Mechanism and experimental verification of Sijunzi Decoction in treatment of ulcerative colitis based on network pharmacology / 中国中药杂志

To explore the mechanism of Sijunzi Decoction in the treatment of ulcerative colitis(UC) based on network pharmacology. The active components and corresponding targets of Sijunzi Decoction were extracted with Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets were standardized with the help of Uniprot database. The related targets of UC were obtained through GeneCards database and Disgenet database, and the intersection targets of drugs and diseases were screened by R language. The visual regulation network of "active ingredient-disease target" of Sijunzi Decoction was constructed by Cytoscape software, and the protein-protein interaction network was constructed by STRING database. The functional enrichment analysis of gene ontology(GO) and the enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG) pathway were carried out on Bioconductor platform, and some of the targets were verified by animal experiments. Through database analysis, a total of 135 active components of Sijunzi Decoction, 114 predicted targets and 80 common targets with UC were obtained. The core target proteins included interleukin 6(IL-6), caspase-3(CASP3), vascular endothelial growth factor A(VEGFA), epidermal growth factor receptor(EGFR) and so on. GO functional enrichment analysis involved 102 items, which mainly affected transcription factor activity, enzyme activity, receptor activity and biochemical process regulation. KEGG pathway enrichment analysis showed that 120 items were involved in human cytomegalovirus infection, cancer, apoptosis, inflammation and other pathways. Mouse experiments showed that Sijunzi Decoction could down-regulate the expression of target proteins IL-6 and caspase-3 and inhibit intestinal epithelial cell apoptosis. The treatment of UC with Sijunzi Decoction is the result of the interaction among multi-components, multi-targets and multi-pathways. It is proved by experiments that Sijunzi Decoction may play an effective role by regulating the expression of IL-6 and caspase-3, and getting involved in apoptosis, inflammation and other pathways.

[The Effect of Chronic Intermittent Hypoxia on Cognitive Function and Prefrontal Cortex Neurons in Rats].

OBJECTIVE: To determine the effect of chronic intermittent hypoxia on cognitive function and prefrontal cortex neurons in rats. METHODS: 48 adult male Wistar rats were randomly divided into two groups: control group and 50 mL/L intermittent hypoxia group (50 mL/L CIH). Rats in the CIH group were placed in the low oxygen tank, simulating intermittent hypoxia environment. At 7 d, 14 d, 21 d, and 28 d, the learning and memory ability of the rats was assessed with the Morris water maze (MWM) test; the expressions of cysteinyl aspartate specific protease (caspase)-8 protein in their prefrontal cortex were determined using Western blot method; the apoptosis of neurons was detected by the TdT mediated UTP nick end labeling (TUNEL) method. RESULTS: Compared with the controls, the CIH rats had significantly prolonged escape latency at 14 d, 21 d, and 28 d (P<0. 05) and decreased target quadrant time (P<0. 05). The prolonged escape latency increased and target quadrant time shortened with length of exposure to hypoxia (P<0. 05). Compared with controls, the CIH rats had gradually increased caspase-8 in their frontal cortex neurons, peaked at 28 d (P<0. 05). The CIH rats showed obvious structural damage and reduced neuron density in their frontal cortex neurons. They had higher levels of nerve cell apoptosis (P<0. 05), with apoptosis index increasing with length of exposure to hypoxia (P<0. 05). CONCLUSION: Severe chronic intermittent hypoxia can lead to pathological changes of frontal,cortex of rats, possibly

Relationship Between the DPD and TS mRNA Expression and the Response to S-1-Based Chemotherapy and Prognosis in Patients with Advanced Gastric Cancer.

