A rapid method for the simultaneous determination of berberine (BBR), matrine (MT) and oxymatrine (OMT) by nonaqueous capillary electrophoresis (NACE) was developed. Optimum separation of the analytes was obtained on a 50cm×50μm i.d. fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate, 7.0% acetic acid and 10% acetonitrile at 25kV and 20℃. The relative standard deviations (R.S.D.) of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine, 0.11%-0.60% and 0.74%-1.63% for matrine, 0.15% and 0.45% for oxymatrine, respectively. Detection limits of berberine, matrine and oxymtrine were 0.18μg/mL, 4.08μg/mL and 4.16μg/mL, respectively. In the tested concentration range, good linear relationships (0.9992 for berberine, 0.9988 for matrine and 0.9988 for oxymatrine) were observed. The linear calibration ranges were 0.45-360.0μg/mL for berberine, 8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine. This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen) and Cortex phellodendri chinensis (Huangbai) and their medicinal preparations.

Objective To investigate the correlation between the degree of pathogenetic condition and cogni-tive impairment in young OSAHS patients. Methods Sixty-three patients (18～44 years old) were divided into 3 groups according to the assessment criteria regarding the degrees of pathogenetic conditions of hypoxemia and its persistence time: mild group(the lowest SaO2 ≥ 85 %, n = 20 ), moderate group (the lowest SaO2 : 65 ～ 84 %, n =24), and severe group (the lowest SAO2<65%, n= 19). Twenty-five healthy young adults were selected as the control group. All the subjects were tested with event-related potential, namely P300, the polysomnogram and the mini-mental state examination scale(MMSE). Results P300 latency in patients of three OSAHS groups was sig-nificantly prolonged as 326.1±12.7, 346.9±19.1, and 34.9±18.3 ms, as compared with those of control group: 311.9±18.3 ms (P< 0.05, respectively). In severe group, P300 latency was markedly increased compared with mild and moderate groups(P<0.01, P<0.05, respectively). No obvious differences were observed between mild and moderate groups(P=0. 095). The difference of hypoxemia lasting time could influence the P300 latency, such as when the hypoxemia lasting time was 4～60 seconds, the latency was 338. 12±13.7 ms, and when the hypoxemia lasting time increased to 60 ～ 140 s, the latency was prolonged to 354.74±16.7 ms(P = 0. 031 ). There was no difference among all groups in the P300 amplitudes. The MMSE scores for all patients were within normal limits andthere were differences between serve and control group(RA -RB=9. 91, P= 0. 003). Conclusion Among patients with OS-AHS, the impairment of cognitive function is con'anon. The degrees of pathogenetic conditions of hypoxemia and its lasting time may be used to evaluate the impairment degree of cognitive function in patients with OSAHS.

A growing number of children and adolescents are being diagnosed as Chiari malformation type Ⅰ (CM-Ⅰ) for behavioral disorders,developmental delay,seizures,or abnormal orpharyngeal function.The aim of this study was to compare the clinical characteristics,imaging findings and surgical outcomes of CM-Ⅰ in pediatric and adult patients.Between January 2014 and June 2017,84 patients with CM-Ⅰ underwent surgical treatment in our department.We divided the patients into two groups:pediatric group (n=1 1,age ＜18 years)and adult group (n=73,age ≥18 years).Data on clinical characteristics,imaging findings,surgical outcomes,and prognosis were retrospectively reviewed and compared between these two groups.For clinical presentation,scoliosis (36.4％) and developmental delay (36.4％) were more common in pediatric patients,whereas,sensory disturbance (58.9％) and motor weakness (41.1％) were more common in adult patients.Imaging findings showed that the incidence of hydrocephalus and craniovertebral junctional abnormalities was significantly higher in pediatric group than in adult group (P＜0.05).Compared to adult group,pediatric group showed a better improvement or resolution of syrinx and tonsillar herniation after surgical treatments (P＜0.05).The total Chicago Chiari Outcome Scale (CCOS) score in pediatric patients at the last follow-up was significantly higher than that in adult patients (P=0.002).In conclusion,the clinical characteristics and imaging findings appeared to be different in pediatric and adult patients with CM-Ⅰ.The surgical outcomes of pediatric patients were shown to be significantly better than those of adult patients.

Exhaled breath condensate(EBC) is a new method for diagnosis and evaluation of lung disease,EBC compositions form to the fluid lining airway surfaces.EBC examination is a safe,non-invasive,easy and simple diagnostic method that is suitable for applicable in infants and childhood.Changes of EBC composition occurs in the airway inflammation or injury.There are changes in the pH value,inflammatory mediators,cytokines and other composition in EBC of childhood asthma.Analysis of EBC composition can contribute to diagnosis and severity assessment of childhood asthma.

