Objective To Investigate the mechanism of augmenter of liver regeneration (ALR) in promoting proliferation of damaged hepatocyte. Methods The inhibitory effects of ALR on the expression of TGF-?_1 in hepatic stellate cell and Kupffer cell were studied by immunohistochemistry and Western blot. The effects of hepatic stellate cell (Ito cell) conditioned medium (ICCM+) and Kupffer cell conditioned medium (KCCM+) prepared by treatment of using augmenter of liver regeneration (ALR) on damaged hepatocytes proliferation were studied by MTT. The antagonistical role of ALR on TGF-?_1, which inhibited damaged hepatocyte proliferation was investigated by MTT determination. Results Immunoreactive positive signal of TGF-?_1 in hepatic stellate cell and Kupffer cell stimulated by ALR were decreased. Immunolabeling of TGF-?_1 in hepatic stellate cell stimulated by ALR was weakened. The proliferation of damaged hepatocytes was increased significantly by administration of ICCM and KCCM. ALR could reverse the inhibitory role of TGF-?_1 on the proliferation of damaged hepatocyte. Conclusion ALR can promote proliferation of injured hepatocyte indirectly by inhibiting expression of TGF-?_1 in hepatic stellate cell and Kupffer cell.

Objective To approach the role of Kupffer cell (KC) in regulation of liver regeneration after partial hepatectomy (PH). Methods Condition medium of kupffer cells (KCCM) was prepared and the role of KCCM in promoting rat primary hepatocytes proliferation was observed by MTT method. The PH animal model was reproduced in mice and the rate of liver regeneration was measured and the expression features of TNF-?, TGF-? and TGF-?_1 were determined by immunohistochemical methods after PH with KC depletion. Results KCCM could promote primary hepatocyte proliferation significantly (A=0.746?0.06) compared with control (A=0.536?0.06, P

Objective To investigate the effect of RNA interference connective tissue growth factor (CTGF)expression on Kulclffer cells(KC)induced activation of hepatic stellate cells(HSC).Methods Rat CTGF RNA interference vector Psilencer 3.1H1-Neo-CTGF was constructed and identified.HSC cell line rHSC-99 cells were divided into three groups,group A served as control,group B transfected with vector without CTGF interference.group C was RNA interferenee CTGF expression of HSC.RT-PCR was used to measure the expression of CTGF in HSC.Rat Kur)ffer cells were isolated and identified.and cocultured with HSC in the 3 groups respectively.MTT assay was used to evaluate the proliferation of HSC.RT-PCR was used to measure the expression of TGF-β1 and precollagen type I in HSC.Western blot was used to measure the expression of TGF-β1 in HSC.ELISA was used to detect the production of precollagen type I protein.Immunofluorescence was used to detect the expression of ot-smooth muscle actin(α-SMA)in HSC.Resuits After CTGF RNA interference vector transfection.CTGF expression of HSC decreased by 22%(P＜0.01).The yield rate of Kupffer cell was 5×107 and the cell viability exceeded 98%.In the HSC and KC co-culture system.the proliferation and activation of HSC were inhibited while RNA interferenee CTGF of HSC.As compared with control,HSC proliferation decreased by 29%(P＜0.01).Precollagen type I and ot-SMA expression decreased by 38%(P＜0.01).Production of precollagen type I protein in culture medium decreased bv 48%(P＜0.01).Conclusions Blockade CTGF expression of HSC inhibits KC induced activation of HSC.

AIM: To investigate the mechanisms of augmenter of liver regeneration (ALR) in promoting damaged hepatocyte proliferation.METHODS: The effects of Kupffer cell condition medium (KCCM+) stimulated by ALR on damaged hepatocyte proliferation were studied by MTT. The localization of ALR binding to Kupffer cell membrane and in intact rat liver was studied by immunohistochemistry. The IL-6 expression in Kupffer cells stimulated with ALR was observed by immunohistochemistry. RESULTS: The proliferation of damaged hepatocytes stimulated with KCCM+ was increased significantly. ALR immunostaining particles in plasm of hepatocyte were found in intact liver. The rough immunostaining particles of ALR were seen on the surface of Kupffer cell membrane. Immunostaining particles of IL-6 in Kupffer cells induced by ALR increased. CONCLUSION: ALR promotes proliferation of damaged hepatocytes indirectly by stimulating Kupffer cells.

