Advanced hepatocellular carcinoma has a high degree of malignancy and poor prognosis. Studies showed that there is a close relationship between the progression of hepatocellular carcinoma and the immune status in tumor microenvironment. Adoptive cell therapy showed anti-tumor effects and improve immunosuppression by infusing patients with activated specific immune cells, which become a central issue in tumor therapy and shown promising effects in the treatment of various malignant tumors, indicating great application potential. Adoptive cell therapy based on neoantigen may become a new hot spot in the treatment of hepatocellular carcinoma, and their application, safety and effectiveness evaluation, efficacy prediction and assessment have become urgent issues to be solved. The purpose of this article is to introduce the progress related to adoptive cell therapy for advanced hepatocellular carcinoma and elaborate the problems that need to be solved in the future.

Objective:To analyze the anti effect of chimeric antigen receptor (CAR)-T cells targeting hepatitis B surface antigen (HBsAg) on hepatocellular carcinoma cells.Methods:HBsAg-CAR gene was transduced into T cells (obtained from the blood of healthy donors) through a lentiviral vector. CD19-CAR-T cells were included as mock group, and untransduced T cells were included as control group. Cells of the three groups were co-cultured with hepatocellular carcinoma cells expressing HBsAg or not to detect the anti effect and releasing level of anti-tumor cytokines (tumor necrosis factor-α, interferon-γ, interleukin-2). Subcutaneous xenograft PLC/PRF/5 tumor model using NPG mice were established and HBsAg-CAR-T cells (experimental group, n=5) or untransfected T cells (control group, n=5) were injected through tail vein. Tumor volume was measured 15 days after injection. Results:HBsAg-CAR-T cells proliferation was good under in vitro culture, and the expression rate of CAR was stable. After co-cultured with hepatocellular carcinoma cells expressing HBsAg, the level of anti-tumor cytokines released by HBsAg-CAR-T cells was significantly higher than that of the other two groups of T cells, and the difference was statistically significant (all P<0.05); the anti rate of HBsAg-CAR-T cell group on HBsAg-positive hepatocellular carcinoma cells was significantly higher than that of the other two groups, and the difference was statistically significant (all P<0.05). The tumor volume of NPG mice in the experimental group was (250.8±62.8) mm 3, which was lower than that of the control group (757.5±102.6) mm 3, and the difference was statistically significant ( P<0.05). Conclusion:HBsAg-CAR-T cells can specifically recognize and kill HBsAg-positive hepatocellular carcinoma cells and release high level of anti-tumor cytokines.

Objective To evaluate the effects of the serum of patients with obstructive jaundice on myogenic differentiation of human pulmonary microvascular endothelial cells (PMVECs). Methods Hu?man PMVECs were isolated and then subcultured. The cultured PMVECs were incubated with the serum of patients with obstructive jaundice or with the serum of healthy volunteers. At 24, 48 and 72 h of incubation (T1?3), the inverted microscope was used to observe the morphology of primary PMVECs. The expression of muscular proteins ( alpha?smooth muscle actin [α?SMA ] , smooth muscle?mysion heavy chain [ SM?MHC] , capolnin) in PMVECs was detected using Western blot analysis. Results The expression of cal?ponin andα?SMA was negative, and a few SM?MHC proteins were expressed when PMVECs were incubated with the serum of healthy volunteers; the expression of calponin, α?SMA and SM?MHC was positive when PMVECs were incubated with the serum of patients with obstructive jaundice. Compared with the serum of healthy volunteers, the expression of SM?MHC was significantly up?regulated when PMVECs were incubated with the serum of patients with obstructive jaundice (P<0.05). The expression of calponin, α?SMA and SM?MHC was significantly up?regulated at T2,3 compared with that at T1 , and at T3 compared with that at T2 when PMVECs were incubated with the serum of patients with obstructive jaundice ( P<0.05) . Conclusion The serum of patients with obstructive jaundice promotes myogenic differentiation of human PMVECs, which is probably one of the mechanisms underlying intrapulmonary microvascular dilatation.

