In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P＜0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P＜0.01 or P＜0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P＜0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.

Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.

Objective To study the characteristics of dynamic susceptibility-contrast(DSC)MR perfusion curves,color images and perfusion values in pre-operative brain metastasis.Methods Twenty- eight brain metastases underwent DSC MR perfusion imaging by using a first-pass T_2~* echo-planar sequence. The patients' data were transferred to on-line workstation.Time-signal intensity curves,color perfusion maps and rCBV,rMTT values in both tumor parenchyma and peri-tumor edema were analyzed,and independent t- test was used and P0.05).Conclusion Different originated brain metastases have nearly same characteristics in DSC MR perfusion imaging.

Astragali Radix was firstly recorded in the "Shen Nong's Herbal Classic" as a top-grade and commonly used traditional Chinese medicine. Its frequently used slices include raw Astragali Radix and honey-processed products. In current studies, many reports were made on honey-processed Astragali Radix, whereas fewer study reports were made on the cutting process of Astragali Radix. Currently, because Astragali Radix is primarily cut by drug workers according to their operating experience, but with out specific cutting parameters, it is easy to cause the loss or mildew of active ingredients. As a result, the quality of Astragali Radix circulated in the market is not guaranteed, and the quality of their slices and preparations are hard to be controlled, which seriously impact the clinical efficacy. In response, this experiment was performed, in which the optimum cutting process of Astragali Radix was taken as the study objective, the Box-Benhnken central composite design in the response surface analysis was adopted, and the content and appearance character of astragaloside and calycosin-7-glucoside were regarded as the study indicators. Three factors, namely the softening time, the drying temperature and the drying time, were selected to optimize the cutting process of Astragali Radix and obtain the optimum cutting process parameters as follows: the softening time was 3 hours, the drying temperature was 50 degrees C, and the drying time was 4 hours. According to the verification test, the Astragali Radix cutting process is steady and feasible, which has certain significance for normalizing the cutting process of Astragali Radix.
Astragalus Plant ; chemistry ; Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Plant Roots ; chemistry

Background Nitrosamines (NAms) are highly carcinogenic and frequently detected in drinking water systems in China, indicating potential human health risk through drinking water. Objective To analyze the distribution of NAms in drinking water in Shanghai and to evaluate relevant human health risk. Methods A total of 94 samples of source water and 120 samples of finished water were collected in February (dry period) and August (wet period) of 2021 from 30 drinking water plants in Shanghai, and eight NAms were quantitatively analyzed by gas chromatography tandem mass spectrometry after solid phase extraction. Cancer risks for different age groups caused by NAms exposure through water were evaluated using Monte Carlo probabilistic method and carcinogens risk assessment model recommended by the United States Environmental Protection Agency. Results The concentrations of total NAms in source and finished water were 12.56-65.86 ng·L⁻¹ and 8.52-57.38 ng·L⁻¹, respectively. The average concentration of total NAms was higher in source water (33.50 ng·L⁻¹) than in finished water (22.07 ng·L⁻¹, P<0.05) during dry season, and lower in source water (16.90 ng·L⁻¹) than in finished water (21.02 ng·L⁻¹, P<0.05) during wet season. The average concentration of total NAms in source water during dry season was twice of that during wet season. The positive rate of N-nitrosodimethylamine (NDMA) was the highest (100%) among the eight NAms. The cancer risk of exposure to NAms in finished water in Shanghai was mainly from the ingestion route, which was mostly caused by NDMA. The cancer risk from exposure to NAms in water for children (median=4.32×10⁻⁵) was slightly higher than that for adults (median=3.34×10⁻⁵) and adolescents (median=2.27×10⁻⁵). The cancer risks of exposure to NAms in water for people of different ages were mainly (80% - 95%) at an acceptable level (1×10⁻⁶ - 1×10⁻⁴). Conclusion NAms contamination is positive in Shanghai’s drinking water and NDMA is the main contaminant. The removal of NAms in water by current water treatment process is season-dependent. The cancer risk of children exposed to NAms in water was slightly higher than that of adults and adolescents, but all at acceptable levels.

OBJECTIVETo localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16.

METHODSFive microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression.

RESULTSWith the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real-time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls.

CONCLUSIONResults reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE.

