An HPLC method for the quantitative determination of icariin and an UV method for the determination of flavonoids in Co-Wuzi granule Preparation were developed. Results showed that the amount of icariin was 0. 050%, RSD=0. 72%, and the amount of flavonoids was 0. 120%, RSD=3. 8%. The HPLC method was very effective in the separation of active principles,suitable for the quantitative determination of single component in compounded TCM preparations,while the UV method may be used for the identification of active parts of TCM preparations.

Objective To explore the adeB gene expression level impact of tigecycline for multidrug resistance Acinetobacter baumannii.Methods 57 strains multidrug resistance Acinetobacter baumannii were separated from September.2012 to September.2014.Epsilome test method was used to detect minimum inhibitory concentration of tigecycline against multi drug resistant Acinetobacter baumannii.PCR amplification detected adeB, adeR, adeS efflux pump gene distribution.Real-time fluorescent RT-PCR detection technology were used to detect relative expression level of each gene. Results 57 strains of multidrug-resistant Acinetobacter Bauman MIC value were 0.1-8μg/mL.The calculation of MIC50 =2μg/mL, MIC90 =4μg/mL. 49 strains sensitivity accounted for 86.0%, 5 intermediate strains accounted for 8.8%, 3 resistant strains accounted for 5.2%.57 strains of bacteria were divided into 38 strains lower sensitivity group and 19 strains sensitive group, with reduced susceptibility in lower sensitivity group 30 strains, accounting for 78.9%, sensitive group were all sensitive to tigecycline, accounting for 100%, for tigecycline susceptibility sensitive group was significantly higher than that of lower sensitivity group ( P <0.65 ) .41 strains were detected adeB, adeR, adeS gene in all strains, accounted for 86.0%, 29 strains were detected adeB, adeR, adeS genes in lower sensitivity group, accounted for 70.7%,12 strains were detected in sensitive group accounted for 63.2%, the difference between the two groups had no statistical significance.Fluorescent RT-PCR detection results showed that the relative expression of adeB gene with lower susceptibility group was significantly lower than that of sensitive group (P<0.05), relative expression levels of adeR and adeS gene in two groups had no significant difference.Conclusion AdeB gene plays an important role in tigecycline for multidrug resistance Acinetobacter baumannii, but may also exist outside of the adeB gene is involved in the regulation of resistance to tigecycline.

Objective:To observe the clinical efficacy of linear scalp-acupuncture combined with herbs in the treatment of Parkinson's disease(PD).methods:One hundred and fifty PD cases were randomized into four groups in the sequence ofvisit.Eighty-eight cases in group A,were treated by linear scalp.acupuncture and herbs effective to nourish the liver and kidneys,activate blood and remove wind,and acupuncture was performed every other day and 10 treatments made uP one course;20 cases in group B,were treated only by linear scalp-acupuncture;20 cases in group C,were treated by herbs effective to nourish the liver and kidneys,activate blood and remove wind;22 cases in group D,were treated by Dopamine.All groups were treated for three months.Resuits:The total response rate was 89.8%in group A,similarto 86.4%in group D(P＞0.05),and higherthan 65.0%in group C(P＜0.01).Conclusion:Linear scalp-acupuncture combined with herbs is quite effective in the treatment of PD.

Objective To evaluate the effect of intrathecal methotrexate on activation of spinal astrocytes in a rat model of bone cancer pain (BCP).Methods Sixty female unmated Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),BCP group (group P),and BCP + methotrexate group (group PM).BCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.On day 7 after BCP,methotrexate 100 μg (diluted to 15 μl in artificial cerebrospinal fluid) was injected intrathecally over 10 s in group PM,and artificial ccrebrospinal fluid 15 μ1 was injected intrathecally over 10 s in S and P groups.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before BCP and 3,7,14 and 21 days after BCP.Five rats were sacrificed after measurement of the pain threshold at 7,14 and 21 days after BCP,and the lumbar enlargement segments of the spinal cord were harvested for detection of the expression of glial fibrillary acidic protein (GFAP) by immuno-histochemistry.Five rats were sacrificed after measurement of the pain threshold at 14 days after BCP,and the expression of GFAP in the spinal cord was detected by Western blot.Results Compared with group S,the MWT was significantly decreased at 3,7,14 and 21 days after BCP in C and CM groups,the expression of GFAP was significantly up-regulated at each time point after BCP in group C,and the expression of GFAP was significantly up-regulated at 7 and 14 days after BCP in group CM (P＜0.05).Compared with group C,the MWT was significantly increased,and the expression of GFAP was significantly down-regulated at 14 and 21 days after BCP in group CM (P＜0.05).Conclusion The mechanism by which intrathecal methotrexate reduces BCP may be related to inhibition of spinal astrocyte activation in the rats.

