Objective To establish and improve the acute hepatic failure model in pigs induced with D-galactosamine (D-gal),and explore the feasibility of evaluating preclinical artificial liver devices.Methods Nineteen Duroc breeding pigs were divided into 4 groups.Fifteen unanesthetic Duroc breeding pigs out of 19 (5 of each group) received intravenously administration of D-gal at a dose of 1.0,1.25 and 1.5 g/kg body weight,respectively.The remaining 4 pigs which received the same volume of 5% dextrose in water served as controls. Clinical data and survival time of pigs were recorded.Blood samples were collected for dynamic testing of plasma ammonia,prothrombin time,liver and renal functions,blood glucose and L-lactate;liver tissues were sampled for pathological examination.The differences between groups were compared using t test and F test.The survival time of pigs was compared by Kaplan-Meier survival analysis and Log Rank test.Results Twelve hours after administration of D-gal,all pigs presented as acute hepatic failure characterized by progressive increases of levels of plasma ammonia,aspartate aminotransferase (AST),total bilirubin (TBil) and L-lactate,the level of blood glucose marked decreased and prothrombin time prolonged (F= 32.33,F=27.817,F=50.097,F=88.382,F=8.211,F=21.227;all P＜0.01);especially in the pigs which received D-gal at a dose of 1.5 g/kg.Except 2 pigs survived for 168 h,the other 3 pigs which received D-gal at a dose of 1.0 g/kg died within 68-84 h,while all pigs which received D-gal at a dose of 1.25 and 1.5 g/kg died within 33-89 h and 23-47 h,respectively.All pigs presented coma before death and liver histopathological examination indicated massive hepatic necrosis with severe hemorrhage.Conclusions D-gal induced acute hepatic failure model in unanesthetic Duroc breeding pig appears potential reversibility and high reproducibility,which has proper therapeutic window.Thus,this model could be applied to evaluate the therapeutic effects and safety of artificial liver devices.

Objective To investigate the diagnosis and treatment of primary hyperparathyroidism ( PHPT) initially with urinary calculus .Methods The clinical data of 26 patients who diagnosed as PHPT ini-tially with urinary calculus were retrospectively reviewed .Results There were 22 cases with bilateral urinary calculus and 4 cases with unilateral relapsed urinary calculus .Ultrasonography , CT and radionuclide were helpful to determine the location of the neoplasia .24 cases underwent percutaneous nephroscope or ureteroscopy pneu-matic ballistic lithotripsy , 2 cases discharged ureteral calculi by themselves .All patients were performed surgical treatment of PHPT , which was confirmed by intraoperative frozen pathology and postoperative pathological exami -nation.There was great improvement of clinical symptoms after surgical procedures .The preoperative serum calci-um, urine calcium and parathyroid hormone elevated , while serum phosphate decreased .The postoperative indi-cators were just the opposite .The difference had statistical significance ( P<0.05 ) .Conclusions Laboratory investigations and imaging studies are very important to diagnose PHPT initially with urinary calculus .The para-thyroid surgery can remarkably reduce the calculus recurrence and improve renal function .

Objective To explore the characteristic of intelligence, personality and mentality of patients with intractable epilepsy and analyzethe related factors on intelligence. Methods 68 patients with intractable epilepsy and 60 healthy persons were assessed with Raven'sstandard progressive matrices (SPM), Eysenck personality questionnaire (EPQ) and 90 symptoms checklist (SCL-90). The related factors onintelligence were analyzed. Results Compared with controls, the total score and IQ of SPM of the patients decreased significantly (P<0.05).There were 51.4% of intellectual deficiency in patients with intractable epilepsy, which was higher than that of controls (1.7%) (P<0.05).There was no significant difference in score of EPQ and the proportion of personality type between 2 groups (P>0.05). The total score ofSCL-90 and the factor score, such as somatization, interpersonal sensitivity, depression, anxiety and hostility in patients were higher thanthose in controls(P<0.05). Various factors, including the onset age, course of disease, seizure types, frequency, duration, number of drug, thescores of SCL-90 were correlated with the intelligence of epileptic patients (P<0.05). Conclusion The intelligence of many patients with intractableepilepsy were damaged, and many patients were with various psychological barriers. The onset age, course of disease, seizuretypes, frequency, duration and number of drug were important factors related with intellectual disorders in patients with intractable epilepsy.

