Transport of Lipoamide and Lipoic Acid in Caco-2 Cell Monolayer Model / 中国药学杂志

OBJECTIVE: To study the transport of lipoamide (LAM) and lipoic acid (LA) in Caco-2 cell monolayer model in vitro. METHODS: Effects of LAM and LA on the survival rate of Caco-2 cells were investigated by MTS, the bi-directional transport of lipoamide and lipoic acid from the intestinal cavity side (apical side, AP) to the basal side (basolateral side,BL) was investigated. The cumulative transport volume, apparent permeability coefficient (Papp) and transport percentage were calculated,and the relationships between transport volume and concentration and time were further studied. RESULTS: The transport amounts of LAM and LA were increased in time-and concentration-dependent manners, the Papps of LAM and LA (AP→BL) were 2.443 44×10-5-2.392 91×10-5 and 8.179 78×10-6-7.897 25×10-6 cm•s-1, and the Papps(BL→AP) were 2.258 13×10-5-2.214 3×10-5 and 8.267 98×10-6-7.926 73×10-6 cm•s-1, respectively. CONCLUSION: In the transport test of Caco-2 cells, LAM is superior to LA, suggesting that it is well absorbed orally and has high bioavailability. But it is still necessary to verify the pharmacokinetic data in vivo.

Protective effect of paeoniflorin against PM2.5-induced damage in BEAS-2B cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of paeoniflorin against PM2.5-induced damage in BEAS-2B cells and explore the possible mechanism.</p><p><b>METHODS</b>With a factorial design, this study was performed to observe the protective effects of different doses of paeoniflorin against PM2.5-induced BEAS-2B cell growth inhibition and the effects of paeoniflorin on the contents of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS) in the cell cultures.</p><p><b>RESULTS</b>Exposure to increased PM2.5 concentrations caused significant decrease in the cell survival rate (P<0.05) with a clear dose-response relationship (r=-0.759, P<0.05). Treatment of the cells with paeoniflorin significantly attenuated PM2.5-induced inhibition of BEAS-2B cell survival (P<0.05), but the effect of paeoniflorin was not dose-dependent (P>0.05). PM2.5 exposure also significantly increased the contents of MDA and intracellular ROS (P<0.05), and paeoniflorin obviously antagonized these effects of PM2.5.</p><p><b>CONCLUSION</b>Paeoniflorin can protect BEAS-2B cells from PM2.5-induced growth inhibition, and the mechanism might be related to the anti-oxidant effects of paeoniflorin.</p>

Snitrosylating protein disulphide isomerase mediates increased expression of α synuclein caused by methamphetamine in mouse brain / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of Snitrosylation of protein disulphide isomerasec in methamphetamine (METH)-induced expression of alpha synuclein (αSN) in mouse hippocampus and striatum neurons.</p><p><b>METHODS</b>Forty C57BL/6 mice were randomized equally into saline control group, METH group, L-NNA (a NOS inhibitor) group and L-NNA plus METH group. All the agents were injected intraperitoneally at an interval of 12 h, and a total of 8 injections were administered; in L-NNA plus METH group, METH was injected 30 min after LNNA in each treatment. Western Blotting was used to detect the expression of nitric oxide synthase (NOS), αSN, PDI and Snitrosylation of protein disulphide isomerase (PDI-SNO) in the hippocampus and striatum of the mice, and nitric oxide (NO) levels were determined using a NO assay kit.</p><p><b>RESULTS</b>In METH group, the levels of NOS, PDISNO, αSN and NO all increased significantly compared with those in the control group (P<0.05). Combined treatment with L-NNA and METH, compared with METH alone, resulted in significantly lowered expression of NOS, NO, PDI-SNO and αSN in the hippocampus and striatum of the mice (all P<0.05). No significant differences were found in NOS, NO, PDI-SNO or αSN expressions among METH+L-NNA, L-NNA and control groups (P>0.05).</p><p><b>CONCLUSION</b>METH induces the activation of NOS and increases NO level to cause the occurrence of PDI-SNO, leading subsequently to increased expression of αSN in mouse striatum and hippocampus. L-NNA, the inhibitor of NOS, can partly relieve nervous system toxicity induced by METH.</p>

