BACKGROUND:Calcium citrate has a better solubility than calcium phosphate, calcium sulfate, and other calcium biomaterials. The synthetic calcium citrate has a good denseness, and stably releases calcium ions at a high efficiency during the degradation. Consequently, it may be more suitable for the fil ing of fracture defects, providing needed calcium ions for early fracture healing.
OBJECTIVE:To prepare calcium citrate biomaterials with a novel formulation based on the natural bio-mineralized oyster shel s and citric acid so as to expect to get a good application in fracture healing repair.
METHODS:Crushing, grinding, and chemical reaction methods were used for refinement. Particle size analyzer, X-ray diffraction, scanning electron microscope, and Fourier transform infrared spectroscopy were adopted for analysis of the size distribution, composition, mineral phases, and micro-morphology. Biological characteristics were evaluated through a simulated body fluid experiment.
RESULTS AND CONCLUSION:Oyster shel powder was reacted with saturated citric acid to produce the calcium citrate material that had uniform crystal structure and compact bonding among crystal bodies, and exhibited a certain mechanical ability. The calcium citrate material had a good crystal structure that was conductive to prolong the degradation time. The calcium citrate released calcium ions slowly, and did not produce dramatic changes in the pH value (7.20-7.46) of the surrounding in the dissolution process. With the gradual degradation of calcium citrate materials, Ca2+concentration in solution increased gradual y and stably, and ultimately achieved an appropriate concentration of 7 mmol/L, suitable for osteoblast proliferation and differentiation. Calcium citrate prepared using natural oyster shel has good biological properties, and exhibits a natural superiority to artificial bone materials.

China ; Foundations ; Preventive Medicine ; economics ; Research Support as Topic ; statistics & numerical data

Objective To study environmental safety reference standard values of commonly encountered pathogenic microorganisms, and provide the basis for environmental risk assessment of a high-level biosafety laboratory (BSL).Methods Using human exposure dose calculation methods as the basic principle in combination with microbial hazard evaluation methods , an infectious dose calculation model of human exposure to pathogenic microorganisms by breathing was proposed . Based on research on characteristics of commonly encountered pathogenic microorganisms at home and abroad , the environ-mental safety threshold reference standard and environmental risk protection zone for such microorganisms were obtained . Results An the environmental risk assessment standard calculation method for commonly encountered highly pathogenic microorganisms was proposed , and the minimum infectious dose and environmental safety threshold of seven commonly encountered pathogenic microorganisms were obtained , including SARS coronavirus , highly pathogenic avian influenza virus, Mycobacterium tuberculosis, Bacillus anthracis, Yersinia pestis,Francisella tularensis, and epidemic hemorrhagic fever virus.Conclusion The results can be used as a reference standard of infectious aerosols in a high-level BSL for environ-mental risk assessment , laboratory risk control and emergency management .

AIM: To explore the effect of hypertonic sugar eyes drops with local to the oxygen therapy for severe corneal edema after cataract surgery.METHODS: Totally 68 patients (68 eyes) with severe corneal edema after phacoemulsification were selected from January 2014 to December 2015 in our hospital, who were aged 53-80(64.45±4.24), including 30 males and 38 females.According to different treatment, they were divided into treatment group (34 cases) given conventional therapy + hypertonic sugar eyes drops with local to the oxygen therapy, the control group (34 cases) given conventional treatment + hypertonic sugar eyes drops.Visual acuity and corneal situation were observed during the treatment.RESULTS: Corneal edema fade time of treatment group was 11.62+0.53d, that of control group was 15.23±0.62d, the difference between the two groups was significant (P<0.05).Preoperative best corrected visual acuity (BCVA) and corneal endothelial cell count had no significantly difference between the two groups while the corneal endothelial cell count were significantly different compared with postoperative of the two groups (P<0.05).At 7d after treatment, the BCVA and corneal endothelial cell count had significantly difference between the two groups (P<0.05), which did not have significant difference at 1mo after treatment (P>0.05).The effective rates of the two groups was significantly different (P<0.05).CONCLUSION: Hypertonic sugar eyes drops with local to the oxygen therapy is effective for severe corneal edema after cataract surgery.