Objective To establish normally immortalized porcine hepatocyte lines by ectopic expression of simian virus 40 large T (SV40LT) antigen and the human telomerase reverse transcriptase(hTERT). Methods Primary porcine Hepatoeyte cells were transfeeted with recombinant retrovirus containing SV40LT or hTERT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Immortalized porcine hepatocyte was confirmed by examination. Results The morphological phenotype of the transfected cells was similar to the primary porcine hepatocyte. One clone, HepLP, has been maintained in cultue for half year, and expanded by more than 60 passages. SV40 LT and hTERT could be detected in transfected porcine hepatocyte. Pig albumin mRNA was also detected by RT-PCR. No tumor formation occurred when HepLP cells were injected into Balb/c nude mice. Conclusions The immortalized, nontumorigenic, porcine hepatoeytes maintained the properties of porcine primary hepatocytes such as the albumin secretion. This generation of immortalized porcine hepatocyte may be helpful for bioartifical liver support system, hepatocytes transplantation, drug/toxicological studies, and liver biologic studies.

AIM:Early calcification of atherosclerotic plaques are colocalized with macrophage and high mobility group box 1 (HMGB1), a cytokine associated with biomineralizing process under physiological and pathological conditions .Our study aims to evaluate whether HMGB1 induces ectopic mineralization via promoting the secretion of matrix vesicles ( MVs) from macrophages .METHODS:HMGB1 was added to the medium of macrophages , the secretion of MVs in the supernatant was tested by flow cytometry analysis .The mineral deposition in calcifying medium was detected by Alizarin Red staining and von Kossa staining .Transmission electron microscopy showed the formation of hydroxyapatite crystals in MVs .Then we subcutaneous injection into mice with MVs to induce regional minera-lization.RESULTS:HMGB1 significantly promoted secretion of MVs from macrophages as raveled by flow cytometry analysis .TNAP activity, considered as a marker of MVs maturation , was higher in HMGB1-induced MVs compared to the control-MVs.HMGB1-MVs also led to mineral deposition in an in vitro MVs-collagen mineralization model .Subcutaneous injection into mice with MVs derived from HMGB1-treated cells showed a greater potential to initiate regional mineralization .Mechanistic experiments revealed that HMGB 1 activated neutral sphingomyelinase 2 ( nSMase2 ) that involved the receptor for advanced glycation end products ( RAGE ) and p38 MAPK (upstream of nSMase2).Inhibition of nSMase2 with GW4869 or p38 MAPK with SB-239063 prevented MVs secretion and min-eral deposition .CONCLUSIONS: HMGB1 induces MVs secretion from macrophages at least in part , via the RAGE/p38 MAPK/nSMase2 signaling pathway .Our findings thus reveal a novel mechanism by which HMGB 1 may participated in the early calcification of atherosclerotic plaques .

Objective To establish immortalized human hepatocyte lines for studies of bioartificial liver,hepatocyte transplantation,and drug metabolism in vitro.Methods Primary human hepatocytes were isolated by 4-step perfusion technique with collagenase and transfected with recombinant retrovirus containing Simian virus 40 large T antigen(SV40 LT).Subsequently,immortalized human hepatocytes were evaluated by analysis of gene expression and functional characteristics in vitro.Results Two immortalized human hepatocyte lines,HepLi2 and HepLi3,were obtained after primary human hepatocytes being infected by SV40 LT containing recombinant retrovirus for 3-4 weeks.The immortalized human hepatocytes showed classical appearance of hepatocyte observed by phase contrast microscope.The protein expression of SV40 LT in HepLi2 and HepLi3 cells were detected by Western blotting.The mRNA expressions of albumin(Alb),glutathione S-transferase(GST-p),human blood coagulation factor X(HBCF-X)and β-actin in HepLi2 and HepLi3 cells were detected by reverse transcription polymerase chain reaction(RT-PCR),and the mRNA expressions of cytochrome(CY)450 subtypes(CYP3A5,CYP2E1,CYP2C8-19 and CYP3A4)in HepLi2 and HepLi3 cells were also observed by RT-PCR.Levels of alanine transaminase (ALT),lactate dehydrogenase(LDH)and Alb were detected in the supernatant of immortalized human hepatoeyte culture.Conclusions The immortalized human hepatocyte lines have the biological characteristics of primary human hepatocytes and have the CYP450 functions of hepatocytes,which may be heIDful for the studies of bioartificial liver,heoatocvte transplantation and drug metabolism in vitro.