Adolescent ; Adult ; Alleles ; China ; Chromosomes, Human, Pair 16 ; genetics ; Cohort Studies ; DNA ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Microsatellite Repeats ; Middle Aged

OBJECTIVETo study the injection of NKG5SV gene to inhibit growth and metastasis of hepatocellular carcinoma (HCC).

METHODSNKG5SV gene was inserted into retroviral vector pLXSN by normal methods. LacZ gene was used as control. LCI-D20 tumor together with saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was subcutaneously inoculated to the nude mice. Tumor formation rate and tumor size were noted 35 days after inoculation. LCI-D20 tumor was inoculated subcutaneously. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was intratumorally injected respectively 10 days after inoculation. Tumor growth was observed 35 days after inoculation. Liver cancer was resected 22 days after intrahepatic inoculation. Saline, pLXSN-LacZ DNA or pLXSN-NKG5SV was respectively injected at incisal margin or intraspleen. Mice were killed 35 days after inoculation to observe tumor recurrence at incisal margin, intrahepatic metastasis and extrahepatic metastasis.

RESULTSTumor formation rate and tumor diameter(cm) were 1.76 +/- 0.11, 1.51 +/- 0.34, 0.33 +/- 0.04 in the control group, LacZ group, NKG5SV group respectively when tumor and different cDNA were inoculated together. Tumor diameter(cm) and weight(g) were 0.87 +/- 0.08, 0.83 +/- 0.05, 0.26 +/- 0.04; 0.43 +/- 0.06, 0.38 +/- 0.04, 0.08 +/- 0.06 in the control group, LacZ group, NKG5SV group respectively when different cDNA were injected into the LCI-D20 tumor. Sites with extrahepatic metastasis nidi, incisal margin recurrence tumor size(cm), intrahepatic metastasis nidi, metastasis involved hepatic lobes in the control group, LacZ group, NKG5SV group were 4.25 +/- 1.48, 4.25 +/- 1.04, 0.63 +/- 0.51; 1.51 +/- 0.27, 1.35 +/- 0.17, 0.81 +/- 0.17; 2.50 +/- 1.41, 2.38 +/- 1.06, 1.25 +/- 0.71; 2.13 +/- 0.99, 2.00 +/- 0.75, 1.38 +/- 0.74 respectively when NK cells were injected at incise margin. They were 4.38 +/- 1.85, 4.25 +/- 1.48, 1.00 +/- 0.75; 1.13 +/- 0.23, 0.97 +/- 0.29, 0.76 +/- 0.16; 2.50 +/- 1.41, 2.05 +/- 1.12, 0; 2.13 +/- 0.83, 1.75 +/- 0.88, 0 respectively when NK cell were injected intrasplenicly.

CONCLUSIONSNKG5SV gene can inhibit HCC growth and postoperative metastasis and recurrence.

Animals ; Antigens, Differentiation, T-Lymphocyte ; Cell Division ; drug effects ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; genetics ; Humans ; Injections ; Liver Neoplasms, Experimental ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; prevention & control ; Receptors, Immunologic ; genetics ; physiology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays

BACKGROUNDPrevious studies have suggested that interrupted clearance of nuclear DNA-protein complexes after cell death might initiate and propagate systemic lupus erythematosus (SLE). Deoxyribonuclease I (DNaseI) may be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover, thus preventing the onset of SLE. The purpose of this study was to genotype the single nucleotide polymorphisms (SNPs) of DNase1 and characterize its gene expression and alternatively spliced transcripts in Chinese patients with SLE in order to understand the pathogenic role of DNase1 in human SLE.

METHODSFour SNPs located at the 3' end of the DNase1 gene, as listed on the SNP website, were selected for analysis. Those SNPs with relatively high heterozygosity were chosen for genotyping in 312 Chinese SLE families using the Taqman minor groove binder (MGB) allelic discrimination method. Haplotypes were constructed and linkage disequilibrium tests were performed using GeneHunter. DNase1 mRNA expression was detected using real-time polymerase chain reaction (PCR), and alternatively spliced transcripts were isolated using capillary electrophoresis. Any effects the specific SNP haplotypes had on DNase1 gene expression and the alternatively spliced transcripts were also assessed.

RESULTSrs179982 and rs1053874 had high heterozygosity, about 0.5 in this Chinese cohort, while rs1059857 was also found to be heterozygous. Analysis of the haplotype combining rs179982-rs1030874 (C-G) and rs179982-rs1030874-rs1059857 (C-G-G) revealed a skewed transmission in favor of affected offspring. DNase1 gene expression was higher in SLE patients than in normal controls (P < 0.001), but this was not related to disease activity or SNP haplotype. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients differed from that of normal controls. Furthermore, different SNP haplotype combinations generated different transcript patterns in SLE patients.

