnostic kits. Select and mix the anti-HEV IgG positive sera which were confirmed by Western blot with ORF2 and ORF3 antigen. The mixed serum was calibrated with a WHO anti-HEV Ig standard. A series quantitative linear standard was made for quantitative detection of anti-HEV IgG in hepatitis E vaccine clinical trials phase Ⅲ. Results The positive rates of the anti-HEV IgG di-agnose kits of G, K, MP, Wantai were 71.4%, 78.6%, 92.9% and 100% respectively. The positive rates of G was lower than that of MP (χ~2 = 5.19, P<0.05) and obviously lower than Wantai (χ~2 = 11.76,P<0.01). The positive rates of K was also obviously lower than that of Wantai (χ~2 =7.96, P <0.01).The positive rates of the anti-HEV IgM diagnose kits of MP, G, X, Wantai, K were 21.4%, 7.1%,21.4%, 64.3%, 78.6% respectively. The positive rate of both K and Wantai were obviously higher than that of MP(χ~2 = 15.75 ,P<0.01 ; X2 = 27.43 ,P< 0.01). With the Western blot confirmation test, 30 and 18 sera were reactive to ORF2 and ORF3 antigen separately. The anti-HEV IgG concentration of HEV-D01 mixed by 13 samples was 57.94 U/ml by the calibration. Prepare seven 1.5-fold dilution series of quantita-tive linear standard for HEV vaccine clinical trials phase Ⅲ, concentration range from 0.077 to 0.877 U/ml. The quantitive values of high, medium and low concentrations quality control samples lay in the range of average ± 2s, and the CV of quantitative values were 16%, 16%, 12% respectively. Conclusion The quality of different anti-HEY IgM and IgG diagnose kits were different. This study had set up a set of anti-HEV IgG linear quantitative standard, which fit for detecting anti-HEV IgG antibodies quantitatively in HEVvaccine clinical trial phase Ⅲ.

Objective To study the clinical features,pathological characters,treatment and prognosis of patients with malignant solitary fibrous tumor (MSFT).Methods Retrospective analysis was made on the clinical data of 11 MSFT patients undergoing surgical resection in Beijing Shijitan Hospital from Jan 2010 to Dec 2017.Overall survival (OS) and disease-free survival (DFS) were assessed using Kaplan-Meier methods.Results One patient died of duodenal fistula within a month after surgery,and the other 10 patients recovered with no severe complications.Immunohistochemically,the tumor cells are all positive for CD34 (11/11) and vimentin (10/10) in a different degree.The median OS and DFS for the 10 patients were 49 and 26 months respectively.The 1-,3-,and 5-year OS rates were 89％,56％,42％ respectively.The 1-,3-,and 5-year DFS rates were 79％,34％,23％ respectively.Conclusion MSFT is a malignancy with high recurrence rate.Tumor recurrence was the main cause of death for patients with MSFT.

Objective: To analyze the clinicopathological characteristics of ovarian pseudomyxoma peritonei (PMP). Methods: Clinical and pathological data from a total of 272 PMP patients diagnosed at Beijing Shijitan Hospital from January 2010 to January 2019 were collected from a database and retrospectively analyzed to study the origin of PMP tumors. Pathological slides marked with antigens were further studied using immunohistochemical staining, including CK7, CK20, CEA, Villin, CDX2, SATB2, CA125, ER, PR, MUC1, and MUC2. Results: Among the 272 PMP patients studied, the tumors of 245 (90.1%) originated from the appendix, while the remaining 27 (9.9%) originated from non-appendix tissues, including 5 (1.8%) from the ovaries. Ovarian cases included two ovarian teratomas, two ovarian mucinous cystadenomas, and one borderline ovarian mucinous cystadenoma. Histopathological analysis of peritoneal me-tastases further revealed two acellular mucins, two low-grade mucinous carcinoma peritonei, and one high-grade mucinous carcinoma peritonei, while immunohistochemistry revealed positive staining for CK20, CEA, Villin, and CDX2; SATB2 was also found to be partially positive in teratomas with mucinous tumors: negative in two cases and partially positive in one case. Conclusions: The occurrence of ovarian PMP is rare. Its precise diagnosis demands for a serial section of the whole appendix or suspected tissue to exclude any appen-diceal mucinous neoplasms, as well as the combination of a comprehensive analysis of its clinical signs and symptoms, imaging find-ings, surgical findings, histopathological characteristics, and immunohistochemistry.