The aim was to determine changes in dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) mRNAs in the blood of advanced gastric cancer (AGC) patients to see whether these enzymes affected the patients' response to S-1-based chemotherapy and prognosis. For this purpose, pretreatment DPD/TS mRNA expressions were determined in 40 AGC patients using RT-PCR. The patients were then administered with S-1-based regimen (S-1 + cisplatin) and toxicities were recorded. The relationship between the DPD/TS mRNA expressions and the chemotherapy response, drug resistance, and prognosis was analyzed. The data show that DPD mRNA expression correlated significantly with Lauren type while TS mRNA expression correlated with distant metastasis. Patients with higher DPD and/or TS mRNA expression(s) showed poor response, while those with low DPD mRNA expression showed better response to the chemotherapy. Pooled analysis showed that the patients with low DPD/TS mRNA expressions had better therapeutic response. The incidence of bone marrow suppression, diarrhea, and oral mucositis was high in patients with low DPD mRNA expression. Median overall survival (OS) in 40 patients was 13.5 months. It was 17 months for low and 10 months for high DPD (P = 0.044) and TS mRNA expression (P = 0.047). Pooled analysis showed that the patients with both low DPD/TS mRNA expressions had longer OS (P = 0.001). In conclusion, the detection of DPD and/or TS mRNA expression can be used to predict the response to S-1-based chemotherapy, drug resistance, and prognosis in AGC patients as well as to help guide the individualized treatment of gastric cancer.

[Experimental study of the effect of adipose stromal vascular fraction cells with VEGF on the neovascularization of free fat transplantation].

OBJECTIVE: To investigate the effect of adipose stromal vascular fraction cells (SVFs) with VEGF on the neovascularization of free fat transplantation. METHODS: SVFs were obtained from subcutaneous fat and labelled with DiI. 0.3 ml autologous fat tissue was mixed with 0.2 ml cells: 1) autologous SVFs with VEGF (Group A); 2) autologous SVFs (Group B); 3) complete DMEM (Group C) And then the mixture was injected randomly under the back skin of 12 nude mice. The transplanted fat tissue in three groups was harvested at 2 months after implantation. Wet weight and diameter of fat grafts was measured. After HE and CD31 staining,blood vessel density, viable adipocytes and fibrous proliferation were observed. RESULTS: Trace of SVFs labeled by DiI in vivo could be detected by fluorescent microscope. The wet weight of fat grafts was (191.90 +/- 9.81) mg in group A, (177.01 +/- 10.50) mg in group B, and (92.05 +/- 8.30) mg in group C (P<0.01). The diameter of fat grafts was (0.49 +/- 0.24) cm in group A, (0.40 +/- 0.26) cm in group B, and (0.32 +/- 0.28) cm in group C (P<0.01). Histological analysis showed the blood vessel density was (14.58 +/- 2.06)/HPL in group A, (11.55 +/- 2.18)/HPL in group B, (7.87 +/- 1.55)/HPL in group C. Compared with group B and group C, group A had more adipose tissue with less fat necrosis and fibrosis and had significantly higher capillary density. CONCLUSIONS: The autologous adipose stromal vascular fraction cells with VEGF could improve the neovascularization of free fat significantly. It indicates a wide clinical application in the future.

[Investigation on an outbreak of measles caused by new virus (d11 genotype) imported from Myanmar].

OBJECTIVE: To study the relevant factors on an measles outbreak caused by imported new virus (d11 genotype) from Myanmar and to develop effective strategies and measures. METHODS: On-site investigation on the outbreak was carried out. RESULTS: There were four townships (66%) in Menglian county reported 15 cases of measles, with 7 cases aged 6 months to 5 years old, 2 cases with the history of measles vaccination (MV). Another 8 cases were 21 to 49 year-olds but their histories on immunization were unclear. 14 of the measles cases with Myanmar citizenship came to China for treatment. They were aged 10 months to 13 years old, with only one case had ever received MV vaccination. For all the 29 cases, except for one case who did not adopt the sample case of Myanmar, the remaining 28 patients were positive for measles IgM antibodies. 6 cases of measles virus RNA were detected in the amplified sequence which showed genotype d11, and was considered Myanmar imported wild virus. 184 people received the MV inoculation, with a rate of 61.96% and the serum samples showed a measles IgG antibody positive rate of 87.50%. Manner MV emergency vaccination was carried out timely in that county so the measles outbreak was effectively controlled. CONCLUSION: Imported measles cases from foreign countries might lead to epidemic, indicating the difficulty and challenge in the elimination of measles in our province. Emergent vaccination of MV could interrupt the transmission of the disease. Our experience showed that MV was effective in the prevention of d11 genotypes measles infection in the area.

