Effects of carbamazepine on BDNF expression in trigeminal ganglia and serum in rats with trigeminal neuralgia / 中南大学学报(医学版)

Objective:Trigeminal neuralgia(TN)is a severe chronic neuropathic pain that mainly affects the distribution area of the trigeminal nerve with limited treating efficacy.There are numerous treatments for TN,but currently the main clinical approach is to suppress pain by carbamazepine(CBZ).Brain-derived neurotrophic factor(BDNF)is closely related to chronic pain.This study aims to determine the effects of CBZ treatment on BDNF expression in both the trigeminal ganglion(TG)and serum of TN via a chronic constriction injury of the infraorbital nerve(ION-CCI)rat model. Methods:The ION-CCI models were established in male Sprague-Dawley rats and were randomly divided into a sham group,a TN group,a TN+low-dose CBZ treatment group(TN+20 mg/kg CBZ group),a TN+medium-dose CBZ treatment group(TN+40 mg/kg CBZ group),and a TN+high-dose CBZ treatment group(TN+80 mg/kg CBZ group).The mechanical pain threshold in each group of rats was measured regularly before and after surgery.The expressions of BDNF and tyrosine kinase receptor B(TrkB)mRNA in TGs of rats in different groups were determined by real-time PCR,and the expression of BDNF protein on neurons in TGs was observed by immunofluorescence.Western Blotting was used to detect the protein expression of BDNF,TrkB,extracellular regulated protein kinases(ERK),and phospho-extracellular regulated protein kinases(p-ERK)in TGs of rats in different groups.The expression of BDNF in the serum of rats in different groups was detected by enzyme-linked immunosorbent assay(ELISA). Results:The results of mechanical pain sensitivity showed that there was no significant difference in the mechanical pain threshold in the right facial sensory area of the experimental rats in each group before surgery(all P>0.05).From the 3rd day after operation,the mechanical pain threshold of rats in the TN group was significantly lower than that in the sham group(all P<0.01),and the mechanical pain threshold of rats in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 CBZ mg/kg group was higher than that in the TN group(all P<0.05).The BDNF and TrkB mRNA and protein expressions in TGs of rats in the TN group were higher than those in the sham group(all P<0.05),and those in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than the TN group(all P<0.05).The p-ERK levels in TG of rats in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were significantly decreased compared with the TN group(all P<0.05).The BDNF and neuron-specific nuclear protein(NeuN)were mainly co-expressed in neuron of TGs in the TN group and they were significantly higher than those in the sham group(all P<0.05).The co-labeled expressions of BDNF and NeuN in TGs of the TN+ 80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than those in the TN group(all P<0.05).The results of ELISA showed that the level of BDNF in the serum of the TN group was significantly higher than that in the sham group(P<0.05).The levels of BDNF in the TN+80 mg/kg CBZ group,the TN+40 mg/kg CBZ group,and the TN+20 mg/kg CBZ group were lower than those in the TN group(all P<0.05).Spearman correlation analysis showed that the BDNF level in serum was negatively correlated with mechanical pain threshold(r=-0.650,P<0.01). Conclusion:CBZ treatment can inhibit the expression of BDNF and TrkB in the TGs of TN rats,reduce the level of BDNF in serum of TN rats and the phosphorylation of ERK signaling pathway,so as to inhibit TN.The serum level of BDNF can be considered as an indicator for the diagnosis and prognosis of TN.

Cloning of transcription factor PcFBA-1 in Pogostemon cabin and its interaction with FPPS promoter / 中国中药杂志