Objective To identify serum biomarkers for rheumatoid arthritis(RA)by protein finger- print pattern. Methods One hundred and forty-one serum samples of 90 RA patients, 20 systemic lupus ery- thematosus(SLE)patients, and 31 healthy individuals were randomly divided into training set(n=93, 60 RA patients, 13 SLE patients and 20 healthy individuals)and test set(n=48, 30 RA patients, 7 SLE patients and 11 healthy individuals). They were detected by surface enhanced laser desorption/ionization-time of flight- mass spectrometry(SELDI-TOF-MS). The protein fingerprint pattern obtained from SELDI-TOF was trained by a multi-layer artificial neural network(ANN)to establish a diagnostic model. Results The detective mod- el obtained by ANN was used to detect the 48 unknown serum samples. The sensitivity and specificity for RA detection was 90% and 90.9% respectively. Conclusion In comparison with traditional methods, SELDI- TOF-MS could identify new serum biomarkers in RA. Combined with ANN, it provides high sensitivity and specificity for RA diagnosis.

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.

After 28 foreign species of AM fungi were inoculated in sterilized soil, the effects of the AM mycorrhizal colonization and the medicine quality of Paris polyphylla var. yunnanensis were observed by combination of inoculation test in pot at room temperature and instrumental analysis. The results showed that, compared with control group (CK), the inoculation of foreign AM fungi in the soil influenced the spore density, mycorrhizal infection rate, and colonization intensity of AM fungi in root system of P. polyphylla var. yunnanensis. The inoculation of foreign AM fungi enhanced the mycorrhiza viability of P. polyphylla var. yunnanensis by increasing the activity of succinic dehydrogenase (SDH) and alkaline phosphatase (ALP) in intraradical hyphae. The content of single steroid saponin in rhizome of P. polyphylla var. yunnanensis showed variation after P. polyphylla var. yunnanensis was inoculated by different foreign species of AM fungi, which was beneficial for increasing the medicine quality; however, the kinds of steroid saponin showed no difference. In a degree, there was a selectivity of symbiosis between P. polyphylla var. yunnanensis and foreign AM fungi. And we found that the Claroideoglomus claroideum and Racocetra coralloidea were best foreign AM fungi species for cultivating P. polyphylla var. yunnanensis under field condition.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Fungi ; classification ; growth & development ; Liliaceae ; chemistry ; growth & development ; microbiology ; Mycorrhizae ; classification ; growth & development ; Plant Roots ; chemistry ; growth & development ; microbiology ; Quality Control

OBJECTIVE:To establish the method for the determination of mildronate in human plasma and urine,and to study the pharmacokinetic characteristics in healthy volunteers. METHODS:After precipitating plasma and urine sample,LC-MS/MS method was adopted. Dikma Diamonsil C18 column was used with mobile phase consisted of methanol-water(containing 0.2% for-mic acid,0.3% ammonium acetate)(31∶69,V/V)at the flow rate of 0.6 ml/min. ESI was adopted in MRM mode,by using nega-tive ion. The ion for quantitative analysis were m/z 147.10→58.20 (mildronate) and m/z 152.00→110.10 (internal standard,acet-aminophen). The pharmacokinetic parameters of mildronate with single administration and multiple administration were calculated by using DAS 2.1 software and compared. RESULTS:The linear range of mildronate in plasma were 0.02-20 ng/ml(r=0.999 3) and in urine were 0.05-40 ng/ml(r=0.998 2). The lowest limits of quantitation were 0.02 and 0.05 ng/ml. Precision and recovery met the requirements of biological specimen determination,and endogenous impurities hadn’t effect on the determination. The main pharmacokinetics parameters of low-dose,medium-dose and low-dose(250,500,750 mg)of mildronate in plasma with single ad-ministration were as follows:t1/2 were(3.39±0.81),(5.52±0.57)and(5.32±0.96)h;tmax were(0.80±0.45),(1.38±0.43)and (1.10±0.36)h;cmax were(4.17±1.46),(8.08±1.04)and(15.04±1.86)ng/ml;AUC0-36 h were(24.55±5.81),(45.50±7.07)and (85.60 ± 13.09)ng·h/ml. In the dose range,cmax,AUC0-36 h h had a linear relationship with dose (R2 were 0.974 5 and 0.968 3). The main pharmacokinetic parameters of low-dose of mildronate with multiple administration after keeping stable were as follows:cmin was(0.28 ± 0.10)ng/ml;AUCs was(38.78 ± 4.18)ng·h/ml;cs was(1.62 ± 0.17)ng/ml;DF was(3.81 ± 1.14);t1/2 was(6.17 ± 1.46)h;tmax was(1.20 ± 0.33)h;cmax was(6.46 ± 1.96)ng/ml;AUC0-36 h was(40.33 ± 4.65)ng·h/ml;accumulation factor of cmax and AUC were(1.73±0.90)and(1.64±0.40). Compared with single administration,t1/2,cmax and AUC of mildronate with multiple admin-istration after keeping stable all changed,and tmax had no signifi-cant difference. After single administration,26 h accumulative excretion rate of those groups were (0.004 009 ± 0.001 1)%, (0.004 026±0.001 01)% and(0.003 858±0.000 68)% respec-tively. CONCLUSIONS:Established method is sensitive,accurate and specific,and suitable for the determination of mildronate concentration in human plasma and urine and pharmacokinetics study. Mildronate capsule shows certain accumulation effect in healthy volunteers,and linear pharmacokinetic characteristics.

