Objective To construct a novel bioartificial liver (BAL) system and evaluate its functions in vitro. Methods Chinese experimental minipig hepatocytes were isolated by in situ recirculating collagenase perfusion method and 1.0?10~(10) hepatocytes were cultured in serum-free medium with restriction of(attachment), and using spinner method to form hepatocyte(spheroids).cted by inoculating the hepatocyte(spheroids) into cell circuit of a hollow fiber bioreactor from BIOLIV A3A. Observing the number and viability of the(hepatocytes), the changes of alanine aminotransferase (ALT), total bilirubin (TBI), albumin (ALB) in(circulating) hepatocyte suspension and(RPMI1640) medium; in addition, lidocaine metabolism test was(determined),(during) 6h circulation of the system. (Results) There were no significant differences in number and viability of the hepatocytes before and after 6h(circulation). The BAL system has relatively strong albumin synthesis and lidocaine(metabolism) functions. (Conclusions) The BAL system that we developed had ability to support liver functions and could be used in the treatment of liver failure, or to provide temporary liver support for candidates of liver(transplantation).

Objective:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

Objective To optimize the extraction process for the polysaccharide from fermented Cordyceps Sinensis powder with response surface methodology.Methods We selected the factors and levels on the basis of single factor experiment,and then designed the experiment with 3 factors and 3 levels based on the principle of Box-Behnken's design.Results The effect of the factors of extraction temperature,time and solid-liquid ratio on extraction ratio was in decreasing sequence.The optimal extraction technology obtained through the classical analysis was as follows:extraction temperature at 95 ℃,the ratio of solid to liquid being 1 ∶ 21,and extraction for 73 min.Under this condition the theoretical extraction rate was 4.31％ and the actual extraction rate was (4.20 ± 0.1)％.Conclusion The obtained values agree with the predicted values of the mathematic models,and the Box-Behnken experimental design is suitable for optimizing the extraction of the polysaccharide from fermented Cordyceps Sinensis powder.

Objective Using polysulfon fibers, a new bioartificial liver was developed. This study was to evaluate the efficacy of this bioartificial liver in the support of a disfunctioned liver. Methods Hepatocytes were procured from swine using Seglen′s methods. The bioartificial liver was constructed based on polysulfon bioreactor with a procurement of 10 10 hepatocytes, and was applied in 12 acute liver failure patients for 14 sessions. Each BAL treatment lasted 6 hours. The general conditions of the patients and the biochemical parameters were evaluated. Results After treatment with bioartificial liver, ammonia, prothrombin time and total bilirubin level significantly decreased (all P

Objective To establish the condition cultrue cell system and co-culture cell system with SKOV3/PM4,HUVEC and to study the changes of their biological characteristics. Methods The cells of SKOV3/PM4 and HUVEC were labeled with green and red fluorescent respectively. The cell supernatant of SKOV3/PM4 and HUVEC were collected respectively as the condition medium(e.g:the cell supernatant of HUVEC cells was used as SKOV3/PM4 condition medium)and to establish the condition cultrue cell system and the co-culture cell system of the two cell lines. In the condition cultrue cell system, The morphological changes of cells were observed by HE staining to calculate the mitotic index. The ultrastructural changes of the two cells were observed by transmission electron microscopy(TEM). The growth curve of the cells was determined by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry was used to analyzed the cell cycles.In the co-culture cell system, the interaction of the two cells were detected by laser scanning confocal microscope(LSCM). The expression of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9 (MMP-9) were detected by gelatin zymography. Results Compared with the single culture SKOV3/PM4, the cells which cultured in HUVEC condition medium showed the increase of pseudopodia and nuclear division,the mitotic index respectively were [(4.8 ± 0.8)%,(11.2 ± 0.3)%;P<0.05]. The growth rate was significantly increased. In cell cycles, it showed the declined cell ratio of G0/G1 phase, respectively[(69.4±3.6)%, (48.4±4.6)%;P<0.05] and the raised cell ratio of G2/M phase, respectively [(5.2±1.6)%, (24.9±2.2)%;P<0.05]. Compared with the single culture HUVEC,the cells which cultured in SKOV3/PM4 condition medium showed the significant morphological change and vacuolization in the cytoplasm, Nuclear division was increased and the mitotic index respectively were [(2.7±0.5)%, (5.7±0.6)%;P<0.05]. The growth rate was slightly declined. In cell cycles, it showed the raised cell ratio in G0/G1 phase, respectively [(51.4 ± 2.2)%,(79.0 ± 4.1)%;P<0.05] and the declined cell ratio in G2/M phase, respectively [(19.1±1.2)%, (3.3±0.5)%;P<0.05]. After co-culture for 48 hours, spontaneous fusion between SKOV3/PM4 and HUVEC cell was observed by the laser confocal microscope. Gelatin zymography assay showed that MMP-2 was not expressed in HUVEC cells, low-expressed in SKOV3/PM4 cells and high-expressed in the co-culture SKOV3/PM4+HUVEC cells. The expression of MMP-2 in co-culture SKOV3/PM4+HUVEC cells and SKOV3/PM4 cells respectively were 1 885 ± 84 and 1 209 ± 114 (P<0.05). But there were no MMP-9 expression in HUVEC cells, SKOV3/PM4 cells, and the co-culture SKOV3/PM4+HUVEC. Conclusion The characteristics of SKOV3/PM4 and HUVEC show significant changes after condition culture and co-culture, it may involve in the microenvironment of the cells and the intercellular crosstalk pathway.

