Objective The method of data mining and sorting analysis was used to analyze and summarize the drug experience of Professor Li Zhenhua in the treatment of chronic atrophic gastritis (CAG). Methods Professor Li’s medical records of effective diagnosis and treatment of 139 CAG patients were collected. Find-Replace method of Microsoft office word 2007 was used to count the major syndromes and main prescription of CAG. SPSS statistical software was adopted to perform entry-analysis-descriptive statistics and data analysis of and main syndrome, main formula and frequency of administration so as to obtain the commonly used drugs, commonly used prescription and drug laws of CAG. Results Professor Li Zhenhua believed that the clinical syndromes of CAG included the disharmony of liver-stomach-spleen syndrome, the damp heat of spleen-stomach syndrome, the deficiency and damp heat of spleen syndrome, the liver depression and spleen deficiency syndrome, the deficiency of spleen-stomach syndrome, the liver and stomach yin deficiency and qi stagnation syndrome, the stagnant heat of liver-stomach syndrome and the blood stasis of stomach meridian syndrome;the commonly used drugs were:bupleurum, white peony root, orange peel, licorice, poria, skullcap, ginger, fried atractylodes, golden thread, prepared pinellia, licorice, lily, stir-baking Sanxian, nutgrass galingale rhizome, heterophylly falsesatarwort root, combined spicebush root, Chinese date, tangshen, immature orange fruit, prepared rhizome pinellize without adjuvant, and oyster shell..The commonly used prescriptions were: Xiaochaihu decoction, Sini powder, Chaihu-Guizhi-Longgu-Muli decoction, Chaihu-Shugan powder, Huanglian-Wendan decoction, Banxia-Xiexin decoction, Xiaoyao powder, Xiangsha-Liujunzi decoction. Conclusion Professor Li pay attention to treat spleen and stomach disease from liver by clearing heat and removing dampness from spleen and stomach. He used the dialectical methods like invigorating qi and strengthening the spleen, regulating qi digestion, activating blood flow to eliminating blood stasis.

E.coli single-stranded DNA-binding protein (SSB) plays an important role in replication, recombination and repair of DNA and is thus crucial for the survival of the bacteria.We described a high expression and efficient purification scheme and kinetic assay of interaction with its substrate, single-stranded DNA (ssDNA). A ssb gene (537 bp) for encoding SSB was obtained by PCR amplification from E.coli K-12 genome. The expression vector of the fusion protein SSB was constructed by attaching ssb gene to pQE30. SSB fusion protein was expressed in M15 E.coli strain induced by IPTG. SDS-PAGE analysis revealed that the expected protein with a molecular weight 20.6kDa was soluble and amounted to about 30% of the total bacterial protein. SSB protein was purified by immobilized metal (Ni2+) chelation affinity chromatography and the purity was about 90%. The resulting SSB protein was a correctly folded tetramer analyzed by gel filtration. It could bind ssDNA with equilibrium dissociation constant (KD) of 4.79×10-7 mol/L as determined by surface plasmon resonance.

To investigate the effect of the basic knowledge of chemistry teaching for preparatory international students,bilingual teaching based on Chinese-English languages and multimedia teaching,such flexible applications are carried out,which increases the students' listening,speaking,reading and writing skills in Chinese and helps them to adapt to the academic environment in China,and to master basic chemistry knowledge efficiently so that students can lay a good foundation for undergraduate program.