Objective To investigate the alterations of connexin 43 (Cx43) expression and its distribution at different stages of myocardial infarction (MI) in rats after transplantation of allogenic mesenchymal stem cells (MSCs).Methods Wistar rats were ligated on the left anterior descending coronary artery to make MI models.They were injected with allogenic MSCs,which were induced by 5-aza and labelled by DAPI,during the second operation after 7 days of MI.In subgroups,MSCs were detected by fluorescence microscope.Cx43 expression and GJ distribu-tion were examined by immunohistochemistry after 4,8 or 12 weeks respectively.Results MSCs differentiated into cardiac muscle cell-like cells which were capable of pulsing spontaneously,expressing cTnT and forming myofilament in vitro.Transplanted MSCs can survive in MI host and upregulate Cx43 expression and normalize Cx43 distribution at ischemic zones after 4,8 and 12w.No change of Cx43 was seen at infarcted zones.Conclusion MSCs have the plasticity of differentiating into cardiac muscle cell-like cells which can continuously upregulate Cx43 expression and normalize Cx43 distribution at ischemic zones after 4,8 and 12w.

BACKGROUND: Stem cell transplantation in repairing infarct myocardium and in improving cardiac function has been widely accepted. However, whether transplanted cells and host cells formed an effective electricity and mechanical couple, whether a relevant independent electrical system with contractile function formed or whether severe malignant ventricular arrhythmia formed, are still unclear.OBJECTIVE: To investigate electrophysiological abnormaltiy and left ventricular remodeling in rats with myocardial infarction following allogenic bone marrow mesenchymal stem cell (BMSC) transplantation.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Experimental Center, Guangxi Medical University from December 2005 to October 2008.MATERIALS: A total of 120 healthy Wistar rats were equally randomized into normal control, sham operation, saline control and cell transplantation groups. Healthy Wister rats aged 1 month were selected to harvest bone marrow.METHODS: At the third passage, rat BMSCs were collected and treated with 5-aza, and differentiated into cerdiomyocytes.BMSCs were labeled with DAPI at 2 hours before transplantation. In the saline control and cell transplantation groups, rat models of myocardial infarction were established by ligating the left anterior descending coronary artery. In the sham operation group, the coronary artery was not ligated, but only braid. At 7 days following ligation, BMSCs in the cell transplantation group at 2×10-1/L were infused into the edge and center of myocardial infarct region by multipoint injection. Rats in the other three groups were subjected to an equal volume of saline.MAIN OUTCOME MEASURES: Electrocardiogram and cardiac electrophysiology were performed. Ultrasonic cardiography was used to detect left ventricular function. Infarct size was determined. DAPl-labeled donor cell migration and distribution was observed with a fluorescence microscope.RESULTS: BMSCs could differentiate into cardiacmuscle cell-like cells which were capable of pulsing spontaneously, expressing cardiactoponin T and forming myofilament in vitro. Compared with the saline control group, PR interval, QRS duration and ventdcular effective refractory period shortened, ventricular fibrillation threshold increased at 4, 8 and 12 weeks (P < 0.05); left ventricular internal diameter at end-systole reduced, and left ventricular ejection fraction and shortening traction was significantly increased (P< 0.05). At 8 and 12 weeks, infarct size was significantly smaller (P < 0.05). At 4 weeks, DAPl-labeled BMSCs could be seen under the fluorescence microscope, and still could he detected at 12 weeks. However, the fluorescence became weak with prolonged time.CONCLUSION: BMSCs have the plasticity of differentiating into cardiac muscle cell-like cells, which can modulate theelectrophysiological abnormality and left ventricular remodeling following myocardial infarction.

Objective To investigate the antimicrobial resistance of clinical isolates from a hospital in Chongqing during one year according to CHINET project.Methods Disc diffusion test (K-B method) was employed to study the antimicrobial resistance. WHONET5 was used for data analysis.Results In one year period from 2004 to 2005,690 non-duplicate isolates were collect- ed.Enterobacter isolates showed the lowest resistance rate to imipenem and meropenem.About 37.5% of E.coli and 31.4% of K.pneumoniae isolates produced ESBLs,respectively.All ESBLs-producing strains were susceptible to imipenem and mer- openem.About 37.2%,39.4% and 48.9% of P.aeruginosa isolates were resistant to imipenem,meropenem and ceftazidime, respectively.Pandrug-resistant (PDR) P.aeruginosa was isolated from our hospital.All strains of A.baumannii were sus- ceptible to imipenem and meropenem.About 37.7% of A.baumannii were resistant to cefoperazone-sulbactam.Twenty-nine strains showed the same resistant pattern among non-susceptible strains of A.baumannii,mainly derived from 2 clones by PFGE analysis.Conclusions The surveillance results suggest that prevalent strain resistant to cefoperazone-sulbactam may pres- ent in some ICUs.Resistance rate to cefoperazone-sulbactam increased significantly.

