OBJECTIVE: To investigate the effects of Shenshuning Recipe (SSNR) on gene expression of hepatocyte growth factor (HGF) in renal tissues in rats with glomerulosclerosis. METHODS: Glomerulosclerosis was induced in 42 rats by unilateral nephrectomy and intravenous injection of doxorubicin. Then these 42 rats were randomly divided into three groups: untreated group, SSNR-treated group and benazepril-treated group. Another eight rats were included into sham-operation group. The rats in the SSNR-treated group and the benazepril-treated group were fed SSNR or benazepril respectively for 8 weeks. The levels of 24 h urine protein (Upr), serum creatinine (Cr) and blood urea nitrogen (BUN) of rats in each group were examined. The renal morphological changes were observed under microscope, and the diameter of glomerular capillary, mesangial matrix and glomerulosclerosis index were analyzed by image analysis software. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the gene expression of HGF in the renal tissues. RESULTS: The levels of 24 h Upr, serum Cr and BUN in the untreated group were remarkably increased than those in the sham-operation group (P<0.01). The pathological morphological changes in the untreated group showed that the glomerulosclerosis was diffused around the renal tissue and the capillaries were shrunk. The expression level of mesangial matrix was up-regulated and the glomerulosclerosis index was 3.32+/-0.35. The expression level of HGF mRNA in the untreated group was obviously lower than that in the sham-operation group (P<0.05). The levels of 24 h Upr, serum Cr and BUN in the SSNR-treated group and the benazepril-treated group were remarkably decreased as compared with those in the untreated group, while the expression levels of HGF mRNA were both obviously higher than that in the untreated group (P<0.01). The pathological morphological changes in the SSNR-treated group and the benazepril-treated group were both alleviated. There was no significant difference in therapeutic effect between the SSNR-treated group and the benazepril-treated group. CONCLUSION: Shenshuning Recipe can up-regulate the expression of HGF mRNA, decrease the mesangial matrix, and improve the renal function, so that it may retard the development of glomerulosclerosis.

Objective To investigate the effects of preoperative selective gastric arterial chemoembolization on the tumor vasculature in patients with gastric carcinoma.Methods 40 patients with gastric cancer were divided into two groups :(1)gastric arterial influsion(GAI)group;and (2)gastric arterial chemoembolization(GAE)group.The peripheral vein plasma tumor necrosis factor(TNF ?) and thrombomodulin(TM)were determined before radiologic intervention(RI),and1d,3d after R1;and the gastric vein plasma TNF ? and TM were also determined during operation.The curative surgical resection of gastric cancer was performed between 7 and 10 days after treatment.Stomach histological alterations were observed postoperatively.Results Compared with GAI group ,the plasma TNF ? levels increased markedly at all the time points,and gastric vein plasma TM levels decreased siginificantly in GAE group (P

OBJECTIVE: (1)To obtain the perfusion parameters of hepatocellular carcinomas(HCCs), peritumour livers and normal livers by multi-slice CT(MSCT)and to investigate their characteristics and clinical significances;(2)To investigate the correlation among perfusion parameters, survivin expression, microvessel density(MVD)and pathologic grade of HCCs. METHODS: A total of 31 patients with HCC (5 well-differentiated HCCs, 17 moderately differentiated HCCs, and 9 poorly differentiated HCCs) and 10 normal liver were studied. All underwent CT plain scan, perfusion scan, and conventional enhancement scan of the whole liver using 16-slice spiral CT (Philips Brilliance 16). Perfusion parameters were obtained by time-density curves (TDC) of region of interest (ROI) through the perfusion scans. Tissue sections of HCCs and their corresponding peritumour liver tissues of the 31 patients were detected by immunohistochemistry (SABC methods) for protein expression of survivin and MVD, and 10 normal liver tissue sections were as used as negative controls. The correlation among the perfusion parameters, survivin expression, MVD and pathology grade were analysed. RESULTS: (1)The mean values of HAP, HPP, TLP, and HAI of HCCs were 27.50 mL/(min.100 mL), 19.37 mL/(min.100 mL), 46.87 mL/(min.100 mL), and 60.38%, respectively. The mean values of those of the peritumour livers were 14.93 mL/(min.100 mL), 55.70 mL/(min.100 mL), 69.63 mL/(min.100 mL), and 21.51%, respectively. The mean value of those of the normal livers were 12.22 mL/(min.100mL), 74.56 mL/(min.100 mL), 86.78 mL/(min.100 mL), and 14.00%, respectively. The values of HAP and HAI of HCCs were significantly higher than those of the peritumor livers and the normal livers(P<0.01), and the HPP and TLP of HCCs were significant lower than those of the normal livers(P<0.01).The increase of HAP and decrease of HPP of peritumor livers were both significant compared with that of the normal livers(P<0.05). The HAP, HPP, and HAI of HCCs were significantly different from those of peritumor livers (P<0.01)except TLP. (2) Survivin expression in HCCs was detected in 23/31(74.1%), which was significantly higher than that in corresponding non-cancerous adjacent liver tissues and normal liver tissues (P<0.01). Survivin expression was positively correlated with MVD in HCCs. (3) HAP values were significantly and positively correlated with survivin expression (r=0.932,P<0.01)in HCCs.(4)The values of HAP and HAI were correlated with the pathologic grade in HCCs, and those values were increased gradually(P<0.05) among well differentiated HCCs, moderately differentiated, and poorly differentiated HCCs. CONCLUSION CTPI can quantitatively reflect abnormal blood supply of HCCs, which will be helpful for the detection and differentiation of lesions. CT perfusion parameters well correlate with survivin expression, MVD, and the pathologic grade in HCCs, which illustrate that CTPI could hopefully be used to evaluate the angiogenesis and biological behaviors of HCCs prospectively, noninvasively, and dynamically.
Adult ; Aged ; Carcinoma, Hepatocellular ; blood supply ; diagnostic imaging ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; biosynthesis ; genetics ; Liver Neoplasms ; blood supply ; diagnostic imaging ; metabolism ; pathology ; Male ; Microvessels ; diagnostic imaging ; Middle Aged ; Perfusion Imaging ; methods ; Survivin ; Tomography, Spiral Computed