Objectives We sought to determine the factors that predicted in-hospital heart failure(HF)in patients undergoing successful primary percutaneous coronary intervention(PCI)for ST-segment elevation myocardial infarction(STEMI). Methods The clinical and angiographic data were retrospectively reviewed in patients undergoing successful primary PCI for their ifrst STEMI. According to the occurrence of in-hospital HF, patients were divided into HF group and non-HF group. The incidence and predictors of in-hospital HF and its impact on prognosis were determined. Results A total of 834 patients were included, among them 94 patients (11.3%) were in the HF group and 740 patients(88.7%) were in the non-HF group. The mean age was (62.9±12.9) years and 662 patients (79.4%) were male. All-cause mortality at 30 days was signiifcantly higher in the HF group than in the non-HF group (24.5%vs. 1.5%, P<0.001). In Cox regression analysis, left anterior descending artery (LAD) as the culprit vessel (HR 2.173, 95% CI 1.12~4.212, P=0.022), ln 24 h NT-proBNP (HR 1.904, 95%CI 1.479~2.452, P<0.001), 24 h hsCRP≥11.0 mg/L (median) (HR 2.901, 95%CI 1.309~6.430, P=0.009) and baseline serum glucose (HR 1.022, 95%CI 1.000 ~ 1.044, P=0.046) were independent predictors of in-hospital HF. Receiver operator characteristic analysis identiifed 24 h NT-proBNP ≥ 1171 pg/ml (c=0.883, P < 0.001) and 24 h hsCRP ≥ 13.5 mg/L (c=0.829, P < 0.001) were the best cut-off values in discriminating in-hospital HF with a sensitivity and speciifcity of 92.5%and 76.8%for 24 h NT-proBNP, 86.0%and 77.0%for 24 h hsCRP, respectively. Even among patients with LAD as the culprit vessel, the incidence of in-hospital HF was only 0.4%in patients whose 24 h NT-proBNP was<1171 pg/ml and 24 h hsCRP was<13.5 mg/L;while the incidence of in-hospital HF was up to 60.9%in patients whose 24 h NT-proBNP≥1171 pg/ml and 24 h hsCRP≥13.5 mg/L (P<0.001). Conclusions The incidence of in-hospital HF was still high in STEMI patients even after successful primary PCI. Patients with in-hospital HF had poor prognosis. LAD as the culprit vessel, hsCRP, NT-proBNP and baseline serum glucose were independent predictors of in-hospital HF. Assessment and combined use of different serum biomarkers were effective methods to estimate the risk of in-hospital HF in STEMI patients undergoing primary PCI.

Objective:To establish an HPLC method for the determination of thymol in thymol alcoholic solutions. Methods: An Agilent Eclipse XDB-C18(250 mm ×4.6 mm,5μm) column was used with the mobile phase of methanol-water (65∶35), the flow rate was 1. 0 ml·min-1 . the detection wavelength was 275nm, the injection volume was 10μl, and the column tenperature was 25℃. Re-sults:A good linear correlation of thymol was observed within the range of 60-160 μg·ml-1(r=0. 999 5). The average recovery was 101. 59% with RSD of 1. 39%(n=9). Conclusion:The method is quick, simple and accurate, which can be used in the determina-tion of thymol alcoholic solutions with good selectivity and sensitivity.