Objective:To investigate the clinical characteristics, risk stratification, thrombolytic effects and prognosis of 110 patients with acute pulmonary embolism (PE) treated with thrombolysis.Methods:The clinical data of 110 patients with PE admitted to Beijing University People's Hospital from May 2009 to March 2019 were retrospective analyzed. The clinical data including general information, symptoms and signs, blood pressure, artery blood gas, coaglulation, and radiography were collected. Inclusion criteria: high-risk and intermediate high-risk group. Exclusion criteria: intermediate low-risk and low-risk group. According to the prognosis and risk stratification, the patients were divided into survival group and non-survival group, high-risk group and intermediate high-risk group. The indicators above were compared between with χ 2 test, t test or nonparametric test where appropriate. Results:Of the 110 patients with PE, 49 patients were male and 61 female with an average age of 65±16 years old; and 12 patients were in the high-risk group and 98 in the intermediate high-risk group. The respiratory rate of the high-risk group was higher, and blood pressure, PO 2, SaO 2 before thrombolysis were more lower than the intermediate high-risk group ( P<0.05). One hundred and nine patients were treated with systemic recombinant tissue plasminogen activator (rtPA), 70 patients with 50 mg, and 39 patients with 100 mg. One patient, who was contraindicated to systemic thrombolysis (with active vagina bleeding), was treated with interventional local thrombolysis; another 5 patients treated with interventional local thrombolysis because the clinical symptom were not improved markedly. One hundred and two patients survived and 8 patients died, among which, 3 patients were in the high-risk group and 5 in the intermediate high-risk group. The age, heart rate, respiration rate of the non-survival group were higher than those in the survival group, and the PO 2 before thrombolysis, PCO 2 after thrombolysis were lower ( P<0.05). Bleeding complication were occurred in 22 patients: 18 patients with minor bleeding, such as bleeding gums, skin ecchymosis, and 4 patients with moderate-severe bleeding, such as cerebral hemorrhage, abdominal bleeding, gastrointestinal bleeding, and vagina bleeding. Thirteen of 70 patients in the 50 mg group and 9 of 39 patients in the 100 mg group occurred bleeding complication. The bleeding complication of the low dose group was lower than that of the standard dose group ( P<0.05). Conclusions:Thrombolysis is first-line therapy to high-risk PE. Thrombolysis is safe and effective in the intermediate high-risk group with a lower incidence rate of bleeding complication.

Objective To investigate the effectiveness of T1 mapping on gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA-enhanced) MRI for the assessment of liver function.Methods One hundred and twenty six patients with clinically suspected focal liver lesions and chronic viral hepatitis B underwent MRI were prospectively enrolled.Patients were divided into four subgroups as follows: chronic viral hepatitis B (n=22), liver cirrhosis with Child-Pugh A (n=52), Child-Pugh B(n=41),Child-Pugh C(n=11).Twenty three healthy volunteers with normal liver function were enrolled as control group.Non-enhanced and Gd-EOB-DTPA enhanced MRI of liver were performed in all subjects.Look-Locker sequences with exactly the same scan parameters and geometry position(the level of porta hepatis) were performed pre and post-contrast separately at 5, 10, 15 and 20 minutes after Gd-EOB-DTPA administration.T1 relaxation times and reduction rates of T1 relaxation times[ΔT1(%)]of the liver parenchyma were measured and calculated.One-way ANOVA was used to compare T1 relaxation times and ΔT1(%) for control group, chronic viral hepatitis B group, liver cirrhosis with Child-Pugh A group, Child-Pugh B group,and Child-Pugh C group.ROC curve analysis was performed to compare the diagnostic performance of T1 relaxation times and ΔT1(%) values in discriminating control group + chronic viral hepatitis B group + liver cirrhosis with Child-Pugh A group from Child-Pugh B + C group. Results T1 relaxation times and ΔT1(%)showed significant difference(P<0.05)among control group and different liver function groups. T1 relaxation times and ΔT1(%) of both liver cirrhosis with Child-Pugh B group and Child-Pugh C group were significantly different(P<0.05)in comparison with those of control group,chronic viral hepatitis B group and liver cirrhosis with Child-Pugh A group at all time points.T1 relaxation times of the control group,chronic viral hepatitis B group,liver cirrhosis with Child-Pugh A group and Child-Pugh B group reduced with the scanning time increase,ΔT1(%)raised with the scanning time increase.T1 relaxation times progressively increased from control group to Child-Pugh C group at every time point.ΔT1(%)showed a constant decrease from control group to Child-Pugh C group at all time points.The areas under ROC curve of T1 relaxation time pre and post-contrast at 5,10,15 and 20 minutes for assessment of liver function were 0.817,0.952,0.950,0.946,and 0.949 respectively.The areas under ROC curve of ΔT1(%)post-contrast at 5, 10, 15 and 20 minutes for evaluation of liver function were 0.873, 0.876, 0.885, and 0.898, respectively. Conclusion Gd-EOB-DTPA-enhanced T1 mapping MRI is useful for the evaluation of liver function, and helpful for distinguishing patients with moderate and severe liver damage from normal and mild liver damage.