Objective To compare the effect of pericardial devascularization with that of the modified Sugiura procedure in management of portal hypertension. Methods From 1990 to 2008, 236patients with portal hypertension underwent operations including pericardial devascularization in 147and modified Sugiura in 89 in our hospital. Results There were 12 perioperative deaths (8.2 % ), and 2 rebleedings (2 % ) in the pericardial devascularization group, and 7 perioperative deaths (7.9 % ) and 2 rebleedings(3.4 % ) in the modified Sugiura group. The follow-up rate was 91.9 % in the pericardial devascularization group and 87.8% in the modified Sugiura group respectively, in a period from 6 months to 19 years. The 1-, 3-and 5-year rebleeding rates were 5.7%,15.2% and 25.5% in the pericardial devascularization group and 6.9%, 16.3%, 29.5 % in the modified Sugiura group, respectively. The 1-, 3- and 5-year survival rates were 87.8% ,79.1% and 69.7% in the pericardial devascularization group and 95.8 %,85.0%, 76.9 % in the modified Sugiura group, respectively. Conclusion Modified Sugiura procedure and pericardial devascularization have differences in perioperative mortality as well as rebleeding and survival rates.

OBJECTIVE:To establish a method for the content determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules. METHODS:HPLC method was adopted. The determination was performed on WondasilTM-C18 column with mobile phase consisted of acetonitrile-0.4% phosphoric acid solution(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 30 ℃,and the detection wavelength was set at 254 nm. The sample size was 5 μL. RESULTS:The linear range of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid were 0.014-0.560 μg (r＝0.999 2),0.022-0.880 μg(r＝0.999 6),0.010-0.400 μg(r＝0.999 9),0.016-0.640 μg(r＝0.999 9),0.011-0.440 μg(r＝0.999 1), respectively. The limits of quantitation were 0.752,0.647,0.483,0.372,0.242 mg/L;the limits of detection were 0.253,0.185, 0.157,0.162,0.098 mg/L. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The average recoveries were 97.4%-100.9%(RSD＝1.4%,n＝6),95.7%-99.0%(RSD＝1.1%,n＝6),95.1%-98.1%(RSD＝1.3%,n＝6),96.4%-99.8%(RSD＝1.4%,n＝6),97.8%-100.4%(RSD＝1.0%,n＝6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules.

Objective:To investigate the level of psychological stress, anxiety and posttraumatic stress disorder (PTSD) of military medical students under the condition of normalized prevention and control of the epidemic.Methods:A total of 225 students from a military university were selected by random sampling method. Chinese perceived stress scale (CPSS), Perceived social support scale (PSSS), Self-rating anxiety scale (SAS) and PTSD Checklist-5 (PCL-5) were used to measure the mental health of the subjects and influencing factors. SPSS 26.0 was used to conduct independent sample t test and ANOVA analysis. Pearson correlation analysis was conducted, and multiple regression equation was used to further analyze the relationship between psychometric factors. Results:The SAS scores of 22.2% (50/225) of the subjects and PCL-5 scores of 30.7% (69/225) of the subjects reached the positive standard. The results of correlation analysis showed that the total score of PSSS was negatively correlated with CPSS ( r=-0.315, P<0.01), SAS ( r=-0.336, P<0.01) and PCL-5 ( r=-0.137, P<0.05) scores. The score of family support negatively predicted the level of psychological stress ( B=-0.595, P<0.001), anxiety ( B=-0.635, P<0.001) and PTSD level ( B=-0.769, P=0.006) of the students. Conclusion:Social support, especially family support, has a significant protective effect on psychological stress, anxiety and PTSD during the period of normalized prevention and control of the epidemic.