Farnesyl diphosphate synthase(FPPS) is a key enzyme at the branch point of the sesquiterpene biosynthetic pathway, but there are no reports on the transcriptional regulation of FPPS promoter in Pogostemon cabin. In the early stage of this study, we obtained the binding protein PcFBA-1 of FPPS gene promoter in P. cabin. In order to explore the possible mechanism of PcFBA-1 involved in the regulation of patchouli alcohol biosynthesis, this study performed PCR-based cloning and sequencing analysis of PcFBA-1, analyzed the expression patterns of PcFBA-1 in different tissues by fluorescence quantitative PCR and its subcellular localization using the protoplast transformation system, detected the binding of PcFBA-1 protein to the FPPS promoter in vitro with the yeast one-hybrid system, and verified its transcriptional regulatory function by dual-luciferase reporter gene assay. The findings demonstrated that the cloned PcFBA-1 had an open reading frame(ORF) of 1 131 bp, encoding a protein of 376 amino acids, containing two conserved domains named F-box-like superfamily and FBA-1 superfamily, and belonging to the F-box family. Moreover, neither signal peptide nor transmembrane domain was contained, implying that it was an unstable hydrophilic protein. In addition, as revealed by fluorescence quantitative PCR results, PcFBA-1 had the highest expression in leaves, and there was no significant difference in expression in roots or stems. PcFBA-1 protein was proved mainly located in the cytoplasm. Furthermore, yeast one-hybrid screening and dual-luciferase reporter gene assay showed that PcFBA-1 was able to bind to FPPS promoter both in vitro and in vivo to enhance the activity of FPPS promoter. In summary, this study identifies a new transcription factor PcFBA-1 in P. cabin, which directly binds to the FPPS gene promoter to enhance the promoter activity. This had laid a foundation for the biosynthesis of patchouli alcohol and other active ingre-dients and provided a basis for metabolic engineering and genetic improvement of P. cabin.

SSR loci information analysis in transcriptome of Andrographis paniculata / 中国中药杂志

To study the SSR loci information and develop molecular markers, a total of 43 683 Unigenes in transcriptome of Andrographis paniculata were used to explore SSR. The distribution frequency of SSR and the basic characteristics of repeat motifs were analyzed using MicroSAtellite software, SSR primers were designed by Primer 3.0 software and then validated by PCR. Moreover, the gene function analysis of SSR Unigene was obtained by Blast. The results showed that 14 135 SSR loci were found in the transcriptome of A. paniculata, which distributed in 9 973 Unigenes with a distribution frequency of 32.36%. Di-nucleotide and Tri-nucleotide repeat were the main types, accounted for 75.54% of all SSRs. The repeat motifs of AT/AT and CCG/CGG were the predominant repeat types of Di-nucleotide and Tri-nucleotide, respectively. A total of 4 740 pairs of SSR primers with the potential to produce polymorphism were designed for maker development. Ten pairs of primers in 20 pairs of randomly picked primers produced fragments with expected molecular size. The gene function of Unigenes containing SSR were mostly related to the basic metabolism function of A. paniculata. The SSR markers in transcriptome of A. paniculata show rich type, strong specificity and high potential of polymorphism, which will benefit the candidate gene mining and marker-assisted breeding.

Comparison of development condition of different Liriope spicata seedlings / 中国中药杂志

The study identified the main morphological index of the seedlings classification including seedling age,the root width and number of newborn buds and coarse roots, according to the local agricultural production techniques and assessment of Liriope spicata's growth and development condition. After carrying on K cluster analysis of the morphological, we separated the seedlings into two levels. The first level (Ⅰ)： the new talent with the root width exceeding two point five millimeters, the new born buds exceeding three, and with the coarse root exceeding one. The second level (Ⅱ)： the old talent with the root width below one millimeters, the newborn buds below two and without coarse root. The study surveyed the plants' growth index dynamics, as well as the yield and quality of the tuberous root. The experimental results suggested that the growth condition of seedling Ⅰwas better, the yield of earthnut higher, the quality of earthnut more excellent. The study lied the foundation of L. spicata's grading standards and standardized production.

Preliminary study on effect of Rhodiolae Crenulatae Radix et Rhizoma cell wall-broken decoction pieces on intestinal flora of mice / 中国中药杂志

This study aims to analyze and compare the effect of cell wall-broken decoction pieces, conventional decoction pieces and conventional powder of Rhodiolae Crenulatae Radix et Rhizoma on the intestinal flora of normal mice. The conventional bacterial culture and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) were adopted for the mice after the oral administration for 14 days. According to the bacterial culture results, the 1/8 dose cell wall-broken decoction pieces group showed fewer Enterococcus and Escherichia coli bacillus but more Lactobacillus and Bifidobacterium than the conventional decoction pieces group and the traditional powder group (P <0.05). Meanwhile, on the basis of the PCR-DGGE results, the 1/8 dose cell wall-broken decoction pieces group revealed the highest Shannon-Wiener index (H) and species richness (S) among the seven groups, with extremely significant differences compared with the normal group (P <0.01), significant differences compared with the conventional decoction pieces group and the conventional powder group (P <0.05) and a high intra-group similarity. In conclusion, the long-term intake of 1/8 dose Rhodiolae Crenulatae Radix et Rhizoma cell wall-broken decoction pieces showed a certain effect in regulating intestinal tract by promoting the growth of Lactobacillus and Bifidobacterium. Furthermore, the intestinal flora community will become more stable.

