Objective To establish an animal model of hepatic inflow occlusion with meso-caval shunt in Bama swines so as to evaluate the tolerance limit of normothermic hepatic inflow occlusion.Methods Eighteen Bama swines were divided into group A in which the hepatic blood inflow was occluded for 90 min and group B in which the hepatic blood inflow was blocked for 100 min.The animal survival rate,the changes of liver function and histopathology were observed.Results The animal survival rate was significantly higher in group A than group B(100% vs 44.4%,P

Carcinoma, Squamous Cell ; Epithelial Cells ; Humans ; Papilloma

Objective To explore the role of portal venous pressure changes in the liver dysfunction caused by hepatic congestion after extended liver resection.Methods The experimental study was adopted.According to the random number table,90 Sprague-Dawley rats were divided into 3 groups,30 in each group:30 rats in the non-congestion group received 70％ of liver resection (median lobe + left lobe),30 rats in the congestion group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and 30 rats in the congestion + splenectomy group received 70％ of liver resection (median lobe + left lobe) with whole caudal lobe congestion by ligation of veins and splenectomy.(1) Twenty rats in each group were used to make postoperative survival analysis.Ten rats in each group were used for related experiments.The portal venous pressures (PVPs) of 5 rats in each group were detected at postoperative 12 hours and 24 hours,and then blood and liver specimens were collected.(2) PVP changes were detected at postoperative 12 hours and 24 hours.(3) Clinical and biochemical test:level of total bilirubin (TBil) was tested at postoperative 12 hours and 24 hours.(4) Pathological examination:liver pathological damage was detected by HE staining.(5) The expression of CD68 macrophagocyte was detected by immunohistochemical staining.(6) The relative expressions of Cleaved Casepase-3 and hypoxia inducible factor-1α (HIF-1α) proteins at postoperative 24 hours were detected by Westein blot.(7) The relative expressions of mRNA of vascular regulation related genes (ET-1/eNOS) and inflammatory factors (TNF-α and IL-6) were detected by real-time polymerase chain reaction (RT-PCR).(8)The hyaluronic acid (HA) was measured by enzyme-linked immuno-sorbent assay (ELISA).Measurement data with normal distribution were represented as （x） ± s.Comparison among 3 groups was done using the ANOVA,and pairwise comparison was done by the LSD test.The postoperative 5-day survival curve was drawn by the KaplanMeier method,and the survival was compared using the Log-rank test.Results (1) Survival analysis:5-day survival rate in the non-congestion group,congestion group and congestion + splenectomy group were respectively 75％,10％ and 55％,with a statistically significant difference among the 3 groups (x2=18.21,P ＜0.05).(2)Changes of PVPs and TBil:levels of PVP and TBil in the non-congestion group,congestion group and congestion + splenectomy group were respectively (15.77 ±0.67)cmH2O,(18.33 ±0.28) cmH2O,(14.87 ± 0.58) cmH2O,(1.48 ±0.10)μmol/L,(1.76±0.15) μ mol/L,(1.62 ±0.11) μmol/L at postoperative 12 hours and (13.49 ± 0.45) cmH2 O,(16.96 ± 0.82) cmH2 O,(15.69 ± 0.85) cmH2 O,(1.47 ± 0.11) μmol/L,(1.94 ± 0.07) μmol/L,(1.67 ± 0.11) μmol/L at postoperative 24 hours,showing statistically significant differences among 3 groups (F =56.53,29.01,6.81,27.85,P ＜ 0.05).(3) Results of pathological examination:compared with noncongestion group,there were a lot of vacuolar cells with degeneration appearing in non-congestion liver tissues,severe liver cell swelling and hepatic sinus congestion in the congestion group at postoperative 24 hours.Compared with congestion group,vacuolar degeneration appearing in non-congestion liver tissues have some improvement in the congestion + splenectomy group.(4) Immunohistochemical staining:compared with non-congestion group and congestion + splenectomy group,the positive CD68 marked macrophages in the congestion group were increased at postoperative 24 hours.(5) Western blot assay:the relative expressions of Cleaved Casepase-3 and HIF-1α proteins in the non-congestion group,congestion group and congestion + splenectomy group were 0.63 ± 0.05,1.17 ± O.18,0.95 ± 0.17 and 0.63 ± 0.14,1.48 ± 0.08,1.13 ± 0.17,respectively,showing statistically significant differences among 3 groups (F =17.42,50.58,P ＜ 0.05).(6) Results of RT-PCR:the relative expression of mRNA of ET-1/eNOS in the non-congestion group,congestion group and congestion + splenectomy group was respectively 1.01 ± 0.63,2.09 ± 0.27,0.82 ± 0.12 at postoperative 12 hours and 0.73 ± 0.17,2.16 ± 0.94,0.80 ± 0.24 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =62.91,10.65,P ＜0.05).The relative expression of mRNA of TNF-α in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.99 ± 0.08,127.80 ± 13.15,7.34 ± 1.56 at postoperative 12 hours and 0.99 ± 0.06,116.62 ± 13.32,58.62 ± 12.12 at postoperative 24 hours,showing statistically significant differences among 3 groups (F =436.77,154.54,P ＜ 0.05).The relative expression of mRNA of IL-6 in the non-congestion group,congestion group and congestion + splenectomy group was respectively 0.98 ±0.06,1.87 ±0.34,1.54 ±0.15 at postoperative 12 hours and 0.99 ±0.05,2.02 ±0.27,1.51 ±0.11at postoperative 24 hours,with statistically significant differences among 3 groups (F =22.08,46.71,P ＜ 0.05).(7) Results of ELISA:the level of HA in the non-congestion group,congestion group and congestion + splenectomy group was respectively (149 ± 9) ng/L,(200 ± 19) ng/L,(174 ± 9) ng/L at postoperative 12 hours and (136 ± 16) ng/L,(202 ± 13) ng/L,(91 ± 11) ng/L at postoperative 24 hours,with statistically significant differences among 3 groups (F =19.23,34.68,P＜0.05).Conclusions On the basis of extended liver resection,a wide range of liver congestion through increasing PVP causes hepatic microcirculation disorders,hypoxia,inflammation,vacuoles degeneration cells,increased cells apoptosis,aggravated damage of liver function and increased mortality of rats.Splenectomy could reduce PVP and then improve the liver tissues damage caused by liver congestion,meanwhile,increase the survival rate of rats.