[Objective] To establish an effective way for the detection of Huanglongbing pathogen in Citrus medica L. var. sarcodactylis (Noot. ) Swingle by polymerase chain reaction (PCR) analysis, and to provide evidence for the early diagnosis of the culture of pathogen-free plantlets and for the prevention and control of diseases. [ Methods ] The incidence of Huanglongbing disease was investigated in the main producing area of Citrus medica L. var. sarcodactylis (Noot.) Swingle. By using the special primers designed by the DNA sequence of Citrus Huanglongbing pathogen, PCR amplification of DNA from the samples was conducted. [Results] Symptoms similar to the Citrus Huanglongbing disease occurred in Citrus medica L. var. sarcodactylis (Noot.) Swingle growing in Zhaoqing, Guangdong. The results of PCR analysis showed that a special DNA fragment of 400bp had been detected in the affected plants of Citrus medica L. var. sarcodactylis (Noot. ) Swingle, but it was not found in the healthy plants, indicating that the symptoms were caused by the Huanglongbing pathogen. [ Conclusion ] PCR analysis is an effective way for the detection of Huanglongbing pathogen in Citrus medica L. var. sarcodactylis ( Noot. ) Swingle.

Objective To study the state of doctors' psychological capital in public hospitals,and to analyze the characteristics of such capital,so as to better leverage the capital as a hospital management tool.Methods A quantitative and qualitative research was made on the state of doctors＇ psychological capital.Results The doctors' task-oriented psychological capital scored 4.25,and guanxi-oriented psychological capital 4.41; “modesty and prudence”scored the highest in all dimensions of the doctors' psychological capital,“optimism and hope”lowest; the doctors with different genders,ages,seniority,and monthly average income have a difference in psychological capital.However,no significant difference was found with varying titles,hospital levels,and employment models.Conclusion The doctors were found with a strong psychological capital to cope with stress.Better psychological capital will raise quality of care in practice.

Objective To investigate the association between the apolipoprotein E (apoE) gene polymorphism and the dose for warfarin individual maintenance. Methods The genotypes of 249 patients with warfarin treatment in maintenance doses were determined by PCR/DHPLC assay. The doses for warfarin maintenance were compared among patients with different genotypes. Results In the total of 249 patients, the frequencies of 2/ε2, ε2/ε3, ε2/ε4, ε3/ε3, ε3/ε4, ε4/ε4 genotype were 1.20%, 15.66%, 1.80%, 72.29%, 9.24%, 0.80%, respectively; the allele frequencies of ε2, ε3, ε4 were 9.44%, 84.74%, 5.82%, respectively. The warfarin dose of group ε2 (ε2/ε2, ε2/ε3) was (3.24±1.36) mg/d, slightly higher than that of group ε3 (ε3/ε3, 2.91±1.14 mg/d) or group ε4 [ε4/ε4, ε3/ε4, (2.98±1.05) mg/d], but the difference of the warfarin doses among the 3 groups did not reach statistical significance (F=1.848,P＞0.05). Conclusion ApoE polymorphism may be not a major genetic factor that influences the individual dose for warfarin maintenance.

