Objective To investigate the infection sites,the offending species,diagnosis and prognosis of deep fungal infections (DFI) in patients with systemic lupus erythcmatosus (SLE).Methods Fifty-one patients with fungal infections in 1466 SLE patients admitted to Peking Union Medical College (PUMC) Hospital from 2000 to 2006 were reviewed retrospectively.Results Candida albicans was ranked the first pathogen,followed by ncoformans and Aspergillus species.The infection sites were lungs,cerebral meninges and blood in the order of prevalence.The overall mortality was 20%(10/51).Aspergillasis carried the highest mortality which couht be as high as 80%.Hypoproteinemia,multiple focus of fungal infections,Aspergillasis and fungemia might he the independent risk factor for mortality.Conclusion Candida albicans is the most frequent species of fungal infections in SLE patients.Iungs are the most prevalent location of infection.Earlier diagnosis is important.Special attention should be paid to aspergillasis.

Objective To explore the effect of chromofungin (CHR), a chromogranin A (CGA) derived peptide CGA47-66, on hyper-permeability of blood brain barrier in septic mice. Methods 120 healthy male C57BL/6 mice were randomly divided into groups, with 12 mice in each group. Seventy-two mice were used for dynamic observation of the contents of water and Evan blue (EB) in brain tissue after being treated with lipopolysaccharide (LPS). Another 48 mice were divided into normal saline control group (NS group), LPS induced sepsis model group (LPS group), low-dose CHR pretreatment group (CL+LPS group), and high-dose CHR pretreatment group (CH+LPS group). The septic model was reproduced by intraperitoneal injection of 10 mg/kg LPS 0.1 mL, and the mice in NS group was given equal volume of normal saline. The mice in CL+LPS group and CH+LPS group were intraperitoneally injected with 15.5 μg/kg and 77.5 μg/kg CHR 10 minutes before LPS injection. Six hours after LPS injection, 4 mL/kg of 2% EB was injected via caudal vein, the contents of water and EB in brain tissue were determined, and EB immune fluorescence in brain tissue was determined to assess the changes in permeability of blood brain barrier. Brain pathology was observed with hematoxylin and eosin (HE) staining. Results With the extension of time after LPS injection, the contents of water and EB in brain tissue were gradually increased, and the time of difference with statistical significance appeared earlier when compared with that of control group in the contents of water than that in EB contents (3 hours and 6 hours, respectively). The contents of water and EB in brain tissue in LPS group were significantly increased as compared with NS group [water content: (79.77±0.62)% vs. (78.28±0.44)%, P < 0.01; EB content (μg/g): 13.87±4.50 vs. 7.13±1.76, P < 0.05]. CHR pretreatment with either of two dosages could reverse the increase in water and EB contents in brain tissue induced by LPS, and the effect was more significant in CH+LPS group [water content: (78.15±0.73)% vs. (79.77±0.62)%, EB (μg/g): 7.09±2.59 vs. 13.87±4.50, both P < 0.05]. It was shown by EB fluorescence observation that the fluorescence signal displayed only in the meninges in NS group, and EB fluorescence was widely distributed in brain parenchyma in LPS group, indicating that the EB leakage in LPS group was more marked than that of NS group. In CHR pretreatment groups, EB fluorescence was decreased in brain parenchyma, indicating that EB leakage was significantly less marked, while it was more obvious in high dose CHR group. It was shown by HE staining that cerebral blood vessel structure was intact in NS group, and the gap around blood vessel was not significant increased. On the other hand, brain structure in LPS group appeared loose, with widening of small perivascular spaces and obvious edema. Brain edema in CHR pretreatment groups was improved as compared with that of the LPS group, and it was more apparent in high dose CHR group. Conclusions LPS induced change in blood brain barrier permeability in mice in a time-dependent manner. Exogenous CGA derived peptides CHR can inhibit LPS induced hyper-permeability of blood brain barrier in septic mice, thus reduces brain edema, protects the brain tissue, and the effect is more obvious with a high dose of CHR (77.5 μg/kg).

The paper introduces the research idea, design and realization of the distributed Naive Bayesian intelligent diagnosis sys-tem based on Hadoop, makes optimization and improvement according to its application in Traditional Chinese Medicine ( TCM) Hospital of Guangdong Province, including algorithm design improvement and enhancement of accuracy, extensibility and security of the system.

Objective To investigate the pharmacodynamics of different local anesthetics administered intrathecally for elderly patients undergoing transurethral resection of the prostate (TURP). Methods Ninety ASA Ⅰ - Ⅲ elderly patients, aged 69-82 yr, with body mass index less than 30 kg/m2 , undergoing TURP under combined spinal-epidural anesthesia, were randomly divided into 3 groups ( n = 30 each): levobupivacaine group (group L), ropivacaine group (group R) and bupivacaine group (group B). Group L, R and B received intrathecai (IT) 0.5 % levobupivacaine, 0.5 % ropivacaine and 0.5 % bupivacaine respectively. The initial dose was 7,10 and 6 mg in group L, R and B respectively. The ratio of two successive doses was 0.9. If the upper sensory block reached T10 within the 20 min after IT injection, the IT analgesia was considered to be effective. The median effective dose (EDs0) and 95 % confidence interval (95 % CI) were calculated by Dixon. Results The ED50 and 95% CI of levobupivacaine, ropivacaine and bupivacaine were 6.781 (95% CI 6.561-7.024) mg, 9.135 (95%CI8.670-9.616) mg and 5.170 (95% CI 5.012-5.333) ng respectively. The relative potency ratio between levobupivacaine, ropivacaine and bupivacaine is 0.76∶0.57∶1.00. ConclusionThe relative potency ratio be tween levobupivacaine, ropivacaine and bupivacaine is 0.76∶0.57∶1.00.