Objective:To investigate the regulatory effects of ring finger protein 43 (RNF43) on CD8 + T cell-mediated anti-tumor immune reaction in melanoma. Methods:RNF43 gene was over-expressed and knockdown in mouse melanoma cells line B16-OVA by lentivirus infection; In vivo proliferation of mouse melanoma cells line B16-OVA in the Lv-Ctrl-OE, Lv-RNF43-OE, Lv-Ctrl-KD and Lv-RNF43-KD groups was detected by subcutaneous tumorigenesis assay in mice, and the expression levels of CD8 + T cells perforin and interferon γ (IFN-γ) in tumor immune microenvironment of melanoma were detected by flow cytometry; The expression levels of β-catenin and programmed death-ligand 1 (PD-L1) mRNA in cells were detected by quantitative real-time PCR assay; The effect of RNF43 on the transcriptional regulation of PD-L1 was detected by dual-luciferase reporter gene assay. Results:Stable RNF43 over-expressing and RNF43 knockdown mouse melanoma cells lines Lv-RNF43-OE and Lv-RNF43-KD were successfully constructed. The results of subcutaneous tumorigenesis experiment in mice showed that the tumor mass of the Lv-RNF43-OE group was (0.08±0.06) g, which was significantly smaller than that of the Lv-Ctrl-OE group [ (1.04±0.52) g], with a statistically significant difference ( t=3.71, P=0.032) ; The tumor mass of Lv-RNF43-KD group was (1.94±0.29) g, with no statistically significant difference ( t=-1.70, P=0.164) compared with that of the Lv-Ctrl-KD group (1.15±0.74) g. The flow cytometry results showed that the fluorescence intensity of CD8 + T cell perforin in the Lv-RNF43-OE group was 9 034 ± 2 628, which was significantly higher than that in the Lv-Ctrl-OE group (3 847 ±1 637), with a statistically significant difference ( t=-3.35, P=0.015) ; The fluorescence intensity of CD8 + T cell perforin in the Lv-RNF43-KD group was 966±247, which was significantly lower than that in the Lv-Ctrl-KD group (2 226±646), with a statistically significant difference ( t=3.16, P=0.034) ; The fluorescence intensity of IFN-γ of CD8 + T cell in the Lv-RNF43-OE group was 2 422±429, which was significantly higher than that of 1 688±324 in the Lv-Ctrl-OE group, with a statistically significant difference ( t=-2.73, P=0.034) ; The fluorescence intensity of IFN-γ of CD8 + T cell in the Lv-RNF43-KD group was 614 (454, 863), with a statistically significant difference ( Z=-1.96, P=0.050) compared with 1 159 (1 152, 2 068) in the Lv-Ctrl-KD group. The results of quantitative real-time PCR showed that the relative expression level of β-catenin mRNA in the Lv-RNF43-OE group was 0.67±0.16, which was significantly lower than that of 1.00±0.11 in the Lv-Ctrl-OE group, with a statistically significant difference ( t=2.98, P=0.041) ; The relative expression level of PD-L1 mRNA in the Lv-RNF43-OE group was 0.32±0.09, which was significantly lower than that of 1.00±0.09 in the Lv-Ctrl-OE group, with a statistically significant difference ( t=9.13, P=0.001). The results of the dual-luciferase reporter gene assay showed that the PD-L1 promoter luciferase activity in the pCMV6-NC, RNF43, RNF43+β-catenin and β-catenin groups were 1.00±0.00, 0.84±0.00, 1.49±0.00 and 1.57±0.03 ( F=2 218.33, P<0.001). Further pairwise comparison showed that compared with the pCMV6-NC group, PD-L1 promoter luciferase activity was significantly lower in the RNF43 group ( P<0.001) and significantly higher in the RNF43+β-catenin and β-catenin groups ( P<0.001; P=0.003) ; compared with the RNF43 group, PD-L1 promoter luciferase activity was significantly higher in the RNF43+β-catenin group ( P<0.001) . Conclusion:RNF43 may reduce the expression of PD-L1 mRNA in melanoma by inhibiting the expression of β-catenin and promote CD8 + T cell-mediated anti-tumor immune reaction.

