BACKGROUND:We have successfuly prepared digital coraline hydroxyapatite artificial bone scaffold in previous experiments, and it has been confirmed that it has the necessary physical and chemical properties of bone tissue engineering scaffolds. OBJECTIVE: To evaluate the sensitization of digital coraline hydroxyapatite artificial bone scaffold. METHODS:A total of 32 guinea pigs were randomly divided into saline group (negative control group, n=8), 5% formaldehyde group (positive control group,n=8), experimental A group (the mass ratio of 3:1,n=8), and experimental B group (the mass ratio of 4:1,n=8). Sensitization test at the maximal dosage was performed according toBiological Evaluation of Medical Devices-Part 10: Tests for Irritation and Delayed-Type Hypersensitivity, including intracutaneous induction, local induction, and provocation. Patch was removed after 24 and 48 hours, and the skin response was classified according to Magnusson and Kligman criteria. Patch was removed after 48 hours, and the skin was performed with biopsy, stained with hematoxylin-eosin, and observed under optical microscope. RESULTS AND CONCLUSION: Sensitization response was not tested in the negative control group, experimental A group and experimental B group at 24 and 48 hours after patch removal; however, moderate erythema was observed in the positive control group. Optical microscope demonstrated that spongiosis, edema, diffuse or perivascular mononuclear infiltration was not observed, and only a smal number of basicytes were seen in the experimental A and B groups. These findings indicate that the digital coral hydroxyapatite artificial bone scaffolds, with the mass ratio of 3:1 and 4:1, are biologicaly safe for sensitization.

A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

Objective:To explore whether the 15-minute retention rate (R15) of indocyanine green (ICG) in the indocyanine green excretion test and the effective hepatic blood flow (EHBF) can predict the occurrence of mild hepatic encephalopathy (MHE).Methods:Using the convenience sampling method, we collected clinic data from 153 patients diagnosed with liver cirrhosis or liver failure from June 2019 to December 2019 in the Third Affiliated Hospital of Sun Yet-sen University in Guangzhou. We screened the MHE patients with the number connect test-A and the digital symbol test, and analyzed the clinical data. By taking different values for R15 and EHBF as cut-off points, the significance of the two factors in predicting MHE is explored respectively.Results:The incidence of MHE was 38.56% (59/153). Single factor analysis showed that the difference of Child-Pugh grade between the MHE group and the non-MHE group was statistically significant ( χ2 value was 7.606, P<0.05), while the differences between cirrhosis and liver failure diagnosis, fasting blood glucose, and serum creatinine were not statistically significant ( P>0.05). When most points between 0.11 and 0.61 were selected as normal and abnormal cut-off points of R15, R15 had statistical significance ( P<0.05) and when R15 selected 0.18, it was most significant (Fisher exact test P=0.00024). When most points between 0.08 and 0.76 were selected as normal abnormal cut-off points of EHBF, EHBF had statistical significance ( P<0.05) and when EHBF selected 0.25, it was most significant (Fisher exact test P=0.00022). Through Logistic stepwise regression analysis, the risk factors for MHE were R15 and EHBF. The ROC curve was used to illustrate the predictive effects of two factors on MHE. Conclusions:The incidence of MHE in patients with cirrhosis or liver failure is high. When R15≥0.18 or EHBF≤0.25 L/min, R15 and EHBF in indocyanine green excretion test can better predict the occurrence of MHE than Child-Pugh classification, and can be help to evaluate and manage patients in time.

OBJECTIVETo investigate the role of Xuebijing injection(XBJI, traditional Chinese medicine), in inhibiting TLR4--NF-kappaB--IL-1beta pathway of myocardial hypoxia/reoxygenation in rats.

METHODSThirty six male SD rats (280 +/- 30) g were randomly divided into six groups (n = 6): normal group (N group), balanced perfusion group (BP group), model group (M group), low dose XBJI group (XBJI(L) group), middle dose XBJI group (XBJI(M) group), high dose XBJI group (XBJI(H) group). By Langendorff isolated heart perfusion device to establish the model of myocardial hypoxia/reoxygenation in rats. ELISA was used to detect the concentration of interleukin-1beta (IL-1beta); Western blot was used to detect the expression of nuclear factor kappa B p65 (NF-kappaB p65) protein and toll like receptor 4 (TLR4) protein; and RT-PCR to determine the expression of NF-kappaB p65 mRNA and TLR4 mRNA;To observe microstructure changes of hypoxia/reoxygenation myocardial by light microscopy.