The association rate constant and dissociation rate constant are important parameters of the drug-cyclodextrin supermolecule systems, which determine the dissociation of drugs from the complex and the further in vivo absorption of drugs. However, the current studies of drug-cyclodextrin interactions mostly focus on the thermodynamic parameter of equilibrium constants (K). In this paper, a method based on quantitative high performance affinity chromatography coupled with mass spectrometry was developed to determine the apparent dissociation rate constant (k(off,app)) of drug-cyclodextrin supermolecule systems. This method was employed to measure the k(off,app) of meloxicam and acetaminophen. Firstly, chromatographic peaks of drugs and non-retained solute (uracil) on β-cyclodextrin column at different flow rates were acquired, and the retention time and variance values were obtained via the fitting the peaks. Then, the plate heights of drugs (H(R)) and uracil (H(M,C)) were calculated. The plate height of theoretical non-retained solute (H(M,T)) was calculated based on the differences of diffusion coefficient and the stagnant mobile phase mass transfer between drugs and uracil. Finally, the k(off,app) was calculated from the slope of the regression equation between (H(R)-H(M,T)) and uk/(1+k)2, (0.13 ± 0.00) s(-1) and (4.83 ± 0.10) s(-1) for meloxicam and acetaminophen (control drug), respectively. In addition, the apparent association rate constant (k(on,app)) was also calculated through the product of K (12.53 L x mol(-1)) and k(off,app). In summary, it has been proved that the method established in our study was simple, efficiently fast and reproducible for investigation on the kinetics of drug-cyclodextrin interactions.
Acetaminophen ; chemistry ; Chromatography, Affinity ; Drug Interactions ; Kinetics ; Mass Spectrometry ; Thermodynamics ; Thiazines ; chemistry ; Thiazoles ; chemistry ; beta-Cyclodextrins ; chemistry

This study was aimed to evaluate whether human mesenchymal stem cell (MSC) and the Stro-1 positive subgroup have inducing immune tolerance effect in mouse skin graft model. Human MSC were isolated and cultured from bone marrow-derived mononuclear cells of healthy adults, and Stro-1 positive cells were sorted out. Female C57BL/6 mice and female BALB/c mice were respectively used as donors and recipients in skin allogenic graft model. The recipients were divided randomly into 4 groups: (1) Stro-1(+) MSC group: 2×10(6) Stro-1(+) MSC were injected into the irradiated recipient mice before skin graft. (2) MSC group: 2×10(6) MSC were injected into the irradiated recipient mice before skin graft. (3) Irradiation control group: the recipient mice were just irradiated before skin allogenic graft. (4) Congenic control group: the irradiated BALB/c mice received the skin from the congenic mice. The survival time and pathologic changes of skin grafts were observed by macro- and microscopy with HE staining. The transforming growth factor β1 (TGF-β1) concentration in plasma of recipient mice was measured by ELISA before and after grafting. The results indicated that the survival time of skin grafts in the MSC group was (12.13 ± 3.34) d, which was not notably longer than the irradiation control group (11.38 ± 1.01) d. The survival time of skin grafts was significantly prolonged in the Stro-1(+) MSC group (30.68 ± 5.89) d, as compared with the irradiation control group and the MSC group, respectively; the pathologic examination of skin grafts showed a clear structure. After grafting, the TGF-β1 concentration in plasma of recipient mice was almost the same as before grafting in the irradiation control group and the congenic control group, but it significantly increased in the MSC group and the Stro-1(+) MSC group. It is concluded that the Stro-1(+) MSC induce greater immune tolerance than the unsorted MSC, and significantly prolong the survival time of skin grafts in vivo, while TGF-β1 does not contribute to the immune tolerance.
Animals ; Female ; Graft Survival ; immunology ; Humans ; Immune Tolerance ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Skin Transplantation ; immunology ; methods ; Transforming Growth Factor beta1 ; blood ; Transplantation, Homologous

OBJECTIVE To investigate the clinical significance of human papillomavirus in paraffin-embedded cervical cancer and precancerous lesion tissue by gene clip technology.METHODS 153 Patients with paraffin-embedded examples.DNA was extracted from paraffin-embedded tissues and amplified by polymerase chain reaction(PCR).RESULTS The positive rate of high-risk HPV of inflammation was 8.33%,CINⅠ45.83%,CINⅡ/CINⅢ 87.50% and invasive cancer 92.21%.The HPV infection rate of squamous cell carcinoma was 94.12%.The HPV infection rate of adenocarcinoma was 88.46%.Among all the patients with cervical cancer and CIN,the infection rate of HPV16,the most genotype,was 88.98%.The infection rate of HPV18,the second most subtype,was 33.06%.In addition,the minority were infected HPV52、33、59、68.Among 48 cases of cervical squamous cell carcinoma,the infection rate of HPV16,HPV18 was 93.73% and 27.08% respectively.Among 23 cases of cervical adenocarcinoma,the infection rate of HPV16,HPV18 was 82.61% and 52.17% respectively.On the other hand,all the patients with cervicitis were HPV single infection.The HPV multiple infection rate of CINⅠ,CINⅡ/CINⅢ,cervical cancer was 20.00%,28.57%,36.62% respectively.CONCLUSIONS Gene chip technology can detect multiple HPV genotypes in paraffin-embedded tissues with high sensitivity and specificity,which is useful in the pathogenesis and prevention of cervical cancer.