Objective To evaluate the clinical results of free vascularized fibular grafting (FVFG) for the treatment of osteonecrosis of the femoral head (ONFH).Methods From July,2009 to January,2013,85 cases (120 hips) of ONFH were treated with free vascularized fibular grafting.These cases included 61 males (87 hips) and 24 females (33 hips) with an average age of 36.5 years (22-51 years);7 hips (Ⅰ A 2 hips,Ⅰ B 3 hips,Ⅰ C 2 hips) were at stage Ⅰ,98 hips (Ⅱ A 24 hips,ⅡB 39 hips,ⅡC 35 hips) at stage Ⅱ and 15 hips (ⅢA 9 hips,ⅢB 4 hips,Ⅱ C 2 hips) at stage]Ⅲ according to the classification system of Association Research Circulation Osseous (ARCO).The mean preoperative Harris hip score was (60.21±6.85) points (42-71 points),The follow-up items included the X-ray examination,the Harris scores of the hip,and the evaluation of the complications.Results Eighty-three cases (117 hips) were followed up.The average duration of follow-up was 25 months (range from 12 months to 42 months).The mean postoperative Harris hip score was increased to (81.26±5.84) points (67-91 points) by the end of the follow-up,compared with the preoperation,the score improved significantly,the difference was statistically significant (P＜0.05).Comparing with postoperative X-ray,101 hips (86.3％) were improved,12 hips (10.3％) had no significant changes and deterioration occurred in 4 hips (3.4％).Conclusion The free vascularized fibular grafting is an effective method for treating osteonecrosis of and preventing the collapse of the femoral head.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; methods ; DNA, Neoplasm ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Mutation Rate ; Receptor, Epidermal Growth Factor ; genetics

Objective:To investigate the effect and feasibility of an in vitro three-dimensional model for angiogenesis of Kaposiform hemangioendothelioma (KHE). Methods:In April 2019, a 4-month-old boy with KHE received surgery in the department of hemangioma of Henan Provincial People’s Hospital. The tumor tissue was used for the angiogenesis model. Treated tissue was planted in a fibrin gel culture system constructed by a double-layer sandwich method to establish a three-dimensional angiogenesis model. Optimum humidity and constant temperature was provided for the culture system, and the medium was changed every other day. The growth of model was observed every day. When the model became stable, well-grown models were made into frozen sections for HE and immunohistochemistry staining.Results:The model formed newborn vascular buds within 2-4 days and became stable in 12-16 days. The newborn vessels staggered into networks and showed obvious lumen-like structure. Apoptosis started after 26-30 days in most models, while no apoptosis appeared until 54 days in some ones. Morphological observation, HE staining, and immunohistochemistry examination confirmed that KHE endothelia have formed the new vessels.Conclusions:The culture system of fibrin gel is applicable for three-dimensional angiogenesis model of Kaposiform hemangioendothelioma. It is characterized with short culture time, long survival time, simple operation, low cost, and easy intervention.

