The pancreatic enzyme-II type collagenase digestion method was adopted for primary culture of osteoblasts, inoculation and passage. They were identified by alkaline phosphatase dye-liquor. N-butanol extract fractions from different processed products of Cibotium barometz were prepared. The above osteoblasts were jointly cultured with protocatechuic acid, protocatechuic aldehyde, kojic acid and the mixed control liquid of the above three substances, and their proliferation was detected by CCK-8. Various n-butanol extract fractions from different processed products of C. barometz showed a significant proliferative effect on osteoblasts in the order of the wined > the heated > the salted > the sand-heated and wined system > the alcohol-processed > the steamed > the crude. The q test showed no significant difference among sand-heated, alcohol-processed and steamed C. barometz, no significant difference between heated and salted C. barometz. Various control substances also showed a certain proliferative effect on osteoblasts in the order of the mixed control > protocatechuic aldehyde > protocatechuic acid > kojic acid. The q test showed no significant difference between protocatechuic aldehyde and protocatechuic acid. All of n-butanol extract fractions from different processed products of C. barometz showed a significant effect on osteoblast proliferation, of which wined C. barometz showed the best effect. All of phenolic compounds such as protocatechuic aldehyde, protocatechuic acid and kojic acid showed a significant proliferative effect on osteoblasts.
Animals ; Cell Proliferation ; drug effects ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Osteoblasts ; cytology ; drug effects ; Pteridophyta ; chemistry ; Rats ; Rats, Sprague-Dawley

Objective To evaluate ultrasound-guided methylene blue dyeing for radical liver segmental resection.Methods Liver segmental resection with uhrasound-guide methylene blue dyeing (UMD-SR) was performed in 16 cases, results were compared with 16 conventional liver segmental resection (CSR) retrospectively.Results All the operations under uhrasound-guided methylene blue dyeing were successfully carried out, among them, reverse dyeing was used in cases with segment Ⅳ, Ⅴ, Ⅷ resection.The blood loss in UMD-SR group was much less than CSR group(t =3.011 ,P =0.009) , at the cost of a longer operation time (t =5.423,P =0.000 07).There was no difference in the mortality and morbidity rates between two groups.Tumor recurrence rate was 6.25％ in UMD-SR group and 18.75％ in CSR group (x2 =0.133,P =0.285).Conclusions Ultrasound-guided methylene blue dyeing liver segmental resection can reduce the blood loss during operation, improve the safety of hepatectomy in case of hepatic carcinoma.

Objective To investigate the CT and MRI findings of primary hepatic neuroendocrine carcinomas.Methods Imaging findings of 6 patients with pathologically confirmed hepatic neuroendocrine carcinomas were retrospectively analyzed.Four patients underwent plain CT and contrast enhanced CT examinations.The other 2 patients underwent plain MRI and contrast enhanced MRI.Results One out of the 6 cases manifested multicentric tumor which appeared as one large tumor surrounded with multiple small nodules peripherally,and the tumors of the other 5 cases were solitary.On plain CT images,all lesions were well-defined and hypoattenuating with central areas of even lower density except punctuate calcifications at tumor periphery in 1 case.On pre-contrast MRI,the cases manifested heterogeneous low signal intensity on T1 wI and slightly high intensity on T2 WI.On dynamic contrast enhanced CT and MRI,Tumors manifested slight to mild enhancement in arterial phase,slight enhancement in portal venous phase or delayed phase with no enhancement in the center,accompanied by dislocation of surrounding vessels.There Was no lymphadenopathy in peritoneal cavity and retroperitoniurm. Conclusion CT and MRI scanning can demonstrate specific features of primary hepatic neuroendocrine carcinoma,which may help the diagnosis and differential diagnosis of the tumor.