OBJECTIVETo evaluate the corresponding influence on pulmonary embolism incidence between immobilization and exercise in different stage of thrombus after acute deep vein thrombosis in rabbits.

METHODSForty-eight New Zealand rabbits were randomly divided into three groups depending on the different organized stage of thrombus: the early, medium and later stage group.Each group was subdivided into two sub groups: the immobile and mobile subgroup. Rabbit modeling of deep vein thrombosis was made by ligating the right femoral vein. Among the early-stage group, rabbits of the immobile subgroup were fixed for 3 days, while that of the mobile subgroup were free to move for 3 days, then each was euthanized to extract the lungs for pathological examination. Among the medium-stage group, each of the immobile subgroup were fixed for 7 days, while the mobile subgroup ones were fixed for 3 days, then released free-moving for 4 days following the pathological extraction. Among the later-stage group, animals in the immobile subgroup were fixed for 14 days comparing the mobile subgroup fixed for 7 days and next free-moving for 7 days, then each was euthanized.

RESULTSAmong the early-stage group, pulmonary embolism incidence (PEI) of the immobile and mobile subgroup was 4/8 vs.3/8, the pulmonary lobe embolism incidence (PLEI) was 17.5% (7/40) vs. 15.0% (6/40). Among the medium-stage group, PEI of the immobile and mobile subgroup was 3/8 vs. 2/8, PLEI was 37.5% (7/40) vs. 25.0% (10/40). Among the later-stage group, PEI of the immobile and mobile subgroup was 3/8 vs. 3/8, PLEI was 12.5% (5/40) vs. 15.0% (6/40). There was no statistical difference between immobilization subgroup and mobilization subgroup among different stage group.

CONCLUSIONOn the premise of given anticoagulation treatment, early ambulation do not significantly increase pulmonary embolism incidence after acute deep vein thrombosis of lower extremity in rabbits.

Animals ; Disease Models, Animal ; Immobilization ; Lung ; pathology ; Motor Activity ; Pulmonary Embolism ; etiology ; Rabbits ; Time Factors ; Venous Thrombosis ; complications

Objective:s To evaluate the impacts of prior surgical scores(PSS) on the clinical efficacy and perioperative safety of cytoreductive surgery(CRS) and hyperthermic intraperitoneal chemotherapy(HIPEC) for pseudomyxoma peritonei(PMP).Methods:From the comprehensive PMP database, we collect the cases treated for the first time by CRS+ HIPEC, to form this study cohort. The clinicopathological features, PSS, CRS+ HIPEC details, overall survival(OS), and serious adverse events(SAEs) are systematically analyzed, to study the correlations between PSS and OS or SAEs.Results:335 PMP cases received standardized CRS+ HIPEC in this study. The median OS is 58.2 months for PSS-0 patients, 63.7 months for PSS-1, and 55.4 months for PSS-2/3, with no statistically significant differences in OS among the different PSS groups(χ 2=0.499, P=0.779). Subgroup analysis by pathologic types also found no statistically significant differences among the different PSS groups. Moreover, no significantly statistical differences are observed in overall SAEs(χ 2=0.625, P=0.722), CRS-related SAEs(χ 2=0.267, P=0.901), and non-CRS-related SAEs(χ 2=0.677, P=0.715), among the different PSS groups. Conclusions:PSS does not pose significant impacts on the efficacy and safety of CRS+ HIPEC for PMP patients at experienced treatment center.

