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 explore the best humidifying methods of artificial airway by comparison of different humidifying approaches.Methods 124 patients were divided into the control group (60 cases) and the treatment group (64 cases).Two methods,continuous airway humidifying by the trace injection pump and continuous airway humidifying by oxygen atomization were adopted.The humidifying effect,tolerance rate of patients,the airway complications and nursing cost were compared between the two groups.Results The method of continuous airway humidifying by oxygen atomization had better effect,was easy to be tolerated,less complications and less cost.Conclusions Continuous airway humidifying by oxygen atomization is a safe,effective,low-cost method,it is easy to be tolerated by patients and is worthy of being popularized.

Objective The effects of nemo-like kinase(NLK)in inflammation of central nervous system were explored. Methods An animal model with central nervous system inflammation disease induced by intracerebroventricular infusion of lipopolysaccharide was constructed. A model of neuronal apoptosis induced by glutamate in PC12 cells was constructed. A pcdna 3.1-NLK over-expression plasmid and a pSilencer 4. 1-CMV-NLK small interfering plasmid were also constructed. The expression and location of NLK were detected using western blot analysis, double immunofluorescent staining, and cell counting kit-8 assay. Results The protein level of NLK was reduced after induction of inflammation in the brain(t =2. 718-3. 106, all P ＜0. 01), and NLK was only expressed in both cortical and hippocampal neurons. At the cellular level, NLK expression was gradually reduced along with neuronal apoptosis induced by glutamate in PC12(t =4. 032, P ＜0. 01). Over-expression of NLK would reduce the apoptosis of neurons induced by glutamate(t =3. 930, P ＜ 0. 01). Conversely, interfering the expression of NLK would enhance neuronal apoptosis(t = 2. 845, P ＜ 0. 01). Conclusion NLK can protect neurons by inhibiting apoptosis in the process of central nervous system inflammation.

5-fluorouracil (5-Fu) as an antimetabolite drug,has been widely used in the treatment of various solid tumors.However, due to the heterogeneity of tumor and the impact of the individual, some patients still have drug resistance or serious lethal toxicity reaction.The text focuses on the research progress of pharmacogenomics and pharmacogenetics of some forecast elements which affect the efficacy and toxicity of 5-Fu,and discussed their guiding role on the choice of chemotherapy drugs.

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.

Activation of HGF/Met has been complicated in tumorigenesis, invasion and metastasis of various tumors. We summarize current knowledge on their association with invasion of thyroid carcinoma. By discussing the expression of Met and its possible mechanism and analyzing the activation of Met with tumor motility, angiogenesis and inflammatory reaction, we conclude that the research of the possible role of HGF/Met in thyroid carcinoma will advance knowledge on the diagnosis and management of this disease,as well as tumor invasion.

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:The etiological factor of pediatric avascular necrosis of the femoral head(Legg-Calv?-Perthe disease) is unclear right now,and the disease typing,treatment and prognosis outcome are also different.Thus a simple,practical and reliable clinical classification is necessary to guide treatment and estimate result.OBJECTIVE:To summarize and analyze the criteria commonly used for Legg-Calv?-Perthe disease in clinical typing as well as their advantage and shortcoming,which provides evidences for the treatment and instruction.RETRIEVAL STRATEGY:A computer-based search of Medline database was undertaken to identify the articles regarding the classification criteria for Perthes disease,published between January 1970 and September 2007,with the key items of "Perthes,classification" in English.After the primary check,the full-texts of the remained articles were looked for.Inclusion criteria:①X-ray,bone scanning and MRI classification criteria for Perthes disease.②the study of the correlation between the classification reliability and the prosthesis.Exclusion criteria:repeated researches.Totally 76 articles about the classification criteria for Perthes disease were retrieved,and 32 of them were in accordance with the inclusion criteria.LITERATURE EVALUATION:Among 32 included articles,24 ones concerned with X-ray examination,4 with bone scanning image,and 4 with MRI image.DATA SYNTHESIS:There were many related researches to the clinical classification of Legg-Calv?-Perthe disease,but uniform conclusion is absent.A reliable and unified clinical classification is beneficial for the orthopaedic physicians to find out the risk factors at early stage,and take therapeutic measures as soon as possible,so as to prevent the malformation of the femoral head.Ideal classification criteria with good reliability and consistency,are closely related to the disease prosthesis,and should be used at early stage.CONCLUSION:X-ray typing is commonly used in the clinical classification for Legg-Calv?-Perthe disease,but it is difficult to apply at early stage.Bone scanning can forecast results,and also induce irradiation.Due to good sensitivity and high resolution,MRI has an earlier effect on detecting necrosis and collapse,and is potential for further development and application,however,it also calls on a long-term clinical follow-ups.