We describe a case of bilateral thyroid carcinoma in a 48-year-old woman. She was admitted to our hospital as a case of cervical nodular goiter. Multifocality of the thyroid nodules were evaluated by preoperative ultrasonography. The patient received bilateral thyroid lobe total resection and bilateral IV lymph node dissection. Medullary thyroid cancer was confirmed by intraoperative frozen pathology in the right lobe of thyroid gland and papillary thyroid microcarcinoma in the left lobe of thyroid gland. No tumor recurrence and metastasis were found after 3-months follow-up.
Female ; Humans ; Middle Aged ; Thyroid Neoplasms ; diagnosis ; pathology ; surgery

Objective In order to improve cirrhotic liver management,each aspect of the liver's complex blood flow must be understood.This study investigates the protective effect of portal vein occlusion,with hepatic artery preservation,on cirrhotic liver after ischemia and reperfusion.Methods Carbon tetrachlorideand induced cirrhotic rats and normal rats were randomly assigned into 4 groups:normal sham operation (N-SO),cirrotic sham operation (C-SO),portal triad clamping (PTC),and portal vein clamping without hepatic artery inflow control (PVC).During the occlusion,the total 3-minute blood loss from the liver surface cut was weighed.At 1,6,and 24 hours post reperfusion,the serum alapine amino transferas (ALT),the adenosine triphosphate (ATP) of liver tissue,the malonolialdehgde (MDA) of liver tissue,and the morphological changes were evaluated.Result The amount of hemorrhage between the groups ranked as follows:PTC ＜ PVC ＜ N-SO ＜ C-SO (P＜0.05).At 1,6,and 24 hours post reperfusion.the ALT and MDA levels of the groups ranked as follows:PTC ＞ PVC ＞ C-SO ＞ N-SO (P＜0.05).Additionally,each group's ATP level ranked as follows:PTC ＜ PVC ＜ C-SO ＜ N-SO (P＜0.05).With histopathological examination,the hepatic injuries of the PTC and PVC group were more severe than those of the C-SO group,especially in the PTC group.Conclusion Therefore,the technique of portal vein clamping and hepatic artery inflow control can reduce the ischemic reperfusion injury of the cirrhotic rats' liver.

Objective To investigate the influence of double-effect activation of cholinergic antiinflammatory pathway on the liver injury and inflammatory response in obstructive jaundice rats by applying cholinesterase inhibitor and cholinergic M receptor blocker to activate alpha 7 nicotinic acetylcholine receptor.Methods 22 adult male Wistar rats were randomly assigned into three groups:sham operation (SO) group (n=6),bile duct ligation (BDL) induced obstructive jaundice with (BDL treatment group) or without treatment (BDL control group) (n=8 each).The medicine treatment group was given anisodamine (25 mg/kg) and neostigmine (25 μg/kg) daily via intraperitoneal injection after surgery,the control group was given equal amount of normal saline.The body weights of rats in each group were measured every other day.After 12 days,the rats were killed,and the pathological changes of liver injury,liver function and the expression levels of proinflammatory cytokines in the serum and liver tissue were observed.Results The body weight of BDL rats was significantly lower than the SO group rats,and the growth rate of BDL treatment group rats was the same as the rats in BDL control group 3 days after the starting of treatment.The AST,ALT,bilirubin and gamma-GT levels of BDL control and treatment groups were significantly higher than the SO group (P＜0.05),but there was no significant difference between BDL control and treatment groups.The serum albumin level of BDL treatment group was obviously higher than that of BDL control group,but the pathological liver injury was significantly slighter.The gene expression levels of TNF-alpha,IL-1 beta and IL-6 in the liver tissue were significantly higher in BDL groups than SO group (P＜0.05),but BDL treatment group was significantly lower than BDL control group (P＜0.05).In addition the serum TNF-alpha and IL-1 beta concentrations of BDL treatment group and control group were significantly higher than the SO group (P＜0.05),but the BDL treatment group was obviously lower than that BDL control group (P＜0.05).Conclusion The combine application of cholinesterase inhibitor and cholinergic M receptor blocker to activate the cholinergic anti-inflammatory pathway can significantly inhibit the obstructive jaundice induced proinflammatory gene expression and liver injury.

