Objective:To develop a new preparation process of PLGA microparticles for protein drugs by SPG membrane emulsifi-cation combined with W/O/W double emulsion-solvent technique. Methods:Lysozyme was used as the model drug to prepare the mi-croparticles. The influence of formula factors on the properties of the microparticles was studied, and the physicochemical properties, in vivo compatibility and degradation of the microparticles were investigated as well. Results:The drug loading of lysozyme-loaded mi-croparticles was 35%, the entrapment efficiency was 72. 43% and the average size was 63. 89 μm with PDI of 0. 675. DSC and FTIR showed that lysozyme was entrapped in the microparticles. The microspheres had promising biocompatibility and sustained degradation in vivo. Conclusion:The paper describes a new satisfactory preparation process of PLGA microparticles for protein drugs with good in vitro and in vivo properties.

AIM:To investigate the effect of Ginkgo biloba extract(EGb761) on cell proliferation and expression of transforming growth factor ?1(TGF-?1) and connective tissue growth factor(CTGF) in rat hepatic stellate cells(HSC-T6).METHODS:HSC-T6 was cultured with EGb761 at concentration of 1,10,100 and 500 mg/L,and without EGb761 as blank control.MTT colorimetric assay was used for detecting the proliferation of HSCs and flow cytometry was used for observing the cell cycles of HSCs under different concentrations of EGb761.The expression of TGF-?1/CTGF mRNA and protein levels were determined by reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting after 24 hours and 48 hours.RESULTS:EGb761 inhibited the proliferation of HSCs at the concentration of 10 mg/L,100 mg/L and 500 mg/L.G0/G1 stage was increased(P

Yishen Juanbi Method for Treating Rheumatism began from Mr. ZHANG Ci-gong to the formation and development of Mr. ZHU Liang-chun, and by the evidence-based source, innovation and perfection of the third and fourth generations of successors, has formed the unity of the principle, rule, method, medicine, and the unique diagnosis and treatment of rheumatism of integration of the internal treatment and external treatment. All of these provide original insights for rheumatology of TCM theory and the application of insect medicine, and provide valuable treatment technology for China's young rheumatology disciplines.

Objective To develop a method for rapid detection of Mycoplasma pneumoniae and its macrolide resistance mutation. Methods The primers and cycling probe sets were designed to detect two single nucleotide mutation, A2063G and A2064G, in the 23s rRNA gene of Mycoplasma pneumoniae. By using recombinant plasmids containing 23s rRNA gene fragments, 102 Mycoplasma pneumoniae clinical isolates from 2005 to 2008, and 136 nasopharyngeal suction specimens from pediatric patients with low respiratory tract infections in Shanghai Children's Hospital from November to December in 2009 were investigated to determine the specificity and the sensitivity of the CycleavePCR method. PCR amplification and sequence analysis of 23S rRNA genes were performed for all Mycoplasma pneumoniae strains and Mycoplasma pneumoniae positive specimens to confirm the results of the CycleavePCR method. Results Of 102 clinical isolates, 83 was resistant to erythromycin and sequence results show that all macrolide-resistant Mycoplasma pneumoniae strains harbored an A2063G ( 82/83 ) or A2064G ( 1/83 ) transition mutation in 23S rRNA genes. Twelve was Mycoplasma pneumoniae detected positive by CycleavePCR in 136nasopharyngeal suction specimens. The CycleavePCR results were consistent with those of routine PCR and sequencing. There was no signal production from other bacterial species. Sensitivity and specificity were 100%. The detection limit of the CycleavePCR was 10 plasmid copies per reaction. Experiment can be done within 1.5 h. Conclusion A novel method is developed to detect erythromycin-resistant strains harboring A2063G and A2064G transition mutation in the 23s rRNA gene using CycleavePCR.

Objective To learn the current in vitro antimicrobial susceptibility of Mycoplasma pneu-moniae in Shanghai and to understand the mechanisms of resistance to macrolides. Methods M. pneumoniae was isolated from pediatric patients with low respiratory tract infections(RTI) using broth and PPLO agar medi-um. PCR amplification and sequence analysis of P1 adhesion gene were performed to identify all M. pneumoniae strains. Susceptibility testing was carried out for macrolides, tetracyclines and fluoroquinolones using broth mi-crodilution method with SP4 broth. PCR amplification and sequence analysis of 23S rRNA genes were performed for all M. pneumoniae strains. P1 gene PCR-RFLP typing was performed to subtype the M. pneumoniae strains. Results One hundred and two M. pneumoniae strains were isolated in Shanghai from Oct 2005 to Dec 2008. All M. pneumoniae isolates were susceptible to the tetracyclines and fluoroquinolones tested. Of 102 clinical isolates, 83(81.4%) was resistant to erytbromycin and all 83 erythromycin-resistant strains had MIC>128 mg/L. An increasing trend of resistance rates were showed: 16.7% (1/6) in 2005, 76.5% (13/17) in 2006, 100.0% (24/24) in 2007 and 81.8% (45/55) in 2008. All macrolide-resistant M. pneumoniae strains harbored an A2063G transition mutation in domain V of 23S rRNA genes. The P1 gene RFLP type 1 is predominant (85.3%, 87/102) in M. pneumoniae clinical isolates. Conclusion The macrolide resistance rate of M. pneu-moniae is very high in Shanghai. The mechanism of macrolide resistance is associated with transition mutation on the 23S rRNA gene.

