Optimization of Extraction Technology for Qinhao Nasal Drops by Multi-index Comprehensive Evaluation Method / 中国中医药信息杂志

Objective To optimize extraction process of Qinhao Nasal Drops by multi-index comprehensive evaluation Method. Methods With L9(34) orthogonal design, the content of rupestonic acid, chlorogenic acid, pseudoephedrine hydrochloride, total flavonoids and extract yield were set as indexes, and extraction technology of Qinhao Nasal Drops was optimized.Results Optimum extraction technology was:12 times amount of 60% ethanol, extracted three times. Conclusion The optimized extraction technology is simple, practical, and adapt to the production needs, which can be used as the basis for reasonable development of the preparation.

Combined Effects of Chronic Obstructive Pulmonary Disease and Depression on Spatial Memory in Old Rats / 中国医学科学杂志(英文版)

Objective To investigate the combined effects of chronic obstructive pulmonary disease (COPD) and depression on spatial memory in old rats, aiming to better understand the comorbidity of the two diseases in geriatric patients. Methods The SD rats were assigned into five groups: adult control group (n=6), elderly control group (n=6), elderly COPD group (n=6), elderly depression group (n=6) and elderly COPD with depression group (n=6). Smoking and chronic unpredictable mild stress (CUMS) with solitary support were used to induce COPD model, depression model, respectively, and the both were applied for the comorbidity model. Learning and memory deficits were assessed by Morris water maze (MWM) test. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum and hippocampus tissue were determined by Xanthinoxidase method and Thiobarbituric acid reaction (TBAR) method, respectively. Results The results of pulmonary histology, lung function, open-field test and sucrose consumption demonstrated the comorbidity models of COPD and depression in elderly rats were successfully established using smoking and CUMS with solitary support. Compared with the elderly control group, the group of COPD with depression had obviously longer time of latency and longer travel distance to reach the platform in MWM test (LSD-t=-10.116, P=0.000; LSD-t=-6.448, P=0.000). The SOD activity in serum and hippocampus decreased significantly (LSD-t=2.629, P=0.014; LSD-t=2.215, P=0.044) and the MDA content in serum and hippocampus increased significantly (LSD-t=-2.140, P=0.042; LSD-t=-2.070, P=0.049) in elderly COPD with depression group. Conclusions COPD in comorbidity of depression could induce spatial memory deficit in old rats. The mechanisms might be related to the overloaded and free radical metabolic imbalance. These results suggest a potential therapeutic target for comorbidity of COPD and depression in geriatric patients.

Molecular typing of Leptospira interrogans strains isolated from Rattus tanezumi in Guizhou Province, Southwest of China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To identify and type three leptospires isolated from Rattus tanezumi in Guizhou Province by using three molecular techniques (PFGE, MLVA, and MLST), reveal the molecular characteristic of causative agents of local leptospirosis and evaluate these three molecular methods based on their detection resolution and efficiency.</p><p><b>METHODS</b>Three Leptospira strains were isolated from the kidney of Rattus tanezumi and cultured with EMJH medium. PFGE, MLVA, and MLST assays were applied to type the three strains isolated from Rattus tanezumi in Guizhou Province.</p><p><b>RESULTS</b>PFGE, MLVA, and MLST typing showed that the three leptospiral isolates matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai. The findings of the genotyping methods were consistent. MLVA and MLST defined genotypes, whereas PFGE allowed the recognition of additional subgroups within the genotypes, and the findings of molecular typing were also consistent with those of traditional techniques.</p><p><b>CONCLUSION</b>Three leptospiral isolates from Guizhou Province matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai, and PFGE, MLVA, and MLST, as reliable molecular techniques for identifying and typing of Leptospira interrogans, would contribute to the active surveillance, outbreak investigation and source tracking for leptospirosis in Guizhou Province.</p>

Effects of icariin on Bcl-2 and Bax protein expressions and eosinophils apoptosis in bronchial asthmatic mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of icariin on Bcl-2 and Bax protein expressions and eosinophils apoptosis in bronchial asthmatic mice.</p><p><b>METHODS</b>48 female Balb/c mice were randomly divided into 6 groups, i.e., the normal control group, the model group, the Dexamethasone group, the low dose icariin group, the middle dose icariin group, and the high dose icariin group, 8 mice in each group. Bronchial asthma in mice were induced by intraperitoneal sensitization and challenged with nebulized ovalbumin (OVA). The mice of each treatment group were administrated with different doses of icariin by peritoneal injection from the first asthma sensitization (the 3rd week after the modeling) to the day before killing once every other day, while mice in the normal control group were administrated with physiological saline. The mice were killed after 6 weeks of treatment. The apoptosis of eosinophils and the Bcl-2 and Bax protein expressions of the lung tissues were detected by TUNEL and immunohistochemical assay respectively.</p><p><b>RESULTS</b>As compared with the model group, the apoptosis ratio of eosinophils were higher in the rest four treatment groups (P<0.05). The Bcl-2 protein positive areas in the lung tissues and the airway wall were significantly lowered (P<0.05). The Bax protein positive area significantly increased (P<0.05).</p><p><b>CONCLUSION</b>In bronchial asthmatic mice, icariin could enhance the apoptosis of eosinophils and lessen their infiltration by decreasing the expression of Bcl-2 protein and increasing the expression of Bax protein in lung.</p>