OBJECTIVE：To investigate t he effects of different compatibility ratio of Gardenia jasminoides to fermented soybean on the content of genistein and total flavonoids ，and to investigate the compatibility regularity of Zhizichi decoction. METHODS：The decoction method was used to prepare the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4，m/m，the same hereinafter ）. UPLC-MS/MS method was used to determine the content of genistein in Zhizichi decoction with different compatibility ratio and corresponding fermented soybean single decoction. UV method was used to determine the content of total flavonoids in Zhizichi decoction with different compatibility ratio and corresponding gardenia single decoction and fermented soybean single decoction. RESULTS ：The established method had good linearity ， precision，repeatability，stability and accuracy. Compared with single decoction ，the content of genistein in the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4）was decreased to different extents ， while the content of total flavonoids was increased to different extents. With the increase of fermented soybean ，the content of genistein in the decoction increased at first and then decreased. When the compatibility ratios of G. jasminoides to fermented soybean were 1 ∶ 1 and 1 ∶ 2，the content of genistein in the decoction was the highest （all 0.071 μg/mL）. With the increase of fermented soybean ，the content of total flavonoids in the decoction did not change regularly ；when the ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of total flavonoids in the decoction was the highest （1.861 μg/mL）. CONCLUSIONS ： When the compatibility ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of flavonoids in the decoction is the highest.

ObjectiveTo investigate the effect of total alkaloids of Corydalis saxicola on a rat model of lipopolysaccharide（LPS）-induced depression， as well as the pharmacokinetic characteristics of 8 of its major components. MethodTwenty-four male SD rats were randomly divided into normal group， model group， fluoxetine group（10 mg·kg^-1） and total alkaloids of C. saxicola group（210 mg·kg^-1）， with 6 rats in each group. In addition to the normal group， the rats were injected intraperitoneally with LPS to establish the inflammation model of depression， and the drug administration was started 1 week after modeling， and the administration groups were gavaged according to the corresponding dose， and the normal and model groups were intragastric administration with equal volume of distilled water， and the administration was performed along with the modeling. After two weeks of continuous administration， the effect of total alkaloids of C. saxicola on the behavior of depressed rats were tested by sucrose preference， forced swimming and open field experiments， the levels of tumor necrosis factor-α（TNF-α）， interleukin（IL）-1β and IL-6 in serum of rats were determined by enzyme-linked immunosorbent assay（ELISA）， the histopathological changes of rat hippocampus were observed by hematoxylin-eosin（HE） staining. After the last administration， blood was collected from orbit according to the set time， and ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS） was established to simultaneously detect the concentrations of dehydrocavidine， tetrahydropalmatine， coptisine， palmatine， jatrorrhizine， berberine， berberrubine and epiberberine in plasma， and drug-time curves were drawn. The pharmacokinetic parameters were analyzed by DAS 2.0 software. ResultCompared with the normal group， the model group exhibited a decrease in sucrose preference rate， total distance traveled in the open field， as well as an increase in swimming immobility time and serum inflammatory factor expression（P<0.01）. In contrast， compared with the model group， rats in each administration group showed an increase in sucrose preference rate and total distance traveled in the open field， a decrease in swimming immobility time， and a reduction in serum inflammatory factor expression（P<0.05， P<0.01）. Additionally， HE staining results revealed that neurons in the hippocampus of rats from the model group were characterized by loss， disorganization and residual vacuoles， whereas those from the total alkaloids of C.saxicola group displayed an increase in number with orderly arrangement and clear cytoplasm. Pharmacokinetic results showed that the time to peak（t_max） and half-life（t_1/2） of the 8 active ingredients were 0.19-2.06 h and 3.71-8.70 h after continuous administration of total alkaloids of C. saxicola. Among them， the area under the curve（AUC_0-∞） of tetrahydropalmatine was the highest and the t_1/2 was the shortest， and the AUC_0-∞ of coptisine， palmatine， jatrorrhizine， berberine， berberrubine and epiberberine were low. The curves of dehydrocavidine， coptisine， palmatine， berberine and epiberberine showed obvious double peak phenomenon. ConclusionTotal alkaloids of C. saxicola can improve the depression-like behavior of rats， inhibit the expression of inflammatory factors in serum， improve the pathological injury of hippocampus， and has the antidepressant effect. Meanwhile， the effective site is absorbed quickly and eliminated slowly in the depressed model rats， and the efficacy is maintained for a long time.