[Status of enterovirus infection and molecular identification among healthy children at the areas bordering Myanmar, in Yunnan province, China].

OBJECTIVE: To explore the enterovirus infection status among healthy children under 15 years old in the border areas of Yunnan province that connecting Myanmar. METHODS: A total of 319 stool samples were collected from healthy children in the 10 entrance ports. Enterovirus was isolated from these stool samples and then poliovirus and adenovirus were serotyped by neutralization test using specific anti-sera. All the non-polio enteroviruses (NPEVs) were identified by partial sequencing of VP1 gene. RESULTS: All 53 enterovirus were isolated from 319 stool samples and 16.6% of them carried the virus. 23 polio virus (PVs) and 30 NPEVs were isolated with rates of carrying the virus were 7.2% and 9.4% respectively. 4 adenovirus were also isolated with a rate as 1.25%. 1 isolate could not be amplified by any Pan-enterovirus primers or by RT-PCR so was not able to be sequenced. The results of NPEVs sequencing showed that:1 isolate (3.3%) was classified into 1 serotype of HEV-A while 20 isolates (66.7%) were classified into 11 serotypes of HEV-B and 8 isolates (26.7%) were classified into 3 serotypes of HEV-C. However, we could not isolate any viruses that belong to HEV-D. nt. Result from the aa identify calculation showed that the nt and aa identification between isolates and corresponding standard strains were more than 75% and 85% respectively. The findings were similar to the international standards. CONCLUSION: Our results showed that the rate of carrying the enterovirus especially poliovirus in some areas of Yunnan province that bordering Myanmar was higher than that of rate through the routine acute flaccid paralysis detection system. Of the enterovirus isolated, HEV-B group appeared the predominant with the wide spread of enterovirus serotype. Some newer enterovirus were also detected such as EV73 (2 strains), EV75 (1 strain), EV80 (1 strain) and EV96 (4 strains).

[Molecular identification of human enterovirus 73 in Yunnan Province, the People's Republic of China].

Molecular typing was conducted according to the reported methods for 2 enteroviruses that were isolated from healthy children in the border areas of Yunnan Province with Myanmar. RT-PCR and sequencing were performed with 292/222 primers according to the Oberste's methods. The resulting sequences were blasted against the Genbank database and compared with all available enterovirus database. Analysis of homology at nucleotide and amino acid level identically suggested that the two enteroviruses are human enterovirus 73.

[Effect of different combinations of compound Danshen preparation on pharmacokinetic parameters of salvianolic acid B in rat plasma].

OBJECTIVE: A sensitive liquid chromatography-mass spectrometric (LC-MS) method for the quantification of salvianolic acid B in rat plasma was developed and validated. The effect of different combination on the salvianolic acid B were studied to investigate SSTG'pharmacokinetics mechanism. METHOD: Different combination methods were used for oral administration. The compounds were extracted by solvent in rat plasma and determined using LC-MS-MS method. The results were analysis by DAS software. RESULT: Chromatographic separation was achieved on a reversed-phase SB-C18 column with the mobile phase of acetonitrile-methanolformic acid (0.1%) and step gradient elution resulted. The analytes were detected by using an electrospray negative ionization mass spectrometry in the MRM mode. The method showed excellent linearity over the concentration range 0.5-1000 microg x L(-1) (R2=0.999). The low limit of quantification was 0.5 microg x L(-1). The extract recoveries of analytes were from 30.88% to 38.18%. The precisions, the accuracy and the stability all meet the requirements for all analytes. The HPLC-MS-MS technique provided an excellent method for the quantification of SAB in rat plasma. CONCLUSION: The method is simple, sensitive and rapid. The plasma concentration of Rg1 and Re were too low to determine. Ginseng & Yanhusuo and Ginseng & Danshen combination can decrease the AUC value and increase the Vd and CL.