RESULTSCompared with M group, the IL-1beta concentration, NF-kappaB p65 and TLR4 protein,NF-kappaB p65 and TLR4 mRNA of XBJIL group, XBJI(M) group, XBJI(H) group expression decreased in varying degrees,and decreased most obviously all in XBJI(M) group (P < 0.05, P < 0.01); Myocardical structural damage was serious in M group, and improved after treatment XBJI, the most obvious was the XBJI(M).

CONCLUSIONDifferent dose of XBJI can inhibit TLR4--NF-kappaB--IL-1beta signal transduction pathway and reduce several inflammatory reaction after myocardial hypoxia/reoxygenation injury, the 4 ml/100 ml of XBJI is the best.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Heart ; drug effects ; Inflammation ; Interleukin-1beta ; metabolism ; Male ; Myocardium ; pathology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

OBJECTIVETo evaluate the effect of Sapylin combined with intraperitoneal and systemic chemotherapy for advanced colon cancer following radical operation on local recurrence, hepatic metastasis, and overall survival rate.

METHODSFrom Jan. 2004 to Dec. 2005,132 patients with stage II or III colon carcinoma after radical operation were randomly divided into two groups: Sapylin combined with chemotherapy(Sapylin) group and the control group. Toxic reaction, local recurrence, hepatic metastasis, and overall survival rate between two groups were compared.

RESULTSBoth groups successfully completed the trial. There was no significant difference in toxic reaction between two groups, the recurrence and hepatic metastasis rate in Sapylin group were lower than those in the control group(9/60, 15.0% vs. 22/72, 30.6%; 11/60,18.3% vs. 22/72, 34.7%, respectively), which were statistically significant (P<0.05 respectively). The 3- year survival rate in Sapylin group was higher than that in control group(73.3% vs. 54.2%), which was statistically significant (P<0.05).

CONCLUSIONSapylin combined with intraperitoneal and systemic chemotherapy can effectively decrease local recurrence and hepatic metastasis,and improve the survival in patients with advanced colon cancer following radical operation.

Adult ; Aged ; Biological Products ; therapeutic use ; Chemotherapy, Adjuvant ; Colonic Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Postoperative Period ; Prospective Studies ; Streptococcus pyogenes ; Survival Rate

A comprehensively comparison of the chemical profiles between sun-drying BJ (NBJ) and sulfur-fumigated BJ (SBJ) was conducted by HPLC analysis and the discrepant peaks were identified or tentatively assigned by HPLC-ESI-MSn. A total of 32 chemical components were used for qualitative comparison. Meanwhile, a quantitative comparison of BJwere conducted by HPLC analysis and determining seven compounds from 3 NBJ and 3 SBJ samples dramatic chemical changes were found. After sulfur fumigation, the contents of flavonoids glycosides and phenolic acids were remarkably reduced, but the contents of flavonoids aglycones were significantly increased. Multivariate statistics, including principle component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) were used to investigate the potential damaging effect of sulfur-fumigating process. The PCA score plots showed six samples were clearly classified into the sun-drying and sulfur-fumigating groups. And according to VIP >1, the most important chemical markers were apigenin, luteolin and 3,5-dicaffeoylquninic acid which could be used to distinguish NBJ and SBJ samples. Combining the results of qualitative and quantitative analysis, it showed that the sulfur fumigation has a significant effect on BJ.
Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; Fumigation ; Least-Squares Analysis ; Principal Component Analysis ; Sulfur