BACKGROUND:Extraction methods of human amniotic mesenchymal stem cells are inconsistent in the number of cells.
OBJECTIVE:To explore the optimal method to in vitro isolate and culture human amniotic mesenchymal stem cells.
METHODS:Under sterile conditions, ful-term cesarean fetal amniotic membrane was cut into pieces, then to isolate human amniotic mesenchymal stem cells by seven methods in four experiments. In experiment 1, human amniotic mesenchymal stem cells were isolated by the fol owing three methods:(1) 0.05 g/L trypsin digestion for 10 minutes fol owed by 0.75 g/L col agenase digestion for 60 minutes;(2) 0.75 g/L col agenase I for 120 minutes;(3) co-digestion with 0.05 g/L trypsin and 0.75 g/L col agenase for 60 minutes. In experiment 2, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes fol owed by 0.75 g/L col agenase digestion for 30 minutes. In experiment 3, the samples were digested by two methods:(1) 0.05 g/L trypsin digestion for 30 minutes×2, fol owed by 0.75 g/L col agenase digestion for 60 minutes;(2) 0.05 g/L trypsin digestion for 40 minutes×2, fol owed by 0.75 g/L col agenase digestion for 60 minutes. In experiment 4, the samples were digested with 0.05 g/L trypsin digestion for 30 minutes×2, fol owed by 1 g/L col agenase digestion for 60 minutes. Fol owing morphology observation under a microscope, we studied the most suitable method for isolating human amniotic mesenchymal stem cells.
RESULTS AND CONCLUSION:Digestion with 0.05 g/L trypsin for 30 minutes twice fol owed by 1 g/L of col agenase digestion of 60 minutes was the most suitable isolation and culture condition in vitro. cells became elongated fusiform or star-shaped with rich cytoplasm, and nuclei were round with 1-3 nuts. We can harvest the most number of human amniotic mesenchymal stem cells using the method described in experiment 4.

ObjectiveToinvestigatetheeffectandmechanismofaChinesetraditionalprescription,YuebiBanxia decoction, on allergic asthma in mice by observing the changes of serum immunoglobulin E ( IgE ) and lung tissue superoxide dismutase ( SOD) activity in the mouse model of allergic asthma .Methods Forty healthy Kunming mice were randomly divided into normal control group , asthma model group , dexamethasone intervention group and Chinese medicine preparation group .The mouse model of asthma was generated by peritoneal injection of ovalbumin ( OVA) and suspension of aluminium hydroxide .The mice of traditional Chinese medicine group received Yuebi Banxia decoction in a dose of 1 mL/100 g, daily for 30 days.The mice of dexamethasone group were given intraperitoneal injection of dexamethasone 1 mg/kg.The normal control group and asthma model group were given gastric gavage of physiological saline.After 30 days administration, the serum levels of IgE and SOD activity of lung tissue were determined , and the pathological changes of lung tissue were observed using HE staining .Results The mice of traditional Chinese medicine preparation group showed significantly decreased serum IgE level , and significantly increased SOD activity than those of the asthma model group mice (P<0.05 for both).Conclusions Reduced serum IgE and increased SOD activity in the lung tissue of mice with allergic asthma , improving the antioxidant ability of lung tissue , may be involved in the anti-allergic asthma mechanism of the Chinese traditional prescription Yuebi Banxia decoction .

OBJECTIVE:To establish a method for the simultaneous determination of 5 unsaturated fatty acids in Perilla oil cap-sule. METHODS:With the reference material of α-linolenic acid methyl ester,GC was used to determine and calculate the relative correction factors of α-linolenic acid methyl ester with methyl palmitate,methyl stearate,methyl oleate and linoleic acid methyl es-ter,and the correction factors were used to calculate the contents of 5 unsaturated fatty acids;the column was Agilent Innowax cap-illary column,the detector was FID,the inlet temperature was 230 ℃,the detector temperature was 250 ℃,the gas flow rate was 20 ml/min(nitrogen),40 ml/min(hydrogen)and 350 ml/min(air),split ratio was 30 to 1,the column temperature was 190 ℃, and injection volume was 1 μl. RESULTS:The linear range was 0.018-0.792 μg(r=0.9994)for methyl palmitate,0.0016-0.0176μg(r=0.9993)for methyl stearate,0.0056-0.2464 μg(r=0.9999)for methyl oleate,0.003-0.132 μg(r=0.9990)for linoleic acid methyl ester and 0.018-0.792 μg(r=0.9998) for α-linolenic acid methyl ester;RSDs of precision,stability and reproducibility tests were lower than 5%;recoveries were 98.990%-101.70%(RSD=0.720%,n=6) for methyl palmitate,99.599%-100.699%(RSD=0.368%,n=6) for methyl stearate,98.996%-101.680%(RSD=1.240%,n=6) for methyl oleate,99.813%-100.963%(RSD=0.434%,n=6)for linoleic acid methyl ester and 97.185%-99.602%(RSD=0.874%,n=6)for α-linolenic acid methyl es-ter. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determina-tion of methyl palmitate,methyl stearate,methyl oleate,linoleic acid methyl ester,α-linolenic acid methyl ester in Perilla oil cap-sule.