Effects of Pb2+ stress on seed germination & seedling growth of Rabdosia rubescens / 中国中药杂志

The seeds of Rabdosia rubescens were as the materials to research the impacts of different lead (Pb2+) concentrations(0, 135, 270, 540, 1 080 mg x L(-1)) on seed germination and seedling growth. The results show that: Low concentration of lead had no obvious effect on early germination of the seed, the germination vigor and germination speed were lightly higher but not significantly differed at the level of Pb concentration 135 mg x L(-1) with control group; Mid-high concentration of Pb solution (270-1 080 mg x L(-1)) significantly inhibited the seed germination and seedling growth, which reduced the seed germination rate, germination vigor, germination index, embryo root length and shoot length, growth index with increasing of Pb concentrations. There was a inhibitory effect on embryo shoot length and root length at mid-high lead concentrations stress, and stronger inhibitory effect on root , which was more sensitive than shoot to Pb stress(P < 0.05). Pb bioaccumulation coefficient (BC) was 0.76-2.59, increased with concentration of Pb; Pb enrichment in seedling mainly caused the growth inhibition. The fitting model predictive analyses show, the critical concentration of Pb, which causes the germination rate and biomass fresh weight reducing 10%, is 195.18, 101.65 mg x L(-1).

A high throughput coupled with high performance liquid chromatography-tandem mass spectrometry method for determination of aflatoxin B1, B2, G1, G2 in 10 traditional Chinese medicines / 中国中药杂志

As the dilution procedure was applied, a simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of aflatoxin B1, B2, G1, and G2 was successfully by performed in a total 83 samples of 10 traditional Chinese medicines (TCMs), which were collected from 5 different hospital pharmacies and 5 different medical stores in Guangzhou city. Matrix effects of these 10 TCMs were ranged from 80.23% to 115.5% in low, intermediate and high concentration levels, indicating that the negative effect was overcome in this study. Meanwhile, the analysis method was proved to be stable and reliable during the whole analysis using Semen Armeniacae Amarum spiked 3 concentration levels of standard solution as quality control samples and the RSD < 6.6% was obtained. The contamination levels of 83 investigated samples were 13.89% and 17.02% in hospital pharmacies and medical stores, respectively. The result was presented to provide relevant reference and supplement to those researchers in TCMs analysis and screening.

Cross-validation of the osmotic pressure based on Pitzer model with air humidity osmometry at high concentration of ammonium sulfate solutions / 药学学报

The osmotic pressure of ammonium sulfate solutions has been measured by the well-established freezing point osmometry in dilute solutions and we recently reported air humidity osmometry in a much wider range of concentration. Air humidity osmometry cross-validated the theoretical calculations of osmotic pressure based on the Pitzer model at high concentrations by two one-sided test (TOST) of equivalence with multiple testing corrections, where no other experimental method could serve as a reference for comparison. Although more strict equivalence criteria were established between the measurements of freezing point osmometry and the calculations based on the Pitzer model at low concentration, air humidity osmometry is the only currently available osmometry applicable to high concentration, serves as an economic addition to standard osmometry.

Study of achieving multiple models of plasma treatment by remoulding B.BRAUN Diapact CRRT / 国际生物医学工程杂志

Objective To study how to use the machine of B.BRAUN's Diapact continuous renal replacement therapy (CRRT) to achieve multiple models of plasma treatment by analyzing the characteristics of the machine.Methods Based on the machine of B.BRAUN's Diapact CRRT,assisted by extra single pump,the pipeline connection of the machine was reformed and the appropriate parameter was reset.Results After the reconstruction,the new equipment could succeed in achieving special plasma treatment such as double filtration plasmpheresis (DFPP)and plasma adsorption (PA).Conclusions The reconstructed equipment can attain special plasma treatment.Compared with plasma exchange (PE),the equipment has some characteristics such as few syndromes and without external plasma.According to its safety and stability,this new method is suitable for clinical application.