Objective: To analyze the pathological features of pseudomyxoma peritonei (PMP) in correlation with the survival status and independent prognostic factors. Methods: One-hundred and fifty-five PMP specimens were collected at Beijing Shijitan Hospital, Capital Medical University, from 2012 to 2018. Conventional histopathological evaluation was performed to document the primary tumor site, histopathological type, lymph nodes metastasis, tumor emboli in the blood and lymph vessels, nerve invasion and cellular density. The immunohistochemical parameters including Ki-67, p53, MMR-related protein, MUC2 and MUC5AC were analyzed. Clinical follow-up data were reviewed to correlate with pathological prognostic factors using Kaplan-Meier estimator and Cox proportional hazards regression model for univariate and multivariate analysis. Results: Among 155 PMP patients, there were 77 males and 78 females. There were 98 cases (63.2%) of low-grade peritoneal mucinous carcinomatosis, 49 cases (31.6%) of high-grade peritoneal mucinous carcinomatosis, 8 cases (5.2%) of high-grade mucinous carcinoma peritonei with signet ring cells; only 15 cases (9.7%) with lymph node metastasis; 18 cases (11.6%) with tumor emboli in the blood and lymph vessels; 8/126 (6.3%) were positive dMMR; 100 cases (64.5%) had Ki-67 label index <50%, and 56 cases(36.1%) presented with mutant type p53. Univariate analysis revealed 11 survival-related pathological parameters including gender, age, primary tumor site, histopathological type, lymph node metastasis, tumor emboli in the blood and lymph vessels, nerve invasion, cellular density, Ki-67 label index rate, p53 and dMMR. Multivariate analysis identified 4 independent prognostic factors including the histopathological type (HR 59.78, P<0.01), lymph node metastasis (HR 3.74, P=0.028), nerve invasion (HR 7.81, P=0.007) and dMMR (HR 9.82, P<0.01). Conclusions Histopathological type is the most important prognostic factor of PMP with dMMR as an independent molecular prognostic indicator.

Objective: To establish the patient derived xenograft (PDX) model of pseudomyxoma peritonei (PMP), and identify the key characteristics of tumor biology of this model, in order to provide a reliable model for studying the pathological mechanisms and new therapeutic strategies of PMP. Methods: PMP tumor tissue was obtained from surgery and cut into pieces after washing. Then tumor pieces were implanted subcutaneously in BAL B/c-nu mice for 6 stable passages. In the 7th passage, tumor tissue was implanted orthotopically into abdomen. Subcutaneous tumor and orthotopic tumor were then homogenized to make tumor cell suspension, implanted into abdomen of 10 BAL B/c-nu mice through midline laparotomy, 100 μl for each. The key experimental parameters including body weight changes in the observation period, experimental peritoneal cancer index (ePCI) score at the autopsy, histopathological and immunohistochemical characteristics, and gene expression profiles by high-throughput whole-genome exon sequencing were detected and recorded. Results: The successful rate of established orthotopic PDX model of human PMP was 100% (10/10). The animals showed smooth body weight increases after tumor inoculation until day 27, then the body weight began to decrease steadily. Widespread tumor dissemination of PMP tumor through the whole abdomen was found by autopsy, including the diaphragm, liver, spleen, stomach, kidney, parietal peritoneum, bowel and mesenterium. Gelatinous ascites was also observed in abdominopelvic cavity. The ePCI score ranged from 5 to 9, with a 8 of median ePCI. Histopathological studies showed peritoneal mucinous carcinomatosis accompanied with signet ring cells (PMCA-S), obvious tumor cell atypia and parenchymal invasion.Immunohistochemistry showed the expressions of MUC1, MUC2, MUC5AC, CEA, CA199, CK20, CDX-2 and Ki-67 were positive, MUC6, CK7 and p53 were negative. Whole-exome sequencing identified that the most significant genetic alteration is the exon10 missense mutation c. 1621A>C of KIT gene, the mutation abundance was 89.7%. Conclusion PDX model of PMCA-S is successfully established, which displays the characters of high-degree malignancy, high proliferation and strong aggressiveness.