CONCLUSIONSThe SNP haplotypes are in linkage disequilibrium in Chinese SLE patients and may induce the disease through a modification of DNase1 mRNA splicing rather than at the level of mRNA expression. There is a relatively unique transcript band in SLE patients independent of special haplotype, which suggests that other unknown factors might be involved in adjusting gene expression.

Adolescent ; Adult ; Alternative Splicing ; Deoxyribonuclease I ; genetics ; Female ; Haplotypes ; Humans ; Linkage Disequilibrium ; Lupus Erythematosus, Systemic ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Objective To investigate proton magnetic resonance spectroscopy(~1H-MRS)findings and value on dog's brain contusion and laceration.Methods Models of focal brain contusion and laceration in 10 dogs were established through hitting on the right frontal-parietal lobe with a freely drop of 200g weight at 1.3 m height.Serial examinations(1 h,24 h,72 h,5 day,8 day and 14 day after trauma)were performed with conventional MRI and ~1H-MRS.NAA/Cr,Cho/Cr and NAA/Cho rates were analyzed with GE system 1.5 T scanner and relative software.After examination,all dogs were executed to death. Pathological study was performed at local brain contusion.Results 1 h and 24 h-post trauma,NAA/Cr、Cho/Cr、NAA/Cho were significantly reduced(NAA/Cr 0.843?0.214,0.862?0.204,contralateral ones 1.069?0.284,1.048?0.232,t=-7.227,-6.718,Cho/Cr 1.181?0.224,1.243?0.134,contralateral 1.415?0.305,1.455?0.159,t=-4.332,-4.489,NAA/Cho 0.701?0.147,0.536?0.136, contralateral 0.832?0.245,0.613?0.165,t=-2.652,-2.665.P＜0.05).Microscopy showed focal petechial hemorrhage and necrosis,neuron loss,neuraxonal swelling and small glial cell slightly hyperplasia.Five day post trauma,Cho/Cr was significantly elevated in comparison with contralateral ones (1.517?0.197,contralateral 1.387?0.214,t=3.758.P＜0.05).Pathologically,inflammatory was obvious,peri-angiitis,granula tissue and fibrosis were seen.8—14 day later,NAA/Cr was not significantly reduced(0.895?0.105,0.875?0.153,contralateral 0.989?0.169,0.990?0.173,t=-2.909, -2.471.P＞0.05),Cho/Cr was significantly increased(1.457?0.168,1.572?0.374,contralaterl 1.334?0.174,1.366?0.352,t=7.312,3.201.P＜0.05)Inflammatory and gila1 hyperplasia was more significant,granuloma were seen.Lipid and Lac peak were not seen at all stages.Conclusion MRS could be a methods to monitor neuron injury and repair,and dynamically to detect the metabolic changes of brain contusion and laceration,reflecting injury severity and provide theory data for early treatment and predicting long-term outcome after trauma.

To establish quality standards of Euonymus fortunei, and supply scientific evidence for the quality control of Euonymus fortunei. Empirical and microscopic identification methods were adopted to observe morphological and histological characters. The contents of water, total ash, acid-insoluble ash and alcohol-soluble extractive were analysed according to the methods of Chinese Pharmaco- poeia (2010). Dulcitol and reference herbs were used to identify materia medica of Euonymus fortunei by TLC method. The total flavonol glycosides contents were analysed by HPLC method, using quercetin and kaempferol as reference substances. Quercetin and kaempferol were separated on a C18 column (4.6 mm x 250 mm, 5 μm) with methanol-0.1% formic acid(51:49) as the mobile phase and detected at 366 nm. The flavonoid aglycones content was then multiplied by a conversion coefficient, and the result was the total flavonol glycosides content. The macroscopical identification, microscopic features and TLC methods were proper. The average contents of water, total ash, acid-insoluble ash, alcohol-soluble extractive and total flavonol glycosides were 8.76%, 6.48%, 0.31%, 17.48% and 0.211% , respectively. The quality standards established on the basis of the research results were suitable for the quality evaluation of Euonymus fortunei.
China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Euonymus ; anatomy & histology ; chemistry ; Mass Spectrometry ; Quality Control