Objective To investigate the association between catechol-O-methyltransferase (COMT) Val158Met polymorphism and the memery function of schizophrenia treated with Aripiprazole.Methods Schizophrenic patients were diagnosed in according to Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV).A total of 78 Chinese Han subjects were involved in this study.The polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) was used to identify COMT genotypes.The Chinese revised version of Wechsler Memory Scale (WMS-RC) was used twice to evaluate the memory function of patients treated with Aripiprazole alone before and after drug treatment for 8 weeks.ANOVA,rank sum test and t-test were used to statistical analysis.Results There was statistical difference of cumulative memory among the three groups (F=12.371,P=0.002),and Met/Met genotype showed much poorer cumulative memory than Val/Met and Val/Val genotypes (Z=-3.168,-2.641 ; P=0.002,0.008,respectively) Memory function was significantly improved after patients been treated with Aripiprazole except the associative memory(P＜0.05).Met/Met genotype showed more improvement of cumulative memory than other genotypes after Aripiprazole treatment (t=4.203,P＜0.01 ; t=3.024,P ＜0.01,respectively).Conclusions There may be a relationship between COMT Val158Met polymorphism and the memory function of schizophrenia in Aripiprazole treatment.Met/Met genotype of schizophrenia showed worst cumulative memory but best responsiveness in Aripiprazole treatment.

We assessed the effects of pulsatile flow shear stress on the gene expression profiles of human umbilical artery smooth muscle cells (HUASMCs) in vitro using the Express Chip DNA microarray method and investigated the difference between pulsatile and steady shear stress on differentially expressed genes of HUASMCs. In a modified pulsatile flow chamber system, HUASMCs were exposed to pulsatile and steady fluid shear stress (5.52 dyne/cm2) for 6 h respectively, and normal static cultured HUASMCs were selected as a control. The total cellular RNA was extracted by TRIzol Reagent (Life Technologies, Inc) according to the manufacturer's manual. Conversion of mRNA to single strand cDNA and double strand cDNA template was synthesized by Reverse Transcription from the total RNA. cRNA probe was transcribed with biotin labeling. After hybridization of probe with microarray, the binding of streptavidin to biotin was performed and amplified with the first antibody and further amplified with Cy3-conjugated second antibody. Then detection of Cy3 dye was carried out with ScanArray 5000. The results showed that a total of 1,330 genes revealed differential expression in HUASMCs exposed on pulsatile shear stress (5.52 dyne/cm2, 6 h); however, 2,676 genes revealed differential expression in HUASMCs exposed on steady shear stress. Comparsion of HUASMCs exposed to pulsatile with the HUASMCs exposed to steady shear stress showed there were 2,297 genes revealing differential expression. The transcriptional profile of fluidally induced genes in HUASMCs suggested a different response to pulsatile and steady shear stress.
Cells, Cultured ; Gene Expression Profiling ; Humans ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Pulsatile Flow ; RNA, Messenger ; biosynthesis ; genetics ; Stress, Mechanical ; Umbilical Arteries ; cytology

BACKGROUND: Increasing evidence shows that at least 155 genetic polymorphisms are associated with aerobic performance and elite endurance athlete status. OBJECTIVE: To study the association of α-actin 3 (ACTN3) gene, nuclear respiratory factor 2 (NRF2), β2 adrenergic receptor (ADRB2) gene and peroxisome proliferative activated receptor gamma coactivator 1 alpha (PPARGC1A) polymorphic loci with aerobic performance of rowing athletes and their interaction effect, thereby providing basis for understanding the mechanism of genetic polymorphisms acting on endurance athlete status. METHODS: A case-control experiment was designed to analyze the distribution characteristics of four gene polymorphism loci in 15 excellent rowing athletes and 50 common college freshmen. The association of four polymorphic loci with the aerobic performance-related indexes was analyzed using total genotype scores. RESULTS AND CONCLUSION: The preponderant polymorphism locus distributions of ACTN3, PPARGC1A and NRF2 in the athletes group were higher than those in the control group, and the NRF2 gene loci showed significant difference between two groups. In the athletes group, the mean values of VO2 maxwas significantly different among three genotypes. That is to say, these three genetic polymorphisms may be the biomarkers to predict the elite endurance athlete status, but the mechanism needs to be studied in depth.