Separation and molecular identification of fungal contamination on surface of 15 Chinese herbal medicines / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate fungal contamination on the surface of Chinese herbal medicines and explore an appropriate method for fast and efficient identification of contaminant fungi.</p><p><b>METHOD</b>Chinese herbal medicines were first washed and the washing solution was plated onto potato dextrose agar (PDA) to obtain the pure isolates. For molecular identification, two new pairs of specific primers were designed according to ITS region of fungi genome sequences. The strains were identified through polymerase chain reaction (PCR) and sequence analysis.</p><p><b>RESULT</b>Fifty fungal strains were obtained from the surface of 15 Chinese herbal medicines with the percent of contaminated samples of 93.3%. Twenty-seven strains among them were successfully identified.</p><p><b>CONCLUSION</b>Fungal contamination on the surface of Chinese herbal medicines is quite common. Although different fungal species were isolated, the genus Aspergillus was the predominant. The primer pairs developed in this study are compatible and can be used to identify fungal species from the surface of Chinese herbal medicines.</p>

Determination of aflatoxin B1, B2, G1, G2 in armeniacae semen amarum by high-performance liquid chromatography-tandem mass spectrometry / 中国中药杂志

A simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was established for simultaneous determination of aflatoxins (AFB1, AFB2, AFG1, AFG2) in Armeniacae Semen Amarum and the application was performance in 11 samples collected from different markets, medical stores and hospitals. The sample was extracted with 84% acetonitrile/water and 250 microL extraction was directly injected into a LC-MS/MS system without further purification procedure after being redissolved with methanol. The LC separation was performed on a C18 column with a linear gradient elution program of 4 mmol x L(-1) NH4 Ac-0.1% formic acid solution and menthol as the mobile phase. Selected reaction monitoring (SRM) was used for selective determination of the four aflatoxins on a triple quadruple mass spectrometer, which was operated in positive ionization modes. All the four aflatoxins showed a good linear relationship with r > 0.999 0, the average recoveries were between 87.88% and 102.9% and the matrix effect was ranged from 90.71% to 99.30% in low, intermediate and high levels. Furthermore, the higher recovery was obtained by the method reported in this study, comparing to the cleanup procedure with the Mycosep 226 purification column. Eleven samples collected were detected and the contamination levels of the AFB1 were between 1.590-2.340 microg x kg(-1) and the AF (B1 + B2 + G1 + G2) was ranged from 2.340 to 3.340 microg x kg(-1). In summary, the developed method was suitable to detect and screen AFB1, AFB2, AFG1, AFG2 in Armeniacae Semen Amarum.

Correlative factors of behavioral and psychological symptoms in patients with vascular dementia / 中华神经医学杂志

Objective To analyze the behavioral and psychological symptoms in patients with vascular dementia (VaD),investigate the relationship between behavioral and psychological symptoms and such correlative factors as cognitive disturbance,age,education level of the patients,and explore the internal relations among Neuropsychiatric Inventory (NPI) items. Methods NPI and Mini-Mental State Examination (MMSE) were introduced to assess the behavioral and psychological symptoms in 120patients with VaD and 61 healthy elders (control group). Results The VaD patient group had significantly higher scores in 8 items,including delusion,hallucination,agitation,dysphoria,apathy,irritability,aberrant motor and appetite/eating changes in the NPI list,as compared with the control group (P＜0.05),with the points ranked from the highest by dysphoria,apathy and irritability.Among the 8 items in the NPI list, delusion, apathy and aberrant motor were significantly correlated with cognitive disturbance; and aberrant motor was correlated with age and education level.Besides that,NPI factor analysis showed that 3 behavioral sub-syndromes,psychosis,mood disorder and behavioral disturbance,exited. Conclusion Behavioral and psychological symptoms were common in VaD patients,and the frequency and severity of appearing of behavioral and psychological symptoms were correlated with cognitive disturbance,age,and education level of different degrees; in addition,each sub-syndrome may have common pathophysiologic mechanism.

Influence of HBO therapy on level of ACA-IgG and thrombomodulin among parents with ischemic encephalopathy / 中华神经医学杂志

Objective To investigate the role of HBO on the change of levels of ACA and thrombomodulin among parents with ischemic encephalopathy. Methods Patients were divided into three groups: group A (patients with atherosclerosis and thrombosis),group B (patients with cardiogenic embolism) and group C (patients with small artery lesion). All patients were treated with HBO and regular drugs. And then sss scores were recorded and levels of ACA-IgG and thrombomodulin were detected by ELISA before and after therapy of HBO. Results The index of ACA-IgG and lelvel of TM in A,B and C group were higher than those in control group,there are differences among them(P＜0.05),and the level of them were higher in B group than in A and C group.There were significant differences in SSS score for brain function using different therapies in A、B and C group (P＜0.05), but there are no differences among them(P＞0.05).There were differences in the index of ACA-IgG and thrombomodulin using different therapies in A,B and C group(P＜0.05),however,there were no differences among them (P＞0.05). Conclusion HBO can alleviate clinical symptoms and improve recovery of neuron by decreasing the level of ACA-IgG and thrombomodulin in blood,but the effect is not significantly different for ischemic encephalopathy caused by different factors.

