AIM:To establish HPLC method for making a assay of nucleosines in Ningxinbao Capsules(Cepha-(lospovium) sinensis Chen.sp.nov). METHODS:The assay was performed on a C_(18) column(4.6 mm?250 mm,5 ?m).The mobile phase was composed of phosphate buffer(pH 7.8-8.0)-acetonitrile(83∶17).The flow rate was at 1.0 mL/min.The detection wavelength was at 260 nm. RESULTS:The linear ranges of uridine,adenosine and inosine lay in the concentration range of 4.168-83.36,4.480-89.60,5.230-104.6 ?g/mL,respectively(r=0.999 9). The average recoveries were 0.4%,1.8%,2.5%,respectively. CONCLUSION:The method is simple,rapid.It is suitable for the assay of nucleosines in Ningxinbao Capsules.

Objective To develop and validate the real-time fluorescent quantitative PCR(Q-PCR)method for the detection of 8 murine viruses. Methods The specificity,sensitivity and precision of the Q-PCR method were verified by four laboratories,and the virus simulated contamination test and blind sample detection were carried out simultaneously,of which the detection results were compared. The Q-PCR method was used to detect 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other products of murine origin. Results The Q-PCR method used for detecting 8 kinds of murine viruses had no cross reaction with the same family and genus or other murine viruses. Except the sensitivity of laboratory 2 to ectromelia virus(EctV/Mouse Pox,MPV)was 2 × 10~2copies/μL,the sensitivity of laboratory 2 to other 7 viruses and 3 other laboratories to 8 murine viruses was 2 × 10~1copies/μL. Except the inter-assay CV of the copy number of mouse adenovirus(MAdV)detected by laboratory 3 was 37. 58%,the intra-assay and inter-assay CVs of the Ct and copy number of other 7 viruses detected by laboratory 3 and those of 8 viruses detected by other 3 laboratories were all less than 25%.The sensitivity of virus simulated contamination test met the parameter requirements. The coincidence rate of blind sample detection results by 4 laboratories was 100%. All the 26 batches of monoclonal antibody cell lines for SARS-CoV-2 vaccine production and 15 batches of other murine derived products were negative for 8 murine viruses. Conclusion Q-PCR method for murine virus has good specificity,sensitivity and precision,and can be used for the detection of murine derived biological products.

BACKGROUND: Migraine is usually located in the course of Shaoyang channel, and the curative effect of treatment with methods of treating Shaoyang disease by dispersing and dispelling wind (DW) as well as relieving pain (RP) on migraine is prominent, but how about the effects of these two ways on thrombogenesis in vitro and the function of platelet aggregation (PA)?OBJECTIVE: To probe into the effect of treatment of Shaoyang disease and methods of DW and RP on thrombogenesis in vitro and function of PA throughthe experiments of thrombogenesis in vitro and function of PA in rats, and investigate the compatible significance of DS and DW as well as RP through formula compatible experiment.DESIGN: Completely randomized grouping design and controlled trial.SETTING: Department of Prescription, College of Basic Medical Sciences,Heilongjiang University of Chinese Medicine.MATERIALS: The experiment was conducted in the Laboratory of Department of Prescription, Heilongjiang University of Traditional Chinese Medicine from March to August 2000. Sixty healthy adult Wistar rats were randomly divided into 6 groups: Normal control group, Shaoyang treatment group (group Ⅰ ), DW and RP group (group Ⅱ ), high-dose and whole-prescription group (group Ⅲ), low-dose and whole-prescription group (group Ⅳ), and positive control group with 10 rats in each group.METHODS:① Positive control group:Gastric perfusion of 0.034 g/mL compound divaside slice suspension (made by Harbin Second Biochemical Pharmacy co., Ltd) was given to rats (0.39/kg); Group Ⅲ and Ⅳ (ingredients: Chaihu 20 g, huangqin 10 g, banxia 15 g, gancao 10 g, chuanqiong 20 g, tianma 15 g, xixin 5 g, quanxie 5 g, wugong 5 g): Gastric perfusion of 1.5, 0.75 g/mL of complete prescription was given to rats (17.40, 8.70 g/kg);Group Ⅰ (ingredients: Chaihu 20 g, huangqin 10 g, banxia 15 g, gancao 10 g) and group Ⅱ (ingredients: Chuanqiong 20 g, tianma 15 g, xixin 5 g, quanxie 5 g, wugong 5 g): Gastric perfusion of 0.75 g/mL agents of treatment with DS and DW wind antidyne was given to rats as 8.70 g/kg; Normal control group: Gastric perfusion of normal saline of the same volume wasgiven to rats.Intervention on rats of all groups lasted 12 days.② Experiment of thrombogenesis in vitro: The cephalic artery of one side was seperated 2 hours after the last time of administration. The distal part was deligated, the blood current in proximal part was blocked with bulldog clamp and the arterial cannula was inserted into the cephalic artery. Loosened the bulldog clamp, blood of 1.8 mL was collected and put into the rotary ring of thrombogensis meter to rotate for 15 minutes at 17 r/min,and then poured the thrombus, measured the length and humid weight of thromb. After that,dried the humid thromb and measured the dry weight.③ Determination of PA:The packing fraction (PF) in the 1st and 5th minutes as well as the maximal packing fraction (MPF) and assembling inhibition ratio of platelet-rich plasma within 5 minutes were recorded with test.MAIN OUTCOME MEASURES: Effect of treating Shaoyang disease nephelometry.④ Differences of measurement data were compared with tand methods of DW as well as RP on thrombogenesis in vitro and the function of PA.RESULTS: A total of 60 rats were involved in the analysis of results. ①The length, humid weight and dry weight of thrombogenesis in vitro in group Ⅲ and Ⅳ, positive control group and group Ⅱ were obviously lower than those in group Ⅰ (P ＜ 0.05). ② The PF in 1st, 5th minute and the maximal aggregathe normal control group (P ＜ 0.05-0.01); The humid weight and dry weight of thromb in group Ⅳ were significantly lower than those in tion (MA) of blood platelet in rats of each group were significantly lower than those in the normal control group (P ＜ 0.05-0.01), and the curative effect on rats in the positive control group, group Ⅲ and Ⅳ as well as group Ⅱ were better than that in group Ⅰ.CONCLUSION: There are no significant effects of dispersing thrombogenesis in vitro and inhibiting PA by only treating Shaoyang disease. The effect of combined prescription of DW and RP as well as treatment of Shaoyang disease are obviously enhanced.