Objective:To investigate the effect and feasibility of an in vitro three-dimensional model for angiogenesis of Kaposiform hemangioendothelioma (KHE). Methods:In April 2019, a 4-month-old boy with KHE received surgery in the department of hemangioma of Henan Provincial People’s Hospital. The tumor tissue was used for the angiogenesis model. Treated tissue was planted in a fibrin gel culture system constructed by a double-layer sandwich method to establish a three-dimensional angiogenesis model. Optimum humidity and constant temperature was provided for the culture system, and the medium was changed every other day. The growth of model was observed every day. When the model became stable, well-grown models were made into frozen sections for HE and immunohistochemistry staining.Results:The model formed newborn vascular buds within 2-4 days and became stable in 12-16 days. The newborn vessels staggered into networks and showed obvious lumen-like structure. Apoptosis started after 26-30 days in most models, while no apoptosis appeared until 54 days in some ones. Morphological observation, HE staining, and immunohistochemistry examination confirmed that KHE endothelia have formed the new vessels.Conclusions:The culture system of fibrin gel is applicable for three-dimensional angiogenesis model of Kaposiform hemangioendothelioma. It is characterized with short culture time, long survival time, simple operation, low cost, and easy intervention.

Elastic Tissue ; pathology ; Humans ; Lung Neoplasms ; diagnosis ; Pleura ; pathology ; Staining and Labeling

Objective: To investigate the clinicopathologic features and genetic profile of large cell lung carcinoma (LCC) redefined by new classification. Methods: Basing on 2015 WHO classification criteria in redefining large cell lung carcinoma, the expression of specific markers (TTF1, Napsin A, p40, CK5/6, CK, vimentin and ZEB1) was detected by immunohistochemistry and D-PAS staining in 303 surgically-removed lung specimens previously diagnosed as large cell lung carcinoma. The clinicopathologic and genetic characteristics (including EGFR, KRAS, BRAF, ALK and ROS1 gene mutation) were analyzed. Results: Based on the new definition of LCC, 116 cases (116/303, 38.3%) of LCC formerly diagnosed were reclassified as solid adenocarcinoma, 49 cases (49/303, 16.2%) as squamous cell carcinoma, 6 cases (6/303, 2.0%) as adenosquamous carcinoma, 22 cases (22/303, 7.3%) as spindle cell carcinoma and only 110 cases (110/303, 36.3%) as large cell carcinoma. Redefined LCCs were characterized as middle-age (range 40-80), male (102/110, 92.7%) and smoking patients (64/110, 58.2%) with intermediate-advanced stage. Among 110 cases, 9 cases with EGFR mutation and 10 cases with KRAS mutation and 1 case with ALK fusion were found. No BRAF and ROS1 alterations were identified. Conclusions According to the new classification, LCCs formerly diagnosed are mostly reclassified as adenocarcinoma and non-keratinizing squamous cell carcinoma. The newly defined LCC may significantly benefit from clinical therapy.

Objective To establish a rapid detection and identification method for the chemical components of Longshengzhi capsules based on UPLC-Q-TOF/MS.Method UPLC-Q/TOF-MS technology was used to analyze and identify the chemical components of Longshengzhi capsules.The chromatographic column was a Waters Acquity UPLC BEH C18 column(100mm×2.1mm,1.7 μm),and the mobile phase A was 0.1%formic acid water.The mobile phase B was a methanol acetonitrile(1∶1)solution containing 0.1%formic acid(B).The flow rate is 0.3mL/min,the column temperature is 40℃,and the injection volume is 2 μL.The ion source adopts the electric spray ion source.The data is collected in the positive and negative ion modes,and the collection range is m/z 50～1200.The identification and matching are carried out through UNIFI software combined with manual verification of chemical components.Result 87 chemical components were preliminarily and rapidly identified.Conclusion The established method can systematically and rapidly analyze the chemical components in Longsheng leeches,providing a basis for the study of medicinal substances and having important significance for the quality control of Longsheng leeches.