Predictive value of ABCD2 scale in cerebral infarction and death events after transient ischemic attack: a cohort medium-term follow-up study / 中华神经医学杂志

Objective To evaluate the predictive value of ABCD2 scoring system in cerebral infarction and death events in the medium-term after transient ischemic attack (TIA) of Chinese population. Methods One hundred and seventy-nine patients with TIA having complete clinical data,admitted to our hospital from January 2008, were chosen in our perspective study. The ABCD2 scale was applied to all the patients and used to observe the 18-month cerebral infarction risk events; according to these data, patients after TIA were divided into cerebral infarction group and non-cerebral infarction group; To the end of 18 months, according to the death event, the patients were divided into survival group and death group. The age, level of blood pressure, clinical features, duration of symptoms and history of diabetes were collected and compared between each 2 groups; the cutofflevel of ABCI2 scale for anticipating cerebral infarction risk and mortality was evaluated according to receiver operating characteristic (ROC) curve. Results Among the studied 179 patients, 52 patients appeared cerebral infarctions within 18 months of TIA; patients in the cerebral infarction group had significantly higher percentage of patients with blood pressure ≥ 140/90 mm Hg (86.5％), unilateral weakness (42.3％),duration of symptoms ≥60 minutes (55.8％), or duration of symptoms 10 to 59 minutes (40.4％), and diabetes (80.8％) as compared with patients in the non-cerebral infarction group (P＜0.05). According to the ROC curve, area under curve was 0.874 of ABCD2 scale for anticipating cerebral infarction risk, 95％confidence interval (CI) was 0.817-0.931 and the cutoff level of ABCD2 scale for anticipating medium-term cerebral infarction risk was 4.5. Among the studied 179 patients, 35 patients died within 18months of TIA; no significant difference in the scores of ABCD2 scale was noted between survival group and death group (P＞0.05); according to ROC curve, area under curve was 0.492 of ABCD2 scale for anticipating mortality, and 95％ CI was 0.389-0.596. Conclusion ABCD2 scale has predictive value in cerebral infarction event, while not in death event in the medium-term of TIA.

Expression of NgR in experimental allergic encephalomyelitis (EAE) and intervention of NgR (310)ecto-Fc in EAE / 中华神经医学杂志

Objective To investigate the role and the mechanism of NgR (310)ecto-Fc in experimental allergic encephalomyelitis (EAE). Methods EAE models were successfully induced in 90 Lewis rats and equally randomized into group A (without treatment), group B (giving NgR(310)ecto-Fc treatment 1 d after the success of model making) and group C (giving NgR(310)ecto-Fc treatment right after onset of the disease). The clinical scores, pathological changes of the animals were observed and compared before and after treatment. Changes of NgR expression and counts of NgR(310)ecto-Fc positive cells in the myeloid tissue were tested by immunohistochemistry before and after treatment.Results Clinical scores in group B (3.020±1.017, 1.365±0.127) and group C (2.853±0.958, 1.275±0.092) were significantly lower than those in group A (4.476± 1.525, 1.894+0.135) on the 15th and 25th d of success of model making (P＜0.05), while no significant differences on the clinical scores were noted between group B and group C. NgR expression was observed in the myeloid tissue of all groups; the counts of NgR(310)ecto-Fc positive cells in the myeloid tissue in group B (79.07± 10.31, 45.89±4.77) and group C (70.47±7.40, 40.63±4.15) were obviously decreased as compared with those in group A (101.12±11.97, 62.95±7.11) on the 15th and 25th d of success of model making (P＜0.05); while no significant differences on the counts of NgR (310)ecto-Fc positive cells were noted between group B and group C (P＞0.05). Conclusion NgR (310)ecto-Fc can alleviate the clinical symptoms of EAE by suppressing the expression of NgR, leading to no activation of myelin-related inhibitory factor (Nogo-A, MAG and OMgp), and inducing the growth of axons in EAE.