Adult rats chronic unpredictable stress model of depression (CUS) was adopted to elucidate the antidepressant pharmacological activity and related neurogenesis protective effect of the total flavonoids extract (licorice flavonoids, LF) from the Glycyrrhiza uralensis Fisch. cultivated locally in Ningxia. The rats were exposed to 9 kinds of unpredictable sequence of stressors and were given flavonoids (300 mg x kg(-1), 100 mg x kg(-1) and 30 mg x kg(-1)) for 28 days. The antidepressant effect was elucidated by open field test, forced swimming test and tail suspension test. The level of serum corticosterone was detected by radioimmunoassay. 5'-Bromo-2'-deoxyuridine (BrdU) labeling experiments was employed to study the neurogenesis protective activities. The flavonoids can increase the sum of line crosses and number of rears, and decrease the number of fecal boli produced in the open field test of the CUS rats. Also the flavonoids can decrease the immobility time in forced swim test as well as in the tail suspension test. In addition, the flavonoids (300 mg x kg(-1)) can decrease the serum corticosterone level of the CUS rats, and increase the number of the new born BrdU positive progenitor cells at the subgranular zone (SGZ) of dentate gyrus (DG) region in hippocampus. The results demonstrated that the total flavonoids extract from the cultivated Glycyrrhiza uralensis Fisch. could produce the anti-depressive effect on chronic unpredictable stress of depression model rats and its mechanism may be associated with its neurogenesis protective effect.

OBJECTIVETo investigate the protective effect and action mechanism of petroleum ether extracts from Saussurea involucrate on brain tissues of hypoxia rats under constant pressure and closed conditions.

METHODThe PESI dosage-dependent experiment for hypoxia rats was conducted under constant pressure and closed conditions by intraperitoneally injecting 125, 250, 500 mg x kg(-1) to finalize that the optimum dosage is the high dose of PESI. Afterwards, 90 Wistar rats were randomly divided into the hypoxic model group, the acetazolamide 250 mg x kg(-1) group and the PESI high dose group. Each group was further divided into three subgroups according to different hypoxia times, with 10 rats in each subgroup. Under the same hypoxia and administration conditions, the rats were sacrificed after 0, 3, 6 h respectively. Their brain samples were collected for common pathological observation and immunohistochemical staining of HIF-1alpha. Real-time RT-PCR was used to detect HIF-1alpha, EPO, HO-1 and Caspase-3 gene expressions. And the Western blot assay was adopted to detect HIF-1alpha protein expression.

RESULTThe brain tissues of the hypoxia model group were severely damaged with the increase in the hypoxia time. The acetazolamide group and the PESI high does group were damaged in a much lower degree. According to the gene expression and the Western blot assay, high dose of PESI could inhibit HIF-1alpha expression. According to the pure gene expression test, high dose of PESI could increase EPO and HO-1 mRNA expressions, but inhibit Caspase-3 mRNA expression.

CONCLUSIONPESI's protective mechanism for brain tissues of hypoxia rats under constant pressure and closed conditions may be related to its effects in inhibiting HIF-1alpha expression, increasing EPO expression and resisting cell apoptosis.

Alkanes ; chemistry ; Animals ; Brain ; cytology ; drug effects ; metabolism ; Caspase 3 ; genetics ; Cell Hypoxia ; drug effects ; Cytoprotection ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Erythropoietin ; genetics ; Gene Expression Regulation, Enzymologic ; drug effects ; Heme Oxygenase-1 ; genetics ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Male ; Rats ; Rats, Wistar ; Saussurea ; chemistry

OBJECTIVETo investigate the effect of Biejiajian Pills on Wnt signal pathway and its inhibitory gene (DKK-1 and FrpHe) expressions and explore the mechanism underlying the action of Biejiajian Pills to suppress the invasiveness of hepatocellular carcinoma.