Objective To evaluate the value of T1 mapping in Gd-EOB-DTPA-enhanced MRI for the assessment of liver function with HBV-related cirrhosis according to the model for end-stage liver disease (MELD) score.Methods 158 patients with HBV-related cirrhosis were included in this prospective study,and divided into MELD score ≤10 (n =103) group and MELD score ＞ 10 (n =55) group.All patients un derwent non-enhanced and Gd-EOB-DTPA enhanced MRI of liver,and T1 mapping was performed using Look-Locker sequences with the same scan parameters and geometry position (the level of porta hepatis) preand post-contrast at 5,10,15 and 20 minutes after Gd-EOB-DTPA administration.T1 relaxation times of the liver were measured and reduction rates of T1 relaxation times (△T1) were calculated.Independent samples t test was performed to compare T1 relaxation times and △T1 between MELD score≤ 10 and MELD score ＞ 10 groups.Receiver operating characteristic curve (ROC) analysis were done to differentiate the diagnostic performance of T1 relaxation times and △T1 between MELD score ≤ 10 and MELD score ＞ 10 groups.Pearson correlation analysis was used to analyse the correction between T1 relaxation times,△T1 and MELD scores.Results T1 relaxation times pre-and post-contrast at 5,10,15 and 20 minutes and △T1 post-contrast at 5,10,15 and 20 minutes of MELD score≤10 group were (889.3 ±91.2) ms,(377.5 ± 55.0) ms,(350.8±61.2)ms,(328.0±69.4)ms,(302.7±73.7)ms,(57.4±5.6)％,(60.4± 6.5) ％,(63.0 ± 7.3) ％ and (65.9 ± 7.8) ％,respectively,and those of MELD score ＞ 10 group were (936.6 ±95.4) ms,(460.2 ±68.5) ms,(457.5 ±94.5) ms,(453.4 ± 116.4) ms,(444.6 ± 134.6) ms,(50.8 ± 5.7) ％,(51.3 ± 7.9) ％,(51.8 ± 10.3) ％ and (52.8 ± 12.2) ％,respectively,and T1 relaxation times and △T1 at all time points were significantly different (P ＜ 0.05) between the two groups.The areas under ROC curve of T1 relaxation time pre-and post-contrast at 5,10,15,20 minutes and △T1 post-contrast at 5,10,15,20 minutes for differentiating MELD score ≤ 10 and MELD score ＞ 10 groups were 0.638,0.824,0.832,0.832,0.830 and 0.795,0.814,0.820,0.825,respectively.The correlation coefficients between T1 relaxation time pre-and post-contrast at 5,10,15,20 minutes,△T1 post-contrast at 5,10,15,20 minutes and MELD scores were 0.256,0.499,0.540,0.538,0.548,-0.412,-0.495,-0.507 and-0.527,respectively.Conclusions T1 mapping on Gd-EOB-DTPA-enhanced MRI is helpful for evaluating liver function with HBV-related cirrhosis.T1 relaxation times post-contrast on different time points were equally accurate as △T1.T1 relaxation times post-contrast and △T1 were superior to T1 relaxation times pre-contrast.

Tumor-induced osteomalacia (TIO), also known as oncogenic osteomalacia, is a rare paraneoplastic syndrome characterized by hypophosphatemia resulting from decreased tubular phosphate reabsorption, with a low or inappropriately normal level of active vitamin D. The culprit tumors of TIO could produce fibroblast growth factor 23 which plays a role in regulating renal Pi handling and 25-hydroxyvitamin D 1α-hydroxylase activity. Chronic hypophosphatemia could eventually lead to inadequate bone mineralization, presenting as osteomalacia. The diagnosis should be considered when patients manifest as hypophosphatemia and osteomalacia, or rickets and needs to be differentiated from other disorders of phosphate metabolism, such as the inhereditary diseases like X-linked hypophosphataemic rickets, autosomal dominant hypophosphataemic rickets, autosomal recessive hypophosphataemic rickets and acquired diseases like vitamin D deficiency. Localization of responsible tumors could be rather difficult since the vast majority are very small and could be everywhere in the body. A combination of thorough physical examination, laboratory tests and imaging techniques should be applied and sometimes a venous sampling may come into handy. The technology of somatostatin-receptor functional scintigraphy markedly facilitates the localization of TIO tumor. Patients undergoing complete removal of the causative neoplasm generally have favorable prognoses while a few have been reported to suffer from recurrence and metastasis. For those undetectable or unresectable cases, phosphate supplements and active vitamin D should be administrated and curative intended radiotherapy or ablation is optional.
Calcification, Physiologic ; Diagnosis ; Fibroblast Growth Factors ; Humans ; Hypophosphatemia ; Metabolism ; Neoplasm Metastasis ; Osteomalacia ; Paraneoplastic Syndromes ; Physical Examination ; Prognosis ; Radionuclide Imaging ; Radiotherapy ; Recurrence ; Rickets ; Vitamin D ; Vitamin D Deficiency