Objective To explore the clinical head nurses'effective competence through interviewing nursing supervisors by in depth interview method and to provide reference for constructing training content for clinical head nurses in Macau and Guangzhou.Methods In depth interview was carried out among 8 nursing supervisors from 2 hospitals in Macau and 9 nursing supervisors from 2 tertiary hospitals in Guangzhou.Content analysis was utilized through organizing records and then forming written documentations.Results The themes were drawn after the interview,which were good moral character accomplishment,good communication skill,prospective and creative thinking,favorable executive ability.Forwardlooking and innovative were significantly different in Macau and Guangzhou.Conclusions The effective clinical head nurses' competence is multi-faceted,the results can provide reference for selection,evaluation and training for head nurses.

OBJECTIVETo document morphological changes in hepatic microcirculation in liver tissue with hepatitis B and the pathogenesis of hepatic microcirculatory disturbances.

METHODSLiver tissue samples were obtained from patients with hepatitis B by liver biopsy. These samples were examined with a light microscope and transmission electron microscope.

RESULTSHepatic microcirculatory disturbances existed in patients with hepatitis B, including those with normal liver function, manifested by red blood cell aggregation in sinusoids seen under light microscope and sinusoidal capillarization seen under electron microscope. Weibel-Palade bodies in sinusoidal endothelial cells were seen in 26 out of 53 cases. Intimate contacts were found between lymphocyte/Kupffer cells and sinusoidal endothelial cells.

CONCLUSIONSHepatic microcirculatory disturbances exist in patients with hepatitis B. The appearance of Weibel-Palade bodies in sinusoidal endothelial cells may be a key step in the development of hepatic microcirculatory disturbances.

Adolescent ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; pathology ; physiopathology ; Humans ; Liver ; blood supply ; pathology ; ultrastructure ; Liver Circulation ; Male ; Microcirculation ; pathology ; ultrastructure ; Microscopy, Electron ; Middle Aged

OBJECTIVE: To analyze the indications of the therapies for rheumatoid arthritis (RA) with neural network model analysis. METHODS: Three hundred and ninety-seven patients were included in the clinical trial from 9 clinical centers. They were randomly divided into Western medicine (WM) treated group, 194 cases; and traditional Chinese herbal medicine (CM) treated group, 203 cases. A complete physical examination and 18 common clinical manifestations were prepared before the randomization and after the treatment. The WM therapy included voltaren extended action tablet, methotrexate and sulfasalazine. The CM therapy included Glucosidorum Tripterygii Totorum Tablet and syndrome differentiation treatment. The American College of Rheumatology 20 (ACR20) was taken as efficacy evaluation. All data were analyzed on SAS 8.2 statistical package. The relationships between each variable and efficacy were analyzed, and the variables with P<0.2 were included for the data mining analysis with neural network model. All data were classified into training set (75%) and verification set (25%) for further verification on the data-mining model. RESULTS: Eighteen variables in CM and 24 variables in WM were included in the data-mining model. In CM, morning stiffness, swollen joint number, peripheral immunoglobulin M (IgM) level, tenderness joint number, tenderness, rheumatoid factor (RF), C-reactive protein (CRP) and joint pain were positively related to the efficacy, and disease duration and more urination at night negatively related to the efficacy. In WM, erythrocyte sedimentation rate (ESR), weak waist, white fur in tongue, joint pain, joint stiffness and swollen joint were positively related to the efficacy, and yellow fur in tongue, red tongue, white blood negatively related to the efficacy. In the analysis with the neural network model in the patients of verification set, the predictive response rates of 20% patients would be 100% and 90% in the treatment with CM and WM, respectively. CONCLUSION: Neural network model analysis, based on the full clinical trial data with collection of both traditional Chinese medicine and modern medicine diagnostic information, shows a good predictive role for the information in the efficacy in rheumatoid arthritis.