Objective:To discuss the ultrasonographic features of ganglioneuroma(GN), ganglioneuroblastoma-intermixed(GNBi), ganglioneuroblastoma-nodular(GNBn), neuroblastoma(NB) in children and to improve the ultrasound diagnostic accuracy of four kinds of neuroblastic tumors.Methods:Two hundred and seventy-nine patients with neuroblastic tumors(23GN, 44GNBi, 86GNBn, 126NB) confirmed by surgery and pathological diagnosis in Children′s Hospital Affiliated to Capital Institute of Pediatrics from June 2014 to February 2019 were retrospectively analysed. The ultrasonographic data were summarized and compared with pathological results.Results:The median age were 6.9 years in GN group, 3.9 years in GNBi group, 3.0 years in GNBn group and 2.0 years in NB group. The median values of serum neuron-specific enolase(NSE) were 6.3 μg/L in GN group, 9.6 μg/L in GNBi group, 22.6 μg/L in GNBn group and 40.7 μg/L in NB group respectively. There was no distant metastasis of GN, while 9% GNBi, 26% GNBn and 36% NB had distant metastasis; GN and GNBi group had no invasion to adjacent tissues and organs, while 9% GNBn and 15% NB lesions had invasion to adjacent tissues and organs; 91% GN, 70% GNBi, 58% GNBn lesions had complete capsule, while 44% NB lesions had incomplete capsule and 28% NB had no capsule; 96%GN, 57%GNBi lesions were round or oval, while 57%GNBn, 60%NB lesions showed irregular shape; 96%GN showed homogeneous hypoechoic, 75% GNBi had "cloudy" inhomogeneous echoe, while hyperechoic nodules were seen in 59% GNBn and 75% NB lesions; 65%GN lesions contained discrete and punctate calcifications, while 27%GNBi, 29%GNBn, 25%NB lesions contained coarse calcifications; 100%GN, 91%GNBi, 91%GNBn lesions had little to moderate blood flow, while 33%NB lesions had abundance blood flow; 4%GN, 41%GNBi, 51%GNBn, 49%NB lesions surrounded blood vessels; 13%GN, 5%GNBi, 6%GNBn, 8%NB lesions had spinal canal invasion; GN had no lymph node metastasis, but 48%GNBi, 59%GNBn, 56%NB lesions had lymph node metastasis.Conclusions:The ultrasonographic characteristics of GN are largely different from GNBi, GNBn and NB; There are some differences in ultrasonographic features of GNBi, GNBn and NB, but some of them are difficult to identify.

Objective:To explore the surgical procedure and clinical effect of the free Flow-through superficial peroneal artery flap for repairing the digit injury with defect of proper palmar digital artery.Methods:From June, 2015 to December, 2019, free Flow-through superficial peroneal artery flap was used to repair the digit injury and to bridge the proper digital artery in 7 digits of 7 patients. There were 2 thumbs, 3 index fingers and 2 middle fingers. The size of defects on digits ranged from 2.5 cm×3.0 cm to 5.0 cm×7.0 cm. The defects of digit proper artery were 1.0 to 3.0 cm. The size of flaps was from 3.0 cm×3.5 cm to 5.5 cm×8.0 cm. The donor areas of the flap were directly sutured or covered with skin graft. Postoperative supportive treatments were given, such as infection prevention, antispasmodic, anticoagulant, analgesia and fluid infusion. The patients were kept in bed for 1 week after surgery. Monthly follow-up review were conducted and the patients were kept with the rehabilitation exercises under medical guidance.Results:All the patients entered postoperative followed-up for 6 to 18 months, with an average of 8 months. All flaps survived without any adverse event. All wounds achieved stage-one-healing. The flaps appeared in good colour, texture, elasticity and the plumps of the digit without obvious bloating. There was no obvious swelling and atrophy of the digits. The skin temperature was normal. According to the Standard for Evaluation of Upper Limb Function by the Hand Surgery Society of Chinese Medical Association, 3 digits were excellent and 4 were good. There was no obvious scar at the donor site of shank. The donor site had a good appearance and the limb function was not affected.Conclusion:The free Flow-through superficial peroneal artery flap is an ideal material to repair the defect of digit with the defect of proper digital artery. It has the advantages of simple surgical procedure, reliable blood supply and satisfactory appearance. The defect of proper digit artery can be repaired at the same time of the surgical procedure. The blood supply, appearance and function of the digits could be well recovered and the donor site is not affected.