Objective To investigate the effect of using prophylactic antidepressant treatment with citalopram on motor function recovery in patients with acute cortical infarction. Methods The patients with first-ever acute middle cerebral artery territory cortical infarction and with upper limb motor deficits within 24 h after onset were enrol ed prospectively. They were randomly divided into either a citalopram group or a control group. On the basis of conventional treatment, the citalopram group was given citalopram hydrobromide tablets oral y (20 mg/d) within 48 h after onset of stroke for 30 d. The 17-item Hamilton Depressive Rating Scale (17-item HDRS) was used to conduct the assessment of depressive symptoms before and after treatment. The National Institutes of Health Stroke Scale ( NIHSS ) and Fugl-Meyer Assessment (FMA) hand motor function score were used to conduct function assessment before and after treatment. Results A total of 66 patients were enrol ed in the analysis, including 32 patients in the citalopram group and 34 in the control group. There was no significant difference in baseline clinical data between the 2 groups. The baseline stroke severity of al patients was moderate (NIHSS score 5-15). Compared with baseline level at day 30 after the treatment, the HDRS score of the citalopram group was improved significantly (6. 70 ± 3. 58vs. 9. 86 ± 3. 48; t= 3. 380, P= 0. 001 ). However, there was no significant difference with the control group after treatment (6. 70 ± 3. 58 vs. 8. 12 ± 2. 96;t=1. 745, P=0. 086). The NIHSS scores in the citalopram group and the control group were decreased respectively (4. 30 ± 1. 88 vs. 2. 00 ± 1. 24;t=5. 900, P<0. 001). The hand FMA scores were increased significantly (4. 00 ± 0. 70 vs. 1. 42 ± 1. 91;t=7. 197, P<0. 001). There was significant difference between the 2 groups. In addition, the hand motor function score at day 30 after treatment was more than 25% of the baseline recovery in 14 patients of the citalopram group (43. 8%) and in 5 of the control group (14. 7%). The citalopram group was significantly higher than the control group (χ2 =6. 783, P=0. 009 ). Conclusions Early prescription of citalopram can significantly promote the motor function recovery in acute cortical infarction patients with moderate motor deficit and its mechanism may not be associated the antidepressant effect.