Establishment and application of TaqMan Real-time PCR for the detection of pathogenic Leptospira species / 中华流行病学杂志

Objective To develop and evaluate a TaqMan Real-time PCR method for the detection of pathogenic Leptospira species.Methods rrs gene of part fragment on 16S rRNA was used to design primers and TaqMan probe.The target gene was cloned into vector pMD19-T in order to make the standard curve and be used for quality control.To determine the specificity and specificity,DNA from Chinese Leptospira strains belonging to 15 pathogenic reference strains,21non-pathogenic reference strains,and 50 different serotypes of pathogenic isolates as well as 27 other micro-organisms were included in this study.Eight serial DNA dilutions from pathogenic Leptospira and DNA from 25 kidney tissues were detected by Real-time PCR and conventional PCR simultaneously.Results A Real-time PCR methodology was developed and optimised.All the pathogenic Leptospira gave a positive amplification.Non-pathogenic Leptospira and all the other micro-organisms were not amplified.The plasmid sensitivity of Real-time PCR and conventional PCR were 10 copy/μl and 104 copy/μl respectively.The DNA sensitivity of Real-time PCR and conventional PCR were 100 fg/μl and 1 ng/μl respectively.The kidney tissue detection of the two methods appeared to be exactly the same.Conclusion This research project successfully developed a Real-time PCR methodology with better sensitivity and specificity for the identification of pathogenic Leptospira,using the rrs gene.

Inflammatory airway features and hypothalamic-pituitary-adrenal axis function in asthmatic rats combined with chronic obstructive pulmonary disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Bronchial asthma (BA) and chronic obstructive pulmonary disease (COPD) are both inflammatory airway diseases with different characteristics. However, there are many patients who suffer from both BA and COPD. This study was to evaluate changes of inflammatory airway features and hypothalamic-pituitary-adrenal (HPA) axis function in asthmatic rats combined with COPD.</p><p><b>METHODS</b>Brown Norway (BN) rats were used to model the inflammatory airway diseases of BA, COPD and COPD + BA. These three models were compared and evaluated with respect to clinical symptoms, pulmonary histopathology, airway hyperresponsiveness (AHR), inflammatory cytokines and HPA axis function.</p><p><b>RESULTS</b>The inflammatory airway features and HPA axis function in rats in the COPD + BA model group were greatly influenced. Rats in this model group showed features of the inflammatory diseases BA and COPD. The expression of inflammatory cytokines in this model group might be up or downregulated when both disease processes are present. The levels of corticotrophin releasing hormone mRNA and corticosterone in this model group were both significantly decreased than those in the control group (P < 0.05).</p><p><b>CONCLUSIONS</b>BN rat can be used as an animal model of COPD + BA. By evaluating this animal model we found that the features of inflammation in rats in this model group seem to be exaggerated. The HPA axis functions in rats in this model group have been disturbed or impaired, which is prominent at the hypothalamic level.</p>

Lymphocyte immunological function and 24 T cell receptor V beta subfamilies expression in convalescent patients with severe acute respiratory syndrome / 中国组织工程研究

BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by a genetically novel coronavirus that is caused by acute infectious disease. It is not yet clear for the immunology function of SARS patients in their convalescent stage.OBJECTIVE: To study the effects on T lymphocyte, and the titer profiling of 24 T cell receptor (TCR) V β subfamilies expressions in SARS convalescent patients.DESIGN: A self-control observation.SETTING: Central Laboratory, Guangdong Provincial Hospital of Traditional Chinese Medicine.PARTICIPANTS: Seventy-six cured SARS patients who received treatment in the Second Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine between January and April 2003. All the patients corresponded to "clinical diagnostic criteria of atypical pneumonia", " diagnostic criteria of severe atypical pneumonia and discharge criteria" and "clinical diagnostic criteria and discharge criteria of severe acute respiratory syndrome". The involved patients, 30 male and 46 female, averaged (32±11 (years old. Another 10 subjects who simultaneously received health examination in the same hospital, 5 male and 5 female, aged (32±7(years, were involved in the study. Informed consents of detected items were obtained from all the subjects.METHODS：①Detecting the expression of 24 T cell receptor(TCR)V β subfamilies in SARS convalescent patients：Peripheral blood(2 mL) was collected from the healthy convalescent subjects,and EDTA-K2 was used as anticoagulant．In the flow cytometry delection tubes．10 μL various fluorescein-labeled mAb，such as anti-CD3,anti-CD4，anti-CD8,anti-CD25，anti-CD28，anti-HLA-DR，anti-CD3mAb conjugated with PC5，TCR Vβ1(PE+FITC)．Vβ2(PE+FITC)。Vβ3 (FITC)，Vβ4(PE+FITC)，Vβ5．1(PE+FITC),Vβ5．2(PE),Vβ5．3(PE)，Vβ7．1(PE+FITC)，Vβ7．2(FITC),Vβ8(FITC),Vβ9 (PE)，Vβ11(PE)，Vβ12(FITC)，Vβ13．1(PE),Vβ13．2(PE),Vβ13．6(PE+FITC)，Vβ14(FITC),Vβ16(FITC),Vβ17 (PE+FITC)，Vβ18(PE)，Vβ20(FITC)，Vβ21．3(FITC),Vβ22(PE+FITC)and Vβ23(PE)，was added in special flow tubes,and then 50 μL whole blood was added．The mixed solution was incubated away from light for 15 minutes．After erythrocytolysin being added，mixed solution was washed．Finally．cell deposit was dissolved in 300 μl phosphate buffer solution (PBS)．Coulter ESP flow cytometer was used for detection．For the analysis of TCR expression,an electronic gate was set on these cells and at least 5000 events per sample were collected．Three-color cytofluodmetric analysis was performed using a Coulter ESP flow cytometer．②Detecting the T cell subset,activated T and B cells,and the percentage of Ts and Tc cells：5000 cells were collected and used to calculate the expression of T cells (CD3,CD4 and CD8),the activated T and B cells(CD3+／CD25+,CD3+/HLA-DR+ and CD3-／HLA-DR+),as well as the percentage of Ts and Tc cells by Coulter ESP flow cytometer and its software．MAIN OUTCOME MEASURES：①The change of T cell subset(CD3，CD4，and CD8)from SARS convalescent patients．②The change of activated T and B cells(CD3+／CD25+，CD3+／HLA-DR+ and CD3-／HLA-DR+)．③The percentage of Ts and Tc cells(CD8+／CD28+，CD8+/CD28-)in convalescent patients．④Analysis of the 24 TCR V β subfamilies from SARS patients in convalescence．RESULTS：All data were explored to analyze the expression profiling of 24 TCR Vβ subfamilies,the data from 74 SARSpatients and 10 healthy controls were explored to other result analysis．①The detecting results of T celI subset：The percentage of CD4+T cell mean value was lower than the reference value[(33．33±6．64)％ vs．(43±9)％，P＜0．01]．The percentage of CD8+T cell mean value was higher than the refefence value[(34．07±6．40)％ vs．(30±9)％,P＜0．01]．② The expression of activated T and B cells：Percentage of HLA-DR+ T and B cell was Increased while the percentage of CD25+ T-cell was decreased compared with reference values．In 53 out of 74 patients,the percentage of CD25+ T cells was lower than the reference value,and 64 patients had a lower percentage in CD3+/CD25+ T cells．The percentages of CD3+/HLA-DR+ and CD3-／HLA-DR+ cells were higher than the normal reference value．T cells expressing higher CD3+／HLA-DR+ were found in 36 patients，and T cells expressing higher CD3-/HLA-DR+ were found in 30 patients．③The ratios of Ts and Tc cells：The percentage of Ts cells which expressed CD8+／CD28- was increased compared with reference value [(28．75±7．31)％ vs．(15．99±5．1)％，P＜0．01],while the percentage of Tc cells which expressed CD8+／CD28+ was decreased [(5．99±3．60)％ vs．(13．2±4．1)％，P＜0．01]．Thirty-nine patients were found to possess the lower Tc cells and forty-eight patients were found to possess the higher Ts cells．The ratios of both CD4+ and CD8+ T cells were in the normal reference value．④24 TCR Vβ subfamilies expressions in T cells：It was noteworthy that Vβ14 had a highest percentage in all 24 Subfamilies,and followed by Vβ 5．3,and Vβ 23 in the convalescent patients．The percentage of Vβ 14 was the highest in the normal controls,which was consistent with the results of SARS patients．But the other subfamilies expression patterns were different．There were significant differences between Vβ1,Vβ5．2,Vβ5．3,Vβ7．2，Vβ9，Vβ11,Vβ13．1,Vv13．2，Vβ17,Vβ18，Vβ22 and Vβ23．In the convalescent period，each TCR Vβ expression of SARS patients was higher than that of controls(P＜0．05-0．01)．CONCLUSION:In SARS convalescent patients,the increased CD8+CD28- T cell may elevate CD8+ T cell number;Meanwhile.the reduced CD3+ and CD4+ T cell number may be corresponding to the increased Ts cell number．For some inhibiting factor secreted by Ts cell was also increased．The usage pattern of 24 TCR Vβ subfamilies in SARS patients is different from that of control group．The increase of percentage of CD3+／HLA-DR+ and CD3-／HLA-DR+ T cell may be related to the late response of activated T and B cells.