METHODSTwenty-four Wistar rats were randomized equally into 3 groups for gavage of normal saline and Biejiajian Pills at 20- and 10-fold clinical doses for 3 days. Blood samples were then collected from the rats, and the serum was separated and added in HepG2 cell cultures. After 48 h of culture, the cells were collected to determine the cellular content of β-catenin protein using flow cytometry and detect DKK-1 and FrpHe mRNA expressions using qRT-PCR.

RESULTSHepG2 cells cultured in the presence of sera from rats fed with Biejiajian Pills showed significantly lowered β-catenin protein expression and obvious down-regulation of DKK-1 mRNA expression, and the effect was correlated with the doses of the drug administered. The expression of FrpHe mRNA showed no significant differences between the 3 groups.

CONCLUSIONSBiejiajian Pills can effectively inhibit the invasiveness and migration of hepatocellular carcinoma cells, which is closely related to decreased expressions of β-catenin and DKK-1 to cause block of the Wnt/β-catenin signal pathway.

Animals ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Male ; Proto-Oncogene Proteins ; metabolism ; Rats ; Rats, Wistar ; Wnt Proteins ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

Objective:To investigate the clinical efficacy of the combination therapy of lenvatinib and programmed death-1 (PD-1) antibodies in unresectable or advanced hepatocellular carcinoma (HCC).Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of 59 patients with unresectable or advanced HCC who were admitted to Zhongshan Hospital of Fudan University from September 2018 to January 2020 were collected. There were 54 males and 5 females, aged from 25 to 73 years, with a median age of 52 years. All 59 patients underwent combination therapy with lenvatinib and PD-1 antibodies including 43 cases undergoing first-line therapy and 16 cases who cannot tolerate first-line therapy or with tumor progressed after first-line therapy undergoing second-line therapy. Observation indicators: (1) clinical efficacy; (2) adverse drug reactions and treatment; (3) follow-up and survival. Follow-up was performed using outpatient examination or telephone interview to detect tumor diameter of the target lesion, overall survival and progression free survival of patients up to December 2020. Measurement data with skewed distribution were expressed as M ( P25,P75) or M (range). Count data were represented as absolute numbers and (or) percentages. The Kaplan-Meier method was used to calculate the median duration of response (DoR), median overall survival time, median progression free survival time, survival rates and draw survival curves. Results:(1) Clinical efficacy: the objective response rate (ORR), complete response rate (CR), partial response rate (PR), stable disease rate (SD), progression disease rate (PD), time to response (TTR) and median DoR of 59 HCC patients were 37.3%(22/59), 11.9%(7/59), 25.4%(15/59), 37.3%(22/59), 25.4%(15/59), 2.6 months(2.1 months, 4.0 months), 6.3 months[95% confidence interval ( CI) as 2.2 to 10.5 months], respectively. The ORR, CR, PR, SD, PD and TTR of 43 HCC patients undergoing first-line therapy were 41.9%(18/43), 16.3%(7/43), 25.6%(11/43), 37.2%(16/43),20.9%(9/43), 2.2 months(2.0 months, 3.5 months), respectively. The median DoR of 43 patients undergoing first-line therapy was not reached. The ORR, CR, PR, SD, PD, TTR and median DoR of 16 HCC patients undergoing second-line therapy were 4/16, 0, 4/16, 6/16, 6/16, 3.8 months (3.6 months, 4.1 months), 4.2 months(95% CI as 2.0 to 6.3 months), respectively. Six of 59 HCC patients underwent R 0 resection due to tumor converting to resectable HCC with the conversion and resection rate of 10.2%(6/59). Among the 6 patients, 5 cases undergoing first-line treatment had the conversion and resection rates of 11.6% (5/43) and 1 case undergoing second-line treatment had the conversion and resection rates of 1/16, respectively. (2) Adverse drug reactions and treatment: 25 of 59 HCC patients underwent 3 to 4 grade adverse drug reactions with the incidence of 42.4%(25/59). Among the 25 