Objective: To explore the relationship between liver controlled attenuation parameters (CAP) and body fat mass and its distribution. Methods: From May to December 2018, 978 adult patients visited at the fatty liver center of the Third People's Hospital of Changzhou were treated. The patient's liver controlled attenuation parameters were measured by transient elastography and the body fat mass and its distribution were measured by bioelectrical impedance technology. Pearson’s correlation coefficient was adopted to describe the correlation between liver CAP value and body mass index (BMI), body fat mass index (BFMI), trunk fat mass index (TFMI), limbs fat mass index (LFMI) and visceral fat area (VFA). Receiver operating characteristic curve (ROC) and area under the curve (AUC) were used to evaluate BMI, BFMI, TFMI, LFMI and VFA to differentiate the cut-off points and efficacy of CAP for diagnosing grading of fatty liver changes in S0-1 and S2-3. Results: In 653 cases of male, S0 ~ S3 accounted for 4.90%, 3.37%, 22.36% and 69.37%, respectively, and in 325 cases of females, S0 ~ S3 accounted for 7.38%, 6.46%, 13.23% and 72.92%, respectively. Female patients had more visceral, trunk and limbs fat than male (P < 0.01). Body mass, body fat mass, body fat percentage, BMI, BFMI, TFMI, LFMI, and VFA were increased in male and female patients with increasing liver fat grade (P < 0.01). CAP values ​​of male and female patients were positively correlated with BMI, BFMI, TFMI, LFMI and VFA. Percentage of body fat mass increased with increasing liver fat grade (male: F = 13.42, P < 0.001; female: F = 3.22, P = 0.023); while limb fat mass percentage did not increase with liver fat grade (Male: F = 1.13, P = 0.34; female: F = 1.05, P = 0.37). Hepatic steatosis grading (S0 ~ 1 or S2 ~ 3) diagnosed with CAP were distinguished through BMI, BFMI, TFMI, LFMI and VFA. AUC was 0.80 ~ 0.82 in males (P < 0.01), and 0.75 ~ 0.78 in females (P < 0.01). Conclusion The liver CAP value is positively correlated with the body's limbs, trunk and visceral fat, and has a strong correlation with trunk and visceral fat. BMI, BFMI, TFMI, LFMI and VFA up to some extent can identify the CAP diagnosis of grading of fatty liver changes in S0-1 and S2-3.

BACKGROUND: The new emerging avian influenza A H7N9 virus, causing severe human infection with a mortality rate of around 41%. This study aims to provide a novel treatment option for the prevention and control of H7N9. METHODS: H7 hemagglutinin (HA)-specific B cells were isolated from peripheral blood plasma cells of the patients previously infected by H7N9 in Jiangsu Province, China. The human monoclonal antibodies (mAbs) were generated by amplification and cloning of these HA-specific B cells. First, all human mAbs were screened for binding activity by enzyme-linked immunosorbent assay. Then, those mAbs, exhibiting potent affinity to recognize H7 HAs were further evaluated by hemagglutination-inhibiting (HAI) and microneutralization in vitro assays. Finally, the lead mAb candidate was selected and tested against the lethal challenge of the H7N9 virus using murine models. RESULTS: The mAb 6-137 was able to recognize a panel of H7 HAs with high affinity but not HA of other subtypes, including H1N1 and H3N2. The mAb 6-137 can efficiently inhibit the HA activity in the inactivated H7N9 virus and neutralize 100 tissue culture infectious dose 50 (TCID50) of H7N9 virus (influenza A/Nanjing/1/2013) in vitro, with neutralizing activity as low as 78 ng/mL. In addition, the mAb 6-137 protected the mice against the lethal challenge of H7N9 prophylactically and therapeutically. CONCLUSION The mAb 6-137 could be an effective antibody as a prophylactic or therapeutic biological treatment for the H7N9 exposure or infection.
Animals ; Antibodies, Monoclonal/therapeutic use* ; Antibodies, Neutralizing/therapeutic use* ; Antibodies, Viral ; Hemagglutinins ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza A Virus, H3N2 Subtype ; Influenza A Virus, H7N9 Subtype ; Influenza Vaccines ; Influenza in Birds ; Influenza, Human/prevention & control* ; Mice