OBJECTIVE: To evaluate the correlations between clinical symptoms and treatment efficacy in patients with rheumatoid arthritis (RA). METHODS: Four hundred and thirteen patients were included in the clinical trial from 9 clinical centers. They were randomly divided into Western medicine-treated group with 204 cases and Chinese herbal drug-treated group with 209 cases. Eighteen clinical symptoms were evaluated before and after treatment. The Western medicine therapy included voltaren extended release tablets, methotrexate and sulfasalazine. The Chinese herbal drug therapy included glucosidorum Tripterygii totorum tablets and Yishen Juanbi Tablets combined with treatment based on syndrome differentiation. The American College of Rheumatology 20 (ACR20) was used as efficacy evaluation criteria. RESULTS: In the Chinese herbal drug-treated group, clinical symptoms such as arthralgia and tenderness of joints were positively correlated with the efficacy after 12-week treatment, while frequent urination at night was negatively correlated. In the same group, tenderness of joints and fever were positively correlated with the efficacy after 24-week treatment, while deep-colored and turbid urine was negatively correlated. In the Western medicine-treated group, tenderness of joints and thirst were positively correlated with the efficacy after 12-week treatment, while vertigo was negatively correlated. And in the same group, tenderness of joints was positively correlated with the efficacy after 24-week treatment, while heaviness of limbs was negatively correlated to the efficacy. The statistical results showed that the treatment efficacy was improved when the correlated symptoms were included in the indications. CONCLUSION: The treatment efficacy of RA is correlated with some symptoms, so further studies should proceed on these correlations in order to achieve better treatment outcome.

Objective To assess the cerebrovascular reserve (CVR)and the relationship of CVR with the short-term prognosis in patients with acute atherosclerotic cerebral infarction.Methods A total of 106 patients with unilateral acute (within 72 h) atherosclerotic cerebral infarction (trial group) were selected from December 2016 to December 2017 in the Department of Neurology of the First People's Hospital of Hefei,which were divided into two groups including the lesion group (106 cases) and the nonlesion group (106 cases).The median score of NIHSS in patients was 4(2,5).The control group included 40 healthy controls.The cerebral blood flow reserve and pulsatility index were measured by transcranial Doppler combined with CO2 inhalation test in both the trial group and the control group.According to the rate of change of cerebral blood flow velocity (CBFV),all subjects were divided into two groups including the normal group and the impaired cerebral blood flow reserve group.The changes of CBFV were compared in the control group and the trial group,which was divided into two groups including the group with lesion side and the group with non-lesion side.To evaluate the brain structure reserve the circle of Willis in the trial group was assessed by MRA.According to the integrity of the circle of Willis anterior and posterior circulation all subjects were divided into four groups (type Ⅰ,type Ⅱ,type Ⅲ and type Ⅳ).The effect of the factors,such as diabetes,hypertension,low density lipoprotein (LDL),high density lipoprotein (HDL),smoking,and drinking history,on cerebral blood flow reserve was measured by single-factor analysis.The correlation of NIHSS scores,infarct size and volume with CVR was also measured.All patients in the trial group were treated with drugs and were followed-up for three months.The modified Rankin Scale (Mrs) was used to evaluate the prognosis of the patients.It means poor prognosis if the value of Mrs was more than three.The effects of factors,such as sex,HDL,LDL,diabetes,hypertension,smoking history,drinking history,cerebral blood flow reserve,NIHSS scores,brain structure reserve,infarct location,age,on the prognosis were measured by multivariate Logistic regression.Results The increase rate of CBFV in the lesion-side of patients with atherosclerotic cerebral infarction was 5.94％ (2.18％,10.49％),and the increase rate of pulsatility index was 10.77％ (2.21％,22.62％),which were both lower than the control group (CBFV:11.54％ (5.01％,17.96％),Z =2.547,P＜0.05);pulsatility index:48.36％ (33.93％,64.51％),Z =6.604,P ＜ 0.01).There was significant difference (x2 =4.328,P ＜ 0.05) in the distribution of diabetes,which was 2/14 in the normal group and 43.48％ (40/92) in the impaired cerebral blood flow reserve group.And in the trial group the brain structural reserve was positively correlated to the infarct volume and the NIHSS score,and the rank correlation coefficient was 0.219 and 0.238 respectively (P ＜ 0.05).The prognosis of cerebral blood flow reserve in the normal group was better than the impaired group (x2 =4.155,P ＜ 0.05),for example,the proportion of patients with good prognosis and normal CBFV was 18.84％ (13/69),the proportion of patients with good prognosis but decreased CBFV was 81.16％ (56/69),the proportion of patients with poor prognosis but normal CBFV was 2.70％ (1/37),the proportion of patients with poor prognosis and decreased CBFV was 97.30％ (36/37).The proportion of patients with type Ⅰ and type Ⅲ of the brain structure reserve was 37.68％ (26/69) and 5.80％ (4/69) respectively,whose prognosis was better (x2 =8.456,P ＜ 0.05) than patients with type Ⅱ and type Ⅳ,whose proportion was 43.48％ (30/69) and 13.04％ (9/69).Multivariate Logistic regression analysis showed that NIHSS score,age,and brain structural reserve were risk factors for poor prognosis in the trial group.Normal cerebral blood flow reserve was a protective factor for good prognosis.Conclusions CVR in patients with acute atherosclerotic cerebral infarction is significantly reduced.CVR can be used as an index to evaluate the prognosis of patients who were followed-up for three months.