patients, 10 cases including 5 cases undergoing first-line therapy and 5 cases undergoing second-line therapy had the level of gamma glutamyltransferase >5×upper limit of normal (ULN), 9 cases including 4 cases undergoing first-line therapy and 5 cases undergoing second-line therapy had the level of aspartate aminotransferase >5×ULN, 5 cases including 4 cases undergoing first-line therapy and 1 case undergoing second-line therapy occurred gastrointestinal hemorrhage, 4 cases undergoing first-line therapy had the level of white blood cell count <2.0×10 9/L, 4 cases including 1 case undergoing first-line therapy and 3 cases under-going second-line therapy had the level of total bilirubin >3×ULN, 3 cases undergoing first-line therapy had the level of neutrophil count <1.0×10 9/L, 3 cases including 2 cases undergoing first-line therapy and 1 case undergoing second-line therapy occurred ascites, 2 cases including 1 case undergoing first-line therapy and 1 case undergoing second-line therapy had the level of platelet count <50.0×10 9/L, 2 cases undergoing first-line therapy had the level of alanine aminotransferase >5×ULN, 2 cases undergoing first-line therapy occurred hyponatremia, 2 cases including 1 case undergoing first-line therapy and 1 case undergoing second-line therapy occurred pulmonary infection, 2 cases including 1 case undergoing first-line therapy and 1 case undergoing second-line therapy occurred type 1 diabetes, 1 case undergoing first-line therapy occurred hypokalemia, 1 case undergoing first-line therapy occurred myocarditis, 1 case undergoing first-line therapy occurred hypophysistis, 1 case undergoing first-line therapy occurred bullous dermatitis, 1 case undergoing first-line therapy occurred hypertension. Three of 59 HCC patients underwent 5 grade adverse drug reactions ,with the incidence of 5.1%(3/59), including 1 case undergoing first-line therapy with immune hepatitis, 1 case undergoing second-line therapy with immune pneumonia and 1 case undergoing second-line therapy with immune enteritis. Some of patients underwent multiple adverse drug reactions at the same time. Twenty five patients undergoing 3 to 4 grade adverse drug reactions were relieved with the treatment of drug reduction, drug withdrawal, symptomatic treatment or hormone therapy. Three patients undergoing 5 grade adverse drug reactions died after being treated with high-dose hormone shock and hepatoprotective treatment. (3) Follow-up and survival: all 59 patients were followed up for 1.5 to 25.2 months, with a median follow-up time of 13.3 months. Of them, patients undergoing first-line therapy were followed up for 1.9 to 25.2 months, with a median follow-up time of 13.5 months. During follow-up,20 cases undergoing first-line therapy died with the fatality rate of 46.5%(20/43). Patients undergoing second-line therapy were followed up for 1.5 to 24.4 months, with a median follow-up time of 10.8 months. During follow-up, 10 cases undergoing second-line therapy died with the fatality rate of 10/16. Up to the latest follow-up, the tumor diameter of the target lesion in all 59 patients, in patients undergoing first-line therapy and in patients undergoing second-line therapy was 75 mm(38 mm, 125 mm), 74 mm(36 mm, 116 mm), 84 mm(48 mm,150 mm), respectively. The ratio of tumor diameter of the target lesion at latest follow-up to tumor diameter of the target lesion at baseline were -9.05%(-27.3%, 19.7%), -16.1%(-28.8%, 13.6%), 13.2%(-24.7%, 23.5%) for all 59 patients, patients undergoing first-line therapy and patients undergoing second-line therapy, respectively. The median overall survival time and median progression free survival time of patients undergoing first-line therapy and patients undergoing second-line therapy were 17.1 months(95% CI as 11.0 to 23.2 months), 10.8 months(95% CI as 5.0 to 16.6 months) and 10.8 months(95% CI as 9.2 to 12.4 months), 3.0 months(95% CI as 1.6 to 4.4 months), respectively. Conclusion:For unresectable or advanced HCC, combination therapy with lenvatinib and PD-1 antibodies can obtain effective antitumor activity and less incidence of adverse drug reactions.