Objective To detect the expressions of CD19 and CD20 of B-lymphocytes (CD19+B, CD20+B) and natural killer (NK) cells in the peripheral blood of epileptic children and explore their significances. Methods Four hundred and fifty-eight epileptic children, admitted to our hospital from January 2008 to December 2010, were chosen as patient group; another 52 healthy subjects were chosen as controls. The expressions of CD19+B and CD20+B and NK cells were detected by flow cytometry; their results were compared. Ninety-two epileptic children were treated with intravenous immunoglobulin (IVIG) and the effect of IVIG therapy was studied. Results The CD19+B level ([22.35±6.54]％) and CD20+B level ([21.50±8.41]％) in the epileptic children were obviously higher than those in the healthy controls ([16.86±4.02)％,[16.13±4.19]％, P＜0.05); and the level of NK cells ([9.1 1±4.90]％) in the epileptic children was significantly decreased as compared with that in the healthy controls ([14.72±4.15]％, P＜0.05). The CD19+B level ([18.26±5.03]％) and CD20+B level ([16.74±5.12]％) 6months after MG treatment were decreased significantly as compared with those before treatment ([22.74±6.25]％,[21.61±8.03]％, P＜0.05]; while the level of NK cells ([14.65±4.58]％) 6 months after IVIG treatment was increased significantly as compared with that before treatment ([9.07±4.76]％,P＜0.05). Among the 92 patients treated with IVIG, 70 enjoyed good results and 22 had non-effective resutls; The changesofCD19+Blevel ([7.99±5.34]％) and CD20+B ([8.21±5.21]％) before and after treatment in effectively treated patients by IVIG were significantly different as compared with those in ineffectively treated patients by MG ([3.78±2.76]％,[3.66±2.48]％, P＜0.05); however, the changes of level of NK cells before and after treatment showed no significant difference between effectively treated patients and ineffectively treated patients ([5.28±4.55]％,[4.53％±4.43]％, P＞0.05). Conclusion Dysfunctions of B-lymphocytes and NK cells exist in epileptic children, and MG treatment shows good effect on immune dysfunction of them. The levels of CD19-B and CD20-B can be used as monitoring indicators in the IVIG treatment of epileptic children.

Optimization of the fermentation condition for human apolipoproteinA-I expression in recombinant Escherichia coli was investigated. The recombinant plasmid pBV220-ApoA-I was transformed respectively into different E.coli hosts such as JM109, BL21(DE3),DH5?, BMH7118,and TG1. The best host E.coli was DH5? in which the recombinant ApoA-I expression percentage was 21.2% corresponding to that in BL21(DE3) in flask shaker cultivation,while the ApoA-I expressed percentage in E.coli TG1 was 11%.Fed-batch cultivation was performed in FMG-5L fermentor,the optimum fermentation cultivation conditions were as following :optimum pH value was 7.0 in growth phase and 7.4 in the expression phase. The initial glucose concentration in batch phase was 3 g?L -1.The optimum C/N ratio was 2∶1.The recombinant ApoA-I reached about 40% of the total protein, and concentration of ApoA-I was 2.86 g?L -1.