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OBJECTIVE To explore the material basis and potential mechanism of Kazakh classic prescription Wuzdekh （WZDK） in the treatment of enteritis. METHODS LC-MS/MS technology was used to analyze the chemical components in WZDK. Through network pharmacology and molecular docking technology， the main chemical components of WZDK were screened and the target was predicted； therapeutic effect and target of WZDK on acute enteritis were verified through in vivo experiments. The acute enteritis model of mice was induced by dextran sulfate sodium salt； the general condition of the mice was observed during administration and the disease activity index （DAI） score was calculated； pathological changes of the intestine and mRNA expression of core target were validated by HE staining and quantitative real-time PCR. RESULTS A total of 316 chemical components were obtained by LC-MS/MS. The core targets of network pharmacological analysis mainly included interleukin 1β（IL- 1β）， protein kinase B1 （AKT1）， tumor protein p53 （TP53）， IL-6， tumor necrosis factor （TNF） and so on. The results of molecular docking showed that chemical components such as mairin， lappadilactone， costunolide and dehydrocostus lactone were stable in binding to the core target. The results of in vivo experiment showed that， compared with model group， high dose （5.00 g/kg） of WZDK could significantly reduce the DAI score （P＜0.05）， improve inflammatory cell infiltration and mucosal tissue damage of colon tissue， and significantly down-regulated mRNA expressions of IL-6， TNF-α， IL-1β and TP53 in colon tissue（P＜ 0.05 or P＜0.01）. CONCLUSIONS Chemical components of WZDK such as mairin， lappadilactone， costunolide and dehydrocostus lactone may play the role of improving the imbalance of local inflammatory factors in the intestine and repairing damage of colonic mucosal tissue by down-regulating mRNA expressions of TNF-α， IL-6， IL-1β and TP53 in colon tissue.
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OBJECTIVE@#To construct a HIV-1 gp120 transgenic mice (gp120 Tg) with vimentin (VIM) gene knockout.@*METHODS@#Female HIV-1 gp120 Tg mice were mated to VIM heterozygote mice (F0). All the offspring mice were derived from these original founders so that both genotypes had the same mixed genetic background. The F1 mice were bred to generate of VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. PCR was performed for genotyping of the mice, and the expressions of VIM and gp120 in the brain tissues were examined using immunoblotting.@*RESULTS@#The results of PCR showed the presence of the target bands in VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. In VIM/gp120 Tg mice, gp120 expression was detected throughout the brain regions while no VIM expression was detected.@*CONCLUSIONS@#We generated gp120 transgenic mouse models with VIM gene knockout, which facilitate the exploration of the role of VIM in gp120-induced neurotoxicity.
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Animals , Female , Mice , Brain , Disease Models, Animal , HIV Envelope Protein gp120 , HIV-1 , Mice, Knockout , Mice, Transgenic , VimentinABSTRACT
Objective To investigate the role of HIV-1 envelope protein gp120 in cognitive im-pairment induced by neuronal damage. Methods Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neu-ronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohisto-chemical staining and behavioral analysis, respectively. Results In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuro-nal synaptic shortening and neuronal damage (P<0. 05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neu-rocognitive disorders. Conclusions HIV-1 gp120 might cause neuronal damage through activating the re-lease of inflammatory factor by microglia and involve in neurocognitive impairment.
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Objective@#To investigate the role of HIV-1 envelope protein gp120 in cognitive impairment induced by neuronal damage.@*Methods@#Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neuronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohistochemical staining and behavioral analysis, respectively.@*Results@#In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuronal synaptic shortening and neuronal damage (P<0.05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neurocognitive disorders.@*Conclusions@#HIV-1 gp120 might cause neuronal damage through activating the release of inflammatory factor by microglia and involve in neurocognitive impairment.
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OBJECTIVE:To establish a method for full-length cDNA library of Xinjiang Artemisia rupestris. METHODS:Mod-ified Trizol method was adopted to extract total RNA in young leaves of A. rupestris,it was transcribed into single-strand cDNA, and then synthesized into double-strand cDNA by long-distance polymerase chain reaction(LD-PCR)method. PCR product was di-gested by proteinase K and sfiⅠ,and then fractionated by CHROMA SPIN-400 columns. The cDNA longer than 0.4 kb were col-lected and ligated to phage λTriplE × 2,and then protein packaging was performed. Full-length cDNA library was established by SMART technology. 20 monoclonal were randomly selected from the library,and electrophoresis was used to determine the primary library titer,library capacity,recombinant positive rate and length of insert cDNA. RESULTS:The primary library titer was 1.94× 107 pfu/mL,library capacity was 0.97×107 pfu;recombinant positive rate of insert cDNA was 96% and length was 0.5-2.0 kb with an average of 0.9 kb. CONCLUSIONS:The established library is high in capacity and quality,which can provide basis for estab-lishing cDNA library of Xinjiang A. rupestris.