BACKGROUND/OBJECTIVES: Heavy alcohol consumption causes the development of alcoholic liver disease (ALD), a neglected but important public health problem. Many studies have pointed out that probiotics could improve gut health, which is also considered to be a cause of ALD. Therefore, this study screened the probiotics, Lactobacillus casei GKC1 (GKC1), L. fermentum GKF3 (GKF3), Bifidobacterium lactis GKK2 (GKK2), L. rhamnosus GKLC1 (GKLC1), L. paracasei GKS6 (GKS6), and L. plantarum GKM3 (GKM3), for their potential benefits in alleviating ALD for applications to disease prevention. SUBJECTS/METHODS: C57BL/6N mice were divided into 8 groups (n = 6 in each): normal control, positive control (alcohol-diet fed), and treatments of feeding probiotics GKC1, GKF3, GKK2, GKLC1, GKS6, and GKM3 under an oral dose 0.82 g/kg B.W. per day by oral gavage. The experiment was conducted for 8 weeks, and the concentrations of alanine aminotransferase (ALT), aspartate aminotransferase, triglyceride (TG), and total cholesterol (TC) in mice were measured. The glutathione (GSH), catalase (CAT), and histology were analyzed after sacrifice. RESULTS: The results showed a decrease in the serum ALT, liver TG, and liver TC levels in the GKS6, GKM3, and GKLC1 groups compared to the positive control. In addition, the decreasing GSH and CAT levels were inhibited in the GKS6 and GKM3 groups. The histopathological results showed that all probiotics could reduce the accumulation of liver fat. Furthermore, there was a significant difference in GKLC1 with lower stomach damage compared to the alcohol-fed mice without any addition of probiotics. CONCLUSIONS GKLC1, GKS6, and GKM3 can be used as supplements for alleviating the development of ALD.

Objective To construct a specific small interfering double-stranded RNA(siRNA) expression vector of Caspase-12 and to evaluate inhibitory effect of this siRNA on caspase-12 mRNA activity.Methods Three groups of siRNA targeting different gene sites of caspase-12 were designed and synthesized chemically.Mouse hepatoma cell line,Hepa1-6,was transfected with the siRNA by 24 h.Reverse transcription-polymerase chain reaction(RT-PCR)was performed to analyze the inhibi- tion of caspase-12.Then the most effective siRNA was selected and the two template sequences for the siRNA were inserted into pRNAT-H1.1Neo expression vector.The recombinant plasmid, referred to as pRNAT-casp12,was verified by PCR analysis and sequencing.The expression of caspase-12 at mRNA and protein level,after transfection with pRNAT-casp12 by 48 h and 72 h respectively,were analyzed by using real-time PCR and Western blotting.Results The chemically synthesized siRNA*1 and siRNA*3 could inhibit mouse hepatoma cell caspase-12 mRNA by 59.9% and 39.6%(P

OBJECTIVEThe aim of this study was to evaluate the correlation of protein expressions of CXC chemokine receptor 4 (CXCR4), vascular endothelial growth factor-C (VEGF-C) and cytokeratin 19 (CK-19) with lymph node metastasis (LNM) in patients with hepatocellular carcinoma (HCC), and their survival.

METHODSThe expressions of CXCR4, VEGF-C and CK-19 in HCC patients with (n = 123) or without (n = 145) LNM were determined using tissue microarray and immunohistochemical staining. The relationship between clinicopathological features and CXCR4, VEGF-C and CK-19 were analyzed. Evaluation of immunostaining was performed semiquantitatively by visual assessment.

RESULTSThe UICC T stage, and expressions of nuclear CXCR4, VEGF-C and CK-19 were independent risk factors for LNM. Nuclear CXCR4, VEGF-C and CK-19 expression were predictive factors for LNM in HCC patients. In patients with LNM, the median survival time was 15.1 months for patients with high nuclear CXCR4 expression and 24.5 months for those with low nuclear CXCR4 expression. The median survival time was 15.1 months for patients with high tumor VEGF-C expression and 31.1 months for those with low tumor VEGF-C expression. The median survival time was 12.0 months for patients with positive CK-19 expression and 19.2 months for patients with negative CK-19 expression. Patients with high nuclear CXCR4, VEGF-C or CK-19 expression had significantly poorer prognosis than those with low expression (all P < 0.05). PVT, UICC T stage and expressions of nuclear CXCR4, VEGF-C, and CK-19 were independent prognostic factors.

CONCLUSIONIncreased protein expressions of nuclear CXCR4, VEGF-C, and CK-19 are independent risk factors for developing lymph node metastasis, and they are significantly correlated with LNM and poor outcome in HCC patients.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Nucleus ; metabolism ; Female ; Follow-Up Studies ; Humans ; Keratin-19 ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Receptors, CXCR4 ; metabolism ; Risk Factors ; Survival Rate ; Vascular Endothelial Growth Factor C ; metabolism