Effect of methamphetamine exposure on S-nitrosylation of protein disulphide isomerase in PC12 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of methamphetamine (METH) exposure on S-nitrosylation of protein disulphide isomerase and the neurotoxicity of METH in PC12 cells.</p><p><b>METHODS</b>PC12 cells were exposed to different concentrations of METH, and the cell viability was assessed using the cell-counting kit-8. PC12 cells exposed to METH in the presence of the NOS inhibitor N-nitro-L-arginine (L-NNA) were examined for cell viability and S-nitrosylation of protein disulphide isomerase using the biotin-switch method, and the changes in cell morphology were examined with HE staining.</p><p><b>RESULTS</b>METH exposure obviously decreased the cell viability and increased S-nitrosylation of protein disulphide isomerase, and the effect of METH was obviously inhibited by L-NNA treatment.</p><p><b>CONCLUSION</b>METH can cause obvious neurotoxicity and promote S-nitrosylation of protein disulphide isomerase in PC12 cells.</p>

Effect of low-selenium diet on expressions of CCR7, CD206 and CD163 in the liver and kidney of rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of low-selenium diet on the liver and kidneys of rats and explore the role of macrophage polarization into M1 and M2 phenotypes in liver and kidney injuries.</p><p><b>METHODS</b>Twenty-four rats (12 female and 12 male) were randomly divided into control group and low-selenium group and fed with normal chow (dietary selenium of 0.18 mg/kg) and low-selenium diet (dietary selenium of 0.02 mg/kg) for 109 days. After the feeding, the rats were sacrificed for HE staining to observe liver and kidney pathologies, and immunohistochemistry was performed for analyzing CCR7, CD206, CD163-positive cell numbers in the liver and kidneys.</p><p><b>RESULTS</b>The rats in low-selenium group showed severer fibrosis in the liver and kidney than the control group. In either male or female rats in low-selenium group, CCR7 and CD206 expressions in the liver were comparable with those in control group, but CD163 expression was lower than that in the control group (P<0.05 for both female and male rats). In the kidney, the proximal tubule showed a slightly higher while the distal tubule showed a slightly lower CCR7 expression in low selenium group than in the control group (P>0.05). In low-selenium group, a significantly lower CD163 expression in the distal tubule and a significantly higher CD206 expression in the proximal tubule were noted as compared with the control group (P<0.05 in both female and male rats). Compared with the control rats, the male rats in low-selenium group, but not the female rats, showed a significantly lower CD163 expression in the proximal tubule of the kidney (P<0.05); the female but not the male rats in low-selenium group show a higher CD206 expression in the distal tubule (P<0.05).</p><p><b>CONCLUSION</b>Low-selenium diet can cause liver and kidney fibrosis in rats and may inhibit macrophage activation into the M2 phenotype.</p>

A study on the single -tube multiplex RT -real time PCR assay in detecting rotavirus,astrovirus and hepatitis A virus / 预防医学

Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.

Rapidly detect and distinguish between norovirus G I and G II type with a pair of primers / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.</p><p><b>METHODS</b>A pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.</p><p><b>RESULTS</b>The results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.</p><p><b>CONCLUSION</b>The RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.</p>

A novel naphthalene derivative from Aloe barbadensis / 药学学报

To investigate the chemical constituents of A. barbadensis, aqueous extract of the plant was subjected to preparative medium pressure liquid chromatography (MPLC). The chemical structures were mainly determined by spectroscopic evidences (UV, IR, HR-MS, 1H NMR, 13C NMR, HSQC, 1H-1H COSY and HMBC) and chemical methods. A new O, O, O-triglucosylated naphthalene derivative, together with two known 6-phenyl-2-pyrone derivatives and four 5-methylchromones, were isolated and identified as 1-((3-((4- O-beta-D-glucopyranosyl)-beta-D-xylopyranosyloxymethyl)-1-hydroxy-8-alpha-L-rhamnopyranosyloxy)naphthalene-2-y])-ethanone (1), 10-O-beta-D-glucopyranosyl aloenin (2), aloenin B (3), aloesin (4), 8-C-glucosyl-(R)-aloesol (5), 8-C-glucosyl-7-O-methyl-(S)-aloesol (6), and isoaloeresin D (7). Compound 1 is a novel naphthalene derivative and named as aloveroside B, compounds 2-3 are isolated from this Aloe species for the first time.