OBJECTIVE To study the effect of nano-alumina(nano-Al2 O3 )on mitophagy in primary cortical neuronal cells from Wistar newborn rats. METHODS The purity of neuronal cells was detected by immunohistochemistry,and the lactate dehydrogenase(LDH)assay was performed to determine the viability of the cells treated with 13 nm nano-Al2 O3 0.5 mmol·L-1 for 12,24 and 48 h,respectively. The mitochondrial membrane potential(MMP)was detected by flow cytometry analysis . The ultrastructure of mitochondria and mitophagy vacuoles was observed by transmission electron microscopy(TEM). Auto-phagic vacuoles were observed by dansylpentanediamine(MDC)staining and the expression of autoph-agy related protein Beclin1 and LC3Ⅱ/ Ⅰ was determined by Western blotting. Mitophagy was observed by Lysotracker and Mitotracker staining respectively. RESULTS More than 95% cells were neuronal cells. The activity of LDH in the supernatant liquid exposed to nano-Al2 O3 for 12 and 24 h groups was sig-nificantly increased compared with the control group(P﹤0.05). After exposure to nano-Al2 O3 ,the mito-chondrial membrane potential was significantly decreased compared with the control group( P ﹤0.01). The results of TEM displayed mitochondrial swelling and the formation of vacuoles and mitophagy in nano-Al2 O3 groups. MDC positive fluorescence particles were observed and the expression of autophagy related protein Beclin1 and LC3Ⅱ/ Ⅰ was increased in nano-Al2 O3 groups compared with the control group( P ﹤ 0. 05 ). The result of Lysotracker and Mitotracker colocalization showed the fusion of mitochondria and lysosomals. CONCULSION Nano-Al2 O3 may induce autophagy and mitochondria damage in neuronal cells while the damaged mitochondria may be removed by mitophagy.

Objective To investigate the clinical efficacy of purging fu-organs traditional Chinese medicine (TCM)therapy for treatment of patients with severe pneumonia and sthenia-heat. Methods According to random number table method,71 patients with sthenia-heat of severe pneumonia were divided into a treatment group (35 cases)and a control group(36 cases). Conventional basic treatment was given to both groups,and additionally, small chengqi decoction was applied nasogastrically for the therapy in treatment group for 2 weeks. The clinical pulmonary infection score(CPIS),Marshall score,integration score of TCM syndromes and the mortalities in 28 days and 60 days were used to compare the clinical efficacy of the two groups. Results With the prolongation of treatment,the CPIS,Marshall score and integration score of syndromes in the two groups were gradually decreased. In treatment group,CPIS and Marshall scores were lower than those of control group on the 4th day ,and there were statistically significant differences(CPIS score:5.8±1.7 vs. 6.8±1.9,Marshall score:5.3±2.3 vs. 6.6±2.7,both P<0.05);the above 2 scores in treatment group were also lower than those of control group on the 7th and 14th day after treatment(7th day CPIS score:5.3±1.5 vs. 5.6±1.4,Marshall score:5.1±1.9 vs. 5.7±1.8;14th day CPIS score:3.9±1.7 vs. 4.4±2.3,Marshall score:4.2±1.9 vs. 4.9±2.5),but there were no statistically significant differences(all P>0.05). In addition,the integration scores of syndromes were significantly decreased on the 4th, 7th and 14th day in the treatment group significantly lower than those in the control group(4th day:7.6±2.3 vs. 10.6±2.7,7th day:7.4±2.5 vs. 9.2±2.1,14th day:6.1±1.9 vs. 8.3±2.4,all P<0.05). However,there were no statistically significant differences in mortality rates in 28 days and 60 days respectively between control group and treatment group(28 days:16.7% vs. 11.4%,60 days:25.0% vs. 20.3%,both P>0.05). Conclusion Purging fu-organs therapy not only can decrease the CPIS and Marshall scores of patients with sthenia-heat of severe pneumonia,but also can improve their syndromes.