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OBJECTIVE:To establish a rapid,accurate and standardized DNA barcode identification method for the ferulic me-dicinal plants. METHODS:Genomic DNA was extracted from Ferula sinkiangensis K. M. Shen and Ferula fukanensis K. M. Shen,ITS2 sequences were amplified and sequenced,analytical similarity search method was conducted,13 species 26 samples gene ITS2 sequences of ferulic medicinal plants in GenBank database were downloaded and comparatively analyzed;interspecies and intraspecies genetic distances were calculated,and phylogenetic tree was developed for cluster analysis. RESULTS:According to calculation,species genetic distances of the 15 species ranged 0.009-0.230,average genetic distance was 0.018;results of clus-ter analysis showed DNA barcode ITS2 sequences can cluster the 32 kinds of ferulic medicinal plants into different classes. CON-CLUSIONS:The established DNA barcode ITS2 sequences identification method can accurately and rapidly identify the genuine and false samples of ferulic species.
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Objective To evaluate protective effects of hypothermic pulmonary protective solution with uli-nastatin on lung function during cardiopulmouary bypass (CPB) in the patients with congenital heart disease(CHD) and pulmonary hypertenion. Method Fifty-four children,who had CHD of left-to-fight shunts with moderate-se-rious pulmonary hypertension, were enrolled. They had been performed with the radical operation under CPB from September 2005 to December 2006 in the Department of Cardiovascular Surgery, Children' s Hospital of Zhejiang University. Moderate-serious pulmonary hypertension was defined as pulmonary-to-systolic pressure ratio > 0.45(Pp/Ps > 0.45). Fifty-four children were randomly divided into three groups. Patients in group A (n = 18)didn't receive pulmonary protective solution, and scrved as control; patients in group B (n = 18) were adminis-tered with pulmonary protective solution without ulinastatin;patients in group C (n = 18) were administered with pulmonary protective solution with ulinastatin. The serum concentrations of MDA and MPO were measured at five different time points:pre-operation, 0 h, 3 h, 6 h and 24 h in the intensive care unit (ICU) (T1～5). Patients'lung functions were monitored at T1 - T4. The time of mechanical ventilation was recorded. Results No one died in this study. The mean time of mechanical ventilation was shorter in the group B and group C than that in the group A. The MDA and MPO levels were lower in group B compared with group A at T4. The MDA level at T3-T5 and the MPO level at T4 was lower in group C than those in group A. There were no significant in MDA and MPO levels between group B and group C at five time point.A-aDO2 was lower in groups B and C than those in group A at T3 and T4, whereas at T4, A-aDO2 was lower in group C than that in group B. Cdyn was higher in group B at T3and group C at T3 - T4 than those in group A. Cdyn was lower in groups C than that in group B at T4.Condusions Lung perfusion with hypothermic protective solution during CPB can all lung injury and promote recovery after operation, especialy with ulinastatin.
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Objective To study the significance of c-met mRNA in axillary drainage after operations for breast cancer. Methods RT-PCR assay was used to examine c-met mRNA in axillary drainage after operations in 52 cases of breast cancer. The relationships between the expression of c-met and the tumor size, metastatic lymph nodes, the expressions of estrogen receptor (ER), progesterone receptor (PR) and c-erbB-2 were analyzed, respectively. In addition, the effect of douching operative field with 5-FU and distilled water on the expression of c-met mRNA was also analyzed. Results ①The proto-oncogene c-met mRNA could be detected in axillary drainage after operations for breast cancer by RT-PCR, and its positive rate was higher than that in routine pathological detection for micrometastasis in the axillary lymph nodes (P
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AIM: To establish the quality standard for Hanshi Tougu Tablets(Radix Aconiti Praeparata,Radix Aconiti Kusnezoffii Praeparata,Ramulus Cinnamomi,Rhizoma Atractylodis, Semen Sinapis,Radix Aucklandiae,Radix Glycyrrhizae). METHODS: Rhizoma Atractylodis,Semen Sinapis,Radix Glycyrrhizae in Hanshi Tougu Tablet were identified by TLC.The content of cinnamaldehyde and costunolide in Hanshi Tougu Tablets were determinded by HPLC. RESULTS: Rhizoma Atractylodis,Semen Sinapis,Radix Glycyrrhizae,Radix Aconiti Praeparata and Radix Aconiti Kusnezoffii Praeparata could be identified by TLC.Cinnamaldehyde showed a good linear relationship at the range of 2.104 ?g?mL~(-1)-52.60 ?g?mL~(-1),r=0.999 8(n=5), and the recovery was(99.62%,) RSD was(0.88%).Costunolide also showed a good linear relationship at a range of(0.036) mg?mL~(-1)-0.54 mg?mL~(-1),r=(0.999 9)(n=5),and the recovery was 96.10%,RSD was 0.15%. CONCLUSION: The method is simple,accurate and with strong specificity and can be used for the quality control of Hanshi Tougu Tablets.
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Objective: To investigate the clinical effect of Gynecologic Qianjin Tablets (Radix Codonopsis, Radix Angelicae sinensis, Folium Mahoniae, Caulis Millettiae Reliculatae, Herba Andrographis, etc.) for treating chronic pelvic inflammatory disease. Methods: 30 cases with chronic pelvic inflammatory disease in study group were treated by Gynecologic Qianjin Tablets, which has the effects of boosting Qi and blood and clearing away heat and wet in 2 weeks, 3 times per day and 6 tablets per time; and meanwhile, 30 cases treated by tablet of Jinji Tablets in control group were also observed. Results: The response rate in study group was 83.33% , while in control group was 63.33% ( P