Objective To investigate the effects of triptolide on proliferation,apoptosis and the changes of Ski,Smad3,Smad7 and collagen type I(ColI) in cultured rat mesangial cells induced by transforming growth factor (TGF)-β1.Methods Cultured HBZY-1 rat mesangial cells were divided into 5 groups:(1)normal control group; (2)TGF-β1 group (10 μg/L); (3)-(5)triptolide (0.4,2,10 μg/L)+TGF-β1 (10 μg/L) groups.The cell proliferation was detected by MTT.Apoptosis of mesangial cells was detected by TUNEL assay.The expressions of Ski,Smad3,Smad7 mRNA were examined by real-time quantitative PCR.The expressions of Ski,Smad3,Smad7 and ColI protein were detected by Western blotting.The localizations of Ski and Smad3 protein were detected by laser confocal fluorescence microscope.Results Compared with the normal control,TGF-β1 (10 μg/L) significantly stimulated mesangial cells proliferation,while decreased apoptosis.The mRNA and protein expressions of Ski,Smad7,Smad3 and ColI protein expression in TGF-β1 group were increased (P ＞0.05).In comparison with TGF-β1 group,triptolide could significantly inhibit TGF-β1-induced mesangial cells proliferation in dose-dependent manner,and promote the apoptosis of mesangial cells.In TGF-β1 group,mRNA and protein expresscons of Ski and Smad7 were increased (P＜0.05),Smad3 mRNA and protein were decreased (P ＞0.05),and ColI protein was decreased (P＜0.01).In comparison with TGF-β1 group,fluorescence intensity of Ski,Smad3 proteins was significantly increased in cytoplasm,while decreased in nucleus.Conclusions Triptolide can inhibit TGF-β1-induced mesangial cells proliferation through regulating the expressions of Ski,Smad7 mRNA and protein,inhibiting Ski.Smad7 translocation to the nucleus,and down-regulating Smad3 mRNA and protein expression.Triptolide can promote apoptosis of mesangial cells.

Objective To evaluate the effects of drug-eluting stents (DES) versus bare-metal stents (BMS) on clinical outcomes in patients with acute ST-segment elevation myocardial infarction (ASTEMI) receiving primary percutaneous coronary intervention (PPCI). Methods The 217patients with ASTEMI receiving PPCI from Jan. 2005 to Dec. 2007 were enrolled in this study. And they were divided into two groups: DES group (n=92) and BMS group (n=125). The baseline characteristics including age, gender, angiographic characteristics, stents characteristics, Killip classification, cardiac troponin I(CTnI)levels, left ventricular ejection fraction(LVEF), hemoglobin levels, hypertension, diabetes, hyperlipidemia, obesity and smoking of the two groups were collected.Clinical follow-up end point were major adverse cardiac event(MACE)including death, acute myocardial infarction, stent thrombosis and stent restenosis. Clinical follow-up duration was(16.8±11.3) months (6-38 months). Results The average age (years), rate of Killip classification (class 2, 3, 4), average diameter (mm) of stent were significantly higher in BMS group than in DES group(64.6±11.9 vs. 61.2±11.8, t=2.09, P=0.037;25.9% vs. 12.2%, χ2=5.53, P=0.019;3.07±0.38 vs. 2.91±0. 40, t=2.78, P=0.006). And the average LVEF (%) was significantly lower in BMS group than in DES group (55.4±11.9 vs. 60.3±12.8, t= -2.57, P=0.011). The average length (mm) of stent, rate of stent post dilatation and diabetes were significantly higher in DES group than inBMSgroup (32.8±16.2 vs. 26.2±11.2, t=-3.54, P=0.001;45.7% vs. 21.6%, χ2=13.85, P=0. 000;28.2% vs. 16.0%, χ2=4.77, P=0.030). MACE occurred in 36 patients during clinical follow-up, 6 in DES group and 30 in BMS group. Incidence of MACE was significantly lower in DES group than in BMS group(6.5% vs. 24.0%, χ2=11.70, P＜0.01). Conclusions Using DES in ASTEMI patients is safe and may improve clinical outcomes by reducing incidence of MACE compared with BMS.