BACKGROUNDOne of mechanisms of carcinogenesis is suppression of cell apoptosis which leads to accumulation of aberrant cells. The aim of this study is to investigate cell apoptosis and COX-2 protein expression in non-small cell lung cancer (NSCLC).

METHODSCell apoptosis, expression of COX-2 and microvessel density (MVD) were detcted in 111 NSCLC samples by TdT-mediated dUTP nick end labeling (TUNEL) technique and immunohistochemical staining.

RESULTSThe positive rate of COX-2 protein expression was 67.6% (75/111), and there were 53 patients with high level cell apoptosis (47.7%). Expression of COX-2 protien was significantly related to TNM stages (P=0.025) and lymph node metastasis (P=0.018). The MVD in NSCLC tissues with positive COX-2 expression was significantly higher than that in negative expression ones (P=0.000). COX model showed that lymph node metastasis (P=0.006) and positive expression of COX-2 protein (P=0.000) were independent prognostic factors of NSCLC.

CONCLUSIONSThe expression of COX-2 protein may suppress cell apoptosis of tumor, and it may serve as a potential marker of prognosis for NSCLC.

OBJECTIVETo investigate the relationship of telomerase genes and the malignant transformation of atypical mammary ductal hyperplasia.

METHODSTelomerase genes hTR and hTRT in 50 cases of mammary hyperplasia (the cases included 6 benign hyperplasia, 9 mild atypical hyperplasia, 12 medium atypical hyperplasia, 23 severe atypical hyperplasia) and 26 cases of breast carcinoma were detected by in situ hybridization.

RESULTSThe expression of hTR and hTRT mRNA were weak or negative in benign hyperplasia (1/6, 0), weaker in mild-moderate atypical hyperplasia (2/9, 1/9, 4/12, and 3/12), strong in severe atypical hyperplasia (14/23, 60.9% and 12/23, 52.1%), while very strong expression (23/26, 88.5% and 21/25, 80.8%) in carcinoma of the breast. The difference between mild-moderate atypical hyperplasia, invasive ductal carcinoma and severe atypical hyperplasia was significant (P < 0.05) and the difference between severe atypital hyperplasia and intraductal carcinoma was not significant (P > 0.05).

CONCLUSIONSTelmerase genes (hTR, hTRT) expression is closely related to the malignant transformation of atypical hyperplasia. The reactivated telomerase may play a crucial role in the development of breast cancer.

Breast Neoplasms ; enzymology ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Gene Expression ; Humans ; RNA, Messenger ; Telomerase ; genetics

OBJECTIVETo investigate telomerase gene expression in precancerous mammary lesion, such as atypical ductal hyperplasia and breast cancer and to study the relationship between expression and malignant transformation.

METHODSExpression of human telomerase genes (hTR) and human reverse transcriptase gene (hTRT) in 76 cases of mammary tissue was evaluated using in situ hybridization and included 50 cases of mammary hyperplasia, 6 of which were benign hyperplasia, 9 were mild atypical hyperplasia, 12 were moderate atypical hyperplasia, 23 were severe atypical hyperplasia and 26 were mammary cancer.

RESULTSThe expressions of hTR and hTRT mRNA were much weaker or negative in benign hyperplasia (16.6%, 0), weak to mild moderate in atypical hyperplasia (22.2%, 11.1%, 33.3%, 25.0%), strong in severe atypical hyperplasia (60.9%, 52.1%), and significantly strong in mammary cancer (88.5%, 80.8%). The difference between mild-moderate atypical hyperplasia, invasive ductal carcinoma and severe atypical hyperplasia was significant (P < 0.05) and the difference between severe atypical hyperplasia and intraductal carcinoma was not significant (P > 0.05).