OBJECTIVE To explore the potential neurotoxicity of nano-alu mina (<50 n m)in vivo, we treated the ICR mouse with the nano-alu mina to investigate the mitochondrial da mage of nerve cells on morphology and function.METHODS Adult male mice were exposed to nano-alu mina (<50 n m)of 0,25,50 and 75 mg·kg -1 by nasal instillation for 1 month.Then we observed the mitochondrial ultra-structure of the nerve cells in CA3 region of hippoca mpus,and measured the mean dia meter in every group.The activities of Na +-K +-ATPase and Ca2 +-Mg2 +-ATPase were tested by the determination of the inorganic phosphorus,which was the deco mposition product of ATPase.Western blot analysis was used to detect the expression of COX-Ⅳ,Beclin1 ,LC3Ιand LC3Ⅱ.RESULTS Co mpared with 0 and 25 mg·kg -1 groups exposed to Al2 O3 nanopartilces (Al2 O3 NPs),the mitochondria of CA3 region in hip-poca mpus in 50 mg·kg -1 group beca me ede matous and swollen with sparse and broken cristae sur-rounding the nuclear,and the mean dia meter was higher(0.49 ±0.02 μm,P <0.05).But co mpared with 50 mg·kg -1 group,the mitochondria in 75 mg·kg -1 group beca me s maller with inner cristae of high density,and the mean dia meter was lower(0.36 ±0.02 μm,P<0.05).The enzy me activity of the mito-chondria in cerebral cortex decreased dose-dependently with exposure,the activities of Na +-K +-ATPase in 50 and 75 mg·kg -1 groups(6.37 ±0.22 kU·g -1 protein,5.48 ±1 .53 kU·g -1 protein)and Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups (3.21 ±0.99 kU·g -1 protein,3.28 ±0.15 kU·g -1 protein)were lower than the 0 mg·kg -1 group(P<0.05).Meanwhile,the Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups showed lower activities in co mparison with the 25 mg·kg -1 group.The 75 mg·kg -1 group expressed higher level of the COX-Ⅳ protein 1 .35 ±0.66(P<0.05)than other groups.Both expression of Beclin1 protein and rate of LC3Ⅱ/LC3Ⅰin 75 mg·kg -1 group were more than the 0 mg·kg -1 group. CONCLUSION The mitochondrial dysfunction may be the potential neurotoxicity of nano-alu mina,and the da maged mitochondria were cleared by autophagy.

Objective:To clone human anti-ricin antibodies from large phage antibody library.Methods:Panning for a large phage library against ricin toxin was conducted to select specific antibodies against ricin. The binding activities and specificities were tested by ELISA method. Soluble ScFvs were prepared through infecting E coli. HB2151 with the selected phage antibodies and induction with IPTG. Results:Forty positive clones were obtained after 5 rounds of panning, and 12 clones had specific binding ability to ricin toxin. DNA fingerprinting showed 7 different band patterns indicating 7 different positive clones. DNA sequencing showed that variable regions of these ScFvs belonged to different subgroups.Conclusion:Human anti-ricin antibodies were successfully obtained from large phage antibody library.

Objective To study the effect of processing time on the content of five active ingredients in vine-gar frankincense.Methods Acetic acid,3－acetyl－β－lactic acid,3－acetyl－α－lactic acid,11－carbonyl－β－lactic acid 5 kinds of active ingredients in vinegar frankincense were determined.Results The results showed that four kinds of lactic acid (α－boswellic acid,β－boswellic acid,3－acetyl－β－lactic acid and 3－acetyl－α－lactic acid)showed an increasing trend (from 16.88mg/g to 23.05mg/g,40.35mg/g to 61.05mg/g,11.02mg/g to 18.17mg/g,19.78mg/g to 25.08mg/g),11－carbonyl－β－lactic acid showed a decreasing trend (6.98mg/g to 5.86mg/g),with the increasing of processing time (5,10,15,20 and 30min).Conclusion 30 min was the best time to prepare the balsamic vinegar frankincense.