The ORFK8.1 of Kaposi's sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogalactopyranoside (IPTG). The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis (SDS-PAGE). SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant 0RFK8.1 protein. The optimal condition of the recombinant ORFK8.1 ELISA assay was confirmed: the concentration of antigen was 5 ug/mL, the dilution of serum was 1:200. We used the ELISA method to investigate the recombinant ORF K8.1 protein's specificity, the data showed that the specificity of ORF K8.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFK8.1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.

Objective To evaluate the effect of different preparation processes on preparation of the glial cell line-derived neurotrophic factor(GDNF)loaded microspheres and observe the biological activity of GDNF.Methods With polylactide-co-glycolide(PLGA)as the coating material,the GDNF-loaded microspheres were prepared by using double emulsion(W1/O/W2).Two-factor factorial design variance analysis was done to analyze the effects of the composition proportion of lactic acid(LA)and glycolic acid(GA)in PLGA and the stirring speed of multiple emulsion on particle size,entrapment efficiency,burst release and in vitro release characteristics of the GDNF-loaded microspheres.PC-12 bioassay was employed to detect the biological activity of the released GDNF so as to determine the optimal preparation process.Results The composition proportion of PLGA could affect the microspheres'burst release(P ＜ 0.05),with no effect on particle size and entrapment efficiency.with the higher.With higher proportion of GA,the release speed of GDNF in the microspheres was increased.When the stirring speed of multiple emulsion was increased from 1 000 r/min to 3 000 r/min,the particle size of the microspheres was decrease significantly(P ＜ 0.01),the burst release was increased markedly(P ＜ 0.01)and the in vitro release rate was accelerated.The activity of GDNF in the microspheres could last for about 20 days at 37℃,which was 10 days longer than that of single GDNF.Conclusions Double emulsioncan prepare the GDNF-loaded microspheres with high entrapment efficiency and suitable in vitro release time.In the meantime,the microspheres can extend the validity of GDNF.

Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS),primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Hart people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).

Objective To investigate how to diagnosis traumatic subdural effusion (TSE) and differentiate it with senile encephalatrophy in aged people by CT scan. Methods Fifty-two aged patients with TSE and 40 aged patients with encephalatrophy, admitted to our hospital from March 2008 to March 2010, were chosen in the study. Their clinical data and CT manifestations were retrospectively analyzed and compared. Results Fifty-two patients with TSE had 91 sites of the lesion, including 31 lesions located in temporal-frontal lobe and 30 lesions in frontal lobe. Among the 52 patients, 34 were also noted as having encephalatrophy. First-time CT demonstrated 32 patients with TSE, and the second-time CT on the left patients showed 20 having TSE too. Among the 52 patients, increasing amount of fluid in a short term was noted in 32 patients; local widened gyri with a flattened surface and narrowed sulci was presented in 30; "compressed arachnoid sign" was found in 9. CT results demonstrated that no above-mentioned manifestations were presented in patients with encephalatrophy.Conclusion For aged person, TSE is usually located in the temporal and frontal lobe, and should be differentiated with senile encephalatrophy. The key-points for differentiation include the location and the extent of changes of extra-cerebral space, the space-occupying signs as local flattened gyri and narrowed sulci, the appearing of "compressed arachnoid sign" and the changes of effusion in a short-term review.

The gene of mutated TNF-?D4 gene was amplified by overlap PCR and cloned into the prokaryotic expressive vector pBV220.TNF-?D4 contains two changes:substitutions of Pro8Arg,Ser9Lys,Asp10Arg,Ile157Phe,Leu29Ser,Arg31Val and a deletion of the N terminal four amino acids.The recombinant vector pBV220-TNF-?D4 was transformated into E.coli strain DH5?,and the high expression strain was obtained by screening monoclones.The level of expression was about 45% of total cell protein.After purification,the purity of fusion protein was above 90% by HPLC and relative ability was 8 ?107.TNF-?D4 was modificated by mPEG-ButyrALD。After purification,the purity of mPEG-TNF-?D4 was above 85% and relative ability was 8.6?107.The in vivo systemic toxicity of mPEG-TNF-?D4,which is indicated by LD50,is lower than that of rhTNF-?.These results strongly supported for the further study and exploitation of TNF-antitumor drug.

Three homoisoflavonoids, 6-aldehydo-3-ophiopogonanone A, methyl ophiopogonanone A and ophiopogonanone A from Ophiopogon japonicus were analyzed simultaneously by HPLC with acetonitrile-water containing 0.5% H3 PO4 (58:42) as the mobile phase, and the detection wavelength was set at 296 nm (0-14 min) and 275 nm (14-22 min). The mean recoveries of three homoisoflavonoids were 99.41%-99.86% (RSD 0.82%-1.05%). The linear response ranges of. 6-aldehydo-3-ophiopogonanone A, methyl ophiopogonanone A and ophiopogonanone A were 0.165-0.990 microg (r = 0.999 9), 0.153-0.918 microg (r = 0.999 9), and 0.270-1.620 microg (r = 0.999 9), respectively. This method was certified to be accurate and reliable and can be used for quality control of O. japonicus.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Isoflavones ; analysis ; Ophiopogon ; chemistry