CONCLUSIONTelomerase genes (hTR and hTRT) expressions are related to the transformation of atypical hyperplasia. Activated telomerase may play a role in mammary cancer development.

Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; pathology ; DNA-Binding Proteins ; Female ; Gene Expression ; Humans ; Precancerous Conditions ; genetics ; pathology ; RNA ; genetics ; physiology ; RNA, Messenger ; analysis ; Telomerase ; genetics ; physiology

Objective To establish a rapid and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)analysis method for determination of unsymmetrical dimethylhydrazine(UDMH)contents in rat plasma and investigate the toxicokinetic characteristics of UDMH in rats.Methods Twenty-two SD rats were divided into the intravenous injection of 10 mg/kg dose group(4 females)and intragastric administration groups(low,medium and high dose,with 6 rats in each group,half males and half females).The rats were given 10mg/kg by intravenous administration and 10 mg/kg,30 mg/kg,and 90 mg/kg single dose of UDMH by gavage.Blood samples were collected from the orbital venous plexus at 0 hour before administration and at different time points after administration.The plasma samples were extracted with protein precipitation and derivatization before being analyzed using the LC-MS/MS method.Separation was carried out on a ZORBAX column(4.6mm×75mm,3.5 μm),with a mobile phase composed of 0.3％acetonitrile/formic acid at a flow rate of 1 mL/min.Propranolol was used as the internal standard.An electrospray ionization(Turbo Ionspray)source was applied and the mass spectrometer was operated in a positive MRM mode.Quantitative analysis showed that the ionization source unsymmetrical dimethylhydrazine and propranolol was at m/z:192.0→148.1,m/z:260.2→116.1,respectively.The toxicokinetic parameters were analyzed with the DAS 2.1 software.Results Quantification of UDMH exhibited a good linearity within the concentration range of 50-50000 ng/mL,with a linear correlation coefficient greater than 0.9900 and a lower limit of quantification of 50 ng/mL.The average recovery rate of UDMH was 98.1％,compared with 100.5％for the internal standard propranolol hydrochloride.The inter batch precision of standard curve samples ranged from 0.7％to 6.3％,and the relative error was between-7.1％and 6.2％.The inter batch and intra batch precision of quality control samples ranged from1.8％to 19.8％,and the relative error from-9.8％to 0.2％.The main pharmacokinetic parameters of UDMH in rats exposed to 10 mg/kg,30 mg/kg,and 90 mg/kg gavage were UC(0-t):(7624.99±2569.31),(34284.04±6657.15),(84720.88±22354.80)μg/L·h,t1/2:(0.07±0.15),(2.24±1.45),(3.04±0.90)h,Tmax:(0.75±0.27),(0.51±0.29),(0.29±0.10)h,Cmax:(4454.14±1329.45),(19442.45±9121.07),(32334.35±9882.41)μg/L,F:(77.34±26.06)％,(115.92±22.51)％,(95.48±25.19)％.Conclusion The LC-MS/MS method is highly accurate and specific,and is suitable for the toxicokinetic study of UDMH in rats.Single gavage administration of UDMH results in absorption and elimination saturation at a high dose.This study provides data for toxicological studies related to UDMH.