Molecular characteristics of noroviruses causing outbreaks of acute gastroenteritis in Huzhou / 病毒学报

To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou. During April 2008 and February 2009, fecal specimens of patients collected from 2 outbreaks of acute gastroenteritis were tested for Norovirus by real-time RT-PCR. Partial sequence of RNA dependent RNA polymerase(RdRp) of the positive samples were amplified by RT-PCR, the PCR products were then purified, sequenced and phylogenetic analysis was conducted. Both genogroup II (GII) and genogroup I (GI) noroviruses were detected in 2 outbreaks. Phylogenetic analysis revealed that two of the GI norovirus strains isolated from 2008 belonged to genotype GI/2 and one of the GI Norovirus strain isolated from 2009 belonged to genotype GI/3. The other GIIú norovirus strains isolated from 2009 had high nucleotide identity with GIIb genotype that had been reported frequently in European countries during 2000 and 2001 and in Asian countries recently. These results suggested that the epidemic strains of norovirus isolated in Huzhou had a high degree of genetic diversity and prevalent genotypes at different times were also different. To our knowledge this is the first report of detecting GIIb variant in outbreaks of acute gastroenteritis in China.

Molecular characteristics of noroviruses causing outbreaks of acute gastroenteritis in Huzhou / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou.</p><p><b>METHODS</b>From 2008 to 2010, total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tested for Norovirus. Partial sequence of RNA dependent RNA polymerase (RdRp) of the positive samples were amplified by RT-PCR, then the PCR production were purified, sequenced and put into phylogenetic analysis.</p><p><b>RESULTS</b>50 of 119 specimens were positive for Norovirus by real-time RT-PCR. Out of those 50 Norovirus positive specimens, 9 were Norovirus Genogroup I (GI) positive, 35 were Norovirus Genogroup II (GII) positive, 6 was both Norovirus GI and GII positive. 12 PCR products for RdRp were selected for further studies on sequencing. Phylogenetic analysis revealed that the 5 GI norovirus isolates were belonged to genotype GI/2 and GI/3. Of the 7 GII norovirus isolates, 6 were belonged to genotype GII/4, 1 was belonged to genotype Glib.</p><p><b>CONCLUSION</b>Norovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.</p>

An investigation and analysis on an outbreak of acute gastroenteritis caused by genogroup I and II norovirus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the cause of an outbreak characterized by diarrhea and vomit in a middle school in Huzhou City.</p><p><b>METHODS</b>Comprehensive analysis was conducted based on field epidemiological study, clinical characteristics of the cases and laboratory test.</p><p><b>RESULTS</b>578 cases of acute gastroenteritis were found. The attack rate was 23.58%. The most frequently observed clinical symptoms were diarrhea, vomiting, abdominal pain and nausea. Some few had fever. Most cases had slight clinical symptom with a course from 1 to 3 days. The cases were distributed in every class, showing no phenomenon of clustering. Norovirus were detected in 11 out of 15 stool samples by using RT-PCR. 6 were genogroup II norovirus. 3 were genogroup I norovirus. enogroup I and II norovirus were detected at the same time in 2 stool samples (the same student with 2 tests). Case-control study showed that drinking unheated bottled water was risk factor (OR = 2.46, 95% CI = 1.19-5.23), and had a dose response relation with the disease (chi = 24.8 P < 0.01). The epidemic was controlled soon through isolating patients during treatment, providing boiled water, disinfecting and health education.</p><p><b>CONCLUSION</b>This was an infectious diarrhea outbreak caused by norovirus. The suspected transmission ways were drinking unheated bottled water and contact daily.</p>

Expression of rat protamine gene in MEL cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the expression of rat protamine (RP) gene in MEL cells and the effect on cell growth.</p><p><b>METHODS</b>Eukaryotic expression plasmid pCR-3.1-RP was constructed and transfected into MEL cells. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated.</p><p><b>RESULTS</b>Transfected MEL cells showed lower growth rate, mitotic index and colony-forming rate. Volumes of cells were reduced and reduction of RNA transcription was observed.</p><p><b>CONCLUSION</b>These results suggest that expression of RP in MEL cell may inhibit the cell growth and proliferation.</p>

Test and verify of piping cleaning effect in infusion solution productive process / 药学实践杂志

OBJECTIVE：To test and verify the piping cleaning effect in infusion solution productive process.METHODS:Targets of final washing water of pipe were determined.RESULTS:Targets of final washing water of pipe were that electrical conductivity of entrance of pipe was equal to that of exit of pipe essentially,limitation of bacteria and endotoxin were as follows:＜50CFU/ml,＜0.25EU/ml.CONCLUSIONS:According to the clean rules laid down by pharmaceutical Factory of Shanquan,Jinan,The desired cleaning effect of the pipe of infusion solution can be achieved.