BACKGROUND:Studies have shown that apoptosis is closely related to the pathogenesis of steroid-induced femoral head necrosis. The release of cytochrome c plays a very important role in the process of apoptosis.
OBJECTIVE:To observe the effects of cytochrome c on early steroid-induced femoral head necrosis.
METHODS:Twenty-four healthy adult male New Zealand rabbits at 5 months old were randomly divided into model group and control group (n=12 per group). Models of early steroid-induced femoral head necrosis were established by intragluteal injection of hormone combined with ear vein injection of horse serum. In the control group, rabbits were given ear vein injection of the same amount of physiological saline. At 2, 4 and 6 weeks after model establishment, histopathological changes of bilateral femoral head were observed by optical microscope, and the ratio of empty lacuna was calculated. Apoptosis of osteocytes was determined by TUNEL assay, and apoptotic index was calculated. Immunohistochemical method was used to determine cytochrome c and to calculate cytochrome c-positive expression rate.
RESULTS AND CONCLUSION:(1) The ratio of empty lacuna and apoptotic index:The model of early steroid-induced femoral head necrosis was successful y established in the experiment. Compared with the control group, ratio of empty lacuna, apoptotic index and expression rate of cytochrome c in osteocytes were significantly higher in the model group (P<0.05 or P<0.01). (2) Correlation analysis:Ratio of empty lacuna was significantly positively associated with apoptotic index at various time points in the model group (r=0.856, P<0.01). Expression rate of cytochrome c was significantly positively associated with apoptotic index at various time points in the model group (r=0.824, P<0.01). (3) These findings confirm that cytochrome c-involved apoptosis of osteocytes may play an important role in steroid-induced femoral head necrosis. Expression rate of cytochrome c in osteocytes is remarkably positively associated with the occurrence of steroid-induced femoral head necrosis in rabbits.

BACKGROUND:Increasing evidence has demonstrated that bone marrow mesenchymal stem cel (BMSC) transplantation can promote skin, liver and lung repair in animal models; andLeonurus sibiricus L. has the ability of promoting blood circulation and regulating menstruation.
OBJECTIVE: To investigate the therapeutic effect of BMSC transplantation with leonurine on intrauterine adhesions (IUA) in rabbits and the relevant mechanism of action.
METHODS: After modeled using dual injury method, rabbit models of IUA were randomly divided into five groups: sham-operated group (sham), model group (IUA), BMSC transplantation group, leonurine treatment group and combined treatment group (BMSCs+leonurine). Rabbits in the sham group were only given normal saline rinsing after hysterotomy, while those in the latter three groups were correspondingly given intrauterine BMSC transplantation or/and intragastric administration of 4 mg/kg leonurine for 14 days. Morphological changes of the endometrium were observed using hematoxylin-eosin staining, and expression levels of transforming growth factor β protein, Smad3 protein and interferon-γ mRNA were detected using immunohistochemical staining and real-time fluorescence quantitative PCR, respectively.
RESULTS AND CONCLUSION:Compared with the model group, the degree of IUA was al significantly improved in the other groups, especialy in the combined treatment group. Moreover, BMSC transplantation, leonurine treatment and their combined use al could inhibit IUA-induced increase of transforming growth factorβ and Smad3 protein expression and IUA-induced decrease of interferon-γ mRNA level. Importantly, al these alternations were much more pronounced in the combined treatment group. Our results show that the combined use of BMSC transplantation and leonurine treatment can exert a synergistic effect in the improvement of IUA through the transforming growth factor β/Smad3 pathway.

The determination of fraxetin A and fraxetin B in Cortex Fraxini have been carried out by HPLC on the stationary phase:octodecyl-silicohydride bonded silica gel column. The mobile phase was methanol-water(24:76). The detection wavelength was at 348nm. There was a good linear relationship in a concentration range of 16?g/ml~176?g/ml. The correlation coefficients were 0. 9995 and 0. 9994,respectively. The recovery of the added sample were 99. 1 ?1. 5% (n=5) for fraxetin A and 99.8?2. 5%(n=5) for fraxetin B. This method showed 0. 8% of within-day accuracy and 6. 2% of between-day accuracy,involving the advantages of simplicity,quickness and accuracy.

Objective: To establish the identification of protein compositions in Rhizoma Arisaematis through researches of its taxonomy, pharmacognostic anatomy and chemical compositions.Methods: The protein compositions in Rhizoma Arisaematis were identified by HPCE taking potassium phosphate boric acid buffer solution (pH=9.0) as an electrophoretic buffer. The electric sampling time was 10s, 10kV; Separating pressure 25kV; column temperature=28℃;?=205nm; integral determination sensitivety value=0.05; sensitivity=0.1 AUFS. Also, the raw plant identification, medical material characteristics and microscope identification were performed. Results: Combined with pharmacognostic and microscopic identification, experiments indicated that herbs of Arisaematis genus have similar HPCE chromatographic profile but different finger peaks. Condlusion: This study provides a better basis for the identification of Rhizoma Arisaematis.

At present, the roles of neural plasticity and motor functional recovery in stroke have increasingly been paid attention to. The remodeling of cerebral cortex after stroke can complement the function of injured tissues, which has been confirmed by animal experiments. Functional neuroimaging technique has been gradually used in the evaluation of neural plasticity.

0.05),but existed between grades Ⅰ-Ⅲ and grade Ⅳ(P0.05).Conclusion PIAS3 is expressed in human gliomas,closely correlated with the pathological grade and malignant intensity.PIAS3 may be involved in the development and progression of gliomas.