Objective To assess the efficacy and side effects of intravesical instillation of BCG after transurethral resection of the bladder tumor (TURBT) in non-muscle invasive bladder cancer (NMIBC) patients.Methods The clinical data of patients treated with BCG 120 mg per course induced perfusion or more after TURBT from December 2013 to October 2016 in 18 hospitals of northeast China region,were analyzed retrospectively.The first part,data of 106 patients with moderate,high-risk NMIBC were collected.A total of 83 patients were male,while the other 23 patients were female.The average age was 66.7 years old.The clinical staging were T1 in 86(81.1％) cases,Ta in 20(18.9％) cases and carcinoma in situ in 6 (5.7％) patients.Intravesical instillation of BCG was executed after transurethral resection of the bladder tumor.The incidence rate of recurrence and progression during more than 6 months' follow-up time were observed.Multivariate analyses were done by using logistic analysis and Cox proportional hazards regression model with Kaplan-Meier method.The second part,treatment compliance of 276 patients with bladder cancer,including moderate/high-risk NMIBC in 263 cases,moderate/high-risk NMIBC followed with renal pelvis/ureteral carcinoma in 8 cases were and moderate/high-risk NMIBC with renal pelvis/ureteral carcinoma in 5 cases who treated with BCG after the surgeries,were observed.Patients consisted of 211 males and 65 females with average age of 68.3 years.Results With a median follow-up of 12 months,9 (8.5％) patients experienced tumor recurrence and 2 (1.9％) patients were found progression in the first part.The one-year cancer free recurrence rate of the patients was 91.5％.Statistically significant prognostic factors for recurrence identified by multivariable analyses were prior recurrence of the tumors (OR =3.214,95％CI0.804-12.845,P =0.099).In the second port,an incidence rate of adverse effects was 64.1％ (177/276).The Ⅲ/Ⅳ degree complications were occurred in 11 patients and satisfactory outcomes achieved with active treatment.A total of 36 patients withdrawal with the major causes were recurrence and progression of bladder tumor in 12 cases (4.4 ％),9 cases (3.3 ％) with economic reasons and 11 cases (4.0％) with serious complications.Conclusions NMIBC patients treated with intravesical BCG therapy have approving cancer free recurrence rates and acceptable adverse effects.Prior recurrence may be prognostic factor of recurrence after intravesical BCG therapy.

OBJECTIVE: To investigate the predictive value of pancreatitis activity scoring system (PASS) combined with Neutrophil to lymphocyte ratio (NLR) and C-reactive protein (CRP) for infected pancreatic necrosis (IPN) in patients with severe acute pancreatitis (SAP). METHODS: Clinical data of SAP patients admitted to the First Affiliated Hospital of Zhengzhou University from January 2020 to January 2023 were retrospectively collected, including basic information, vital signs at admission, first laboratory indexes within 48 hours of admission. The PASS scores at admission and 24, 48 and 72 hours after admission were calculated. According to the diagnostic criteria of IPN, the patients were divided into the non-IPN group and the IPN group, and the independent risk factors of SAP complicating IPN were determined by using univariate analysis and multifactorial Logistic regression. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of NLR, CRP, and PASS score, alone and in combination for IPN in patients with SAP. RESULTS: A total of 149 SAP patients were enrolled, including 102 in the non-IPN group and 47 in the IPN group. The differences in PASS score at each time point, NLR, CRP, procalcitonin (PCT), blood urea nitrogen, blood chloride, and days of hospitalization between the two groups were statistically significant. Multifactorial Logistic regression analysis showed that 72 hours admission PASS score [odds ratio (OR) = 1.034, 95% confidence interval (95%CI) was 1.005-1.065, P = 0.022], NLR (OR = 1.284, 95%CI was 1.139-1.447, P = 0.000), and CRP (OR = 1.015, 95%CI was 1.006-1.023, P = 0.001) were independent risk factors for IPN in patients with SAP. ROC curve analysis showed that the area under the ROC curve (AUC) of the PASS score at 72 hours of admission, NLR, and CRP alone in predicting IPN in SAP patients were 0.828, 0.771, and 0.701, respectively. The AUC of NLR combined with CRP, PASS combined with NLR, and PASS combined with CRP were 0.818, 0.895, and 0.874, respectively. The combination of PASS score at 72 hours after admission, NLR, and CRP had a better predictive ability for IPN in patients with SAP (AUC = 0.922, 95%CI was 0.877-0.967), and the sensitivity was 72.3% when the cut-off value was 0.539. CONCLUSIONS The predictive value of the PASS score at 72 hours after admission, NLR and CRP in combination for IPN in SAP patients is better than that of the combination of each two and individual detection and has better test efficacy.
Humans ; Pancreatitis, Acute Necrotizing/diagnosis* ; C-Reactive Protein/metabolism* ; Acute Disease ; Neutrophils/metabolism* ; Retrospective Studies ; ROC Curve ; Lymphocytes ; Prognosis