Minocycline, a tetracycline antibiotic, is now known to protect cells via an anti-inflammatory mechanism. We further explored this effect using an in vitro model of ischemia-like injury to neurons. Coculturing neurons with microglia, the brain's resident immune cell, modestly increased cell death due to oxygen and glucose deprivation (OGD), compared to neurons alone. Treatment of cocultures with minocycline decreased cell death to a level significantly lower than that of neurons alone. Treatment of cocultures with minocycline or inhibitors of various immune mediators, also led to decreased cell death. Importantly, treatment of neuron cultures without added microglia with these same inhibitors of tissue plasminogen activator, matrix metalloproteinases, TNF-alpha and inducible nitric oxide synthase as well as minocycline also led to decreased cell death. Thus, anti-inflammatory treatments appear to be directly protective of neurons from in vitro ischemia.
Cell Death ; Coculture Techniques ; Glucose ; Ischemia ; Matrix Metalloproteinases ; Microglia ; Minocycline ; Neurons ; Nitric Oxide Synthase Type II ; Oxygen ; Tetracycline ; Tissue Plasminogen Activator ; Tumor Necrosis Factor-alpha

OBJECTIVETo study the effect of arsenic on neuronal cell apoptosis and the mRNA and protein expression of calpain 1, calpain 2, and cyclin-dependent kinases 5 (cdk5)/p25 and to provide a scientific basis for the research on neurotoxic mechanism of arsenic trioxide (As2O3).

METHODSPrimary cultured rat neurons were divided into untreated control group, dimethyl sulfoxide (DMSO) solvent control group, and 1, 5, and 10 µmol/L As2O3 treated groups. Eight hours after being treated with As2O3, cell apoptosis rate was determined by flow cytometry, the mRNA expression of calpain 1, calpain 2, cdk5, and p35 was measured by real-time fluorescence quantitative PCR, and the protein expression of calpain 1, calpain 2, cdk5, p35, and p25 was measured by Western blot.

RESULTSCompared with those in the untreated control group and DMSO solvent control group, the cell apoptosis rates in the 5 and 10 µmol/L As2O3 treated groups were significantly increased (P < 0.05). The mRNA expression levels of calpain 1 were 6.36±3.26, 7.11±5.13, and 7.47±2.59 in the 1, 5, and 10 µmol/L As2O3 treated groups, respectively, and the mRNA expression levels of cdk5 were 1.27±0.19, 1.54±0.04, and 1.79±0.21 in the 1, 5, and 10 µmol/L As2O3 treated groups, respectively, which were significantly higher than those in the untreated group (0.72±0.15 and 1.77±0.87) and those in the DMSO solvent control group (0.96±1.23 and 1.18±0.09) (P < 0.05). The mRNA expression levels of p35 in the 1 and 5 µmol/L As2O3 treated groups were 2.17±0.59 and 2.51±0.51, respectively, which were significantly higher than that in the untreated control group (1.26±0.37) (P < 0.05). The protein expression levels of calpain 1 were 0.37±0.10, 0.42±0.13, and 0.51±0.18 in the 1, 5, and 10 µmol/L As2O3 treated groups, respectively, which were significantly higher than those in the untreated control group (0.11±0.08) and DMSO solvent control group (0.13±0.07) (P < 0.05). In the 5 and 10 µmol/L As2O3 treated groups, the protein expression levels of cdk5 were 0.34±0.12 and 0.37±0.21, while the protein expression levels of p25 were 0.31±0.23 and 0.55±0.16, all of which were significantly higher than those in the untreated control group and DMSO solvent control group (P < 0.05). The protein expression levels of p35 were reduced in the 5 µmol/L As2O3 treated group (0.31±0.23) and 10 µmol/L As2O3 treated group (0.26±0.16), as compared with those in the untreated control group and DMSO solvent control group (P < 0.05). The mRNA and protein expression of calpain 2 showed no significant differences between all groups (P > 0.05).

CONCLUSIONThe calpain 1-cdk5/p25 pathway may be involved in the process of As2O3-induced neuronal cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenic Poisoning ; Arsenicals ; Calpain ; metabolism ; Cells, Cultured ; Cyclin-Dependent Kinase 5 ; metabolism ; Neurons ; drug effects ; metabolism ; Oxides ; toxicity ; Rats