Abstract?AlM:To investigate the effects of the MUC5AC levels and ocular function of the patients with glaucoma and cataract with combined surgery.? METHODS: Twenty - eight patients treated with glaucoma and cataract combined surgery were chosen as the observation group from December 2011 to June 2014 in our hospital, and other 28 cases of glaucoma and cataract did not undergo surgical treatment were selected as the control group, 30 healthy subjects were as healthy control group. the MUC5AC level and ocular surface score of the three subjects before surgery 1d, after 3 and 6mo were compared.?RESULTS: The NUC5AC of the two groups of patients was significantly lower than that of the healthy control group before surgery (P<0. 05), the ocular function score was significantly higher than the healthy control group ( P<0. 05). After 1mo, the MUC5AC of the observation group were significantly lower than that of before surgery ( P<0. 05), after 3mo MUC5AC content gradually increased to preoperative levels, after 6mo the MUC5AC were significantly higher than before surgery (P<0. 05). After 1mo, ocular function scores were significantly higher than the preoperative ( P< 0. 05 ), while after 3mo, ocular function scores decreased after 6mo of ocular surface function scores were significantly lower than the preoperative (P<0. 05). While the control group after 6mo, with the passage of time, the MUC5AC content gradually reduce, ocular function score increased gradually. ?CONCLUSlON:To treat the patients with glaucoma and cataract with combined surgery, the level of MUC5AC can temporary decrease. Ocular function score can temporary increase in, but after 3mo, it can be gradually improved.

Research on polymer impurities has always been important in the quality control of cephalosporins. Research on polymers in cephalosporins that lack active amino groups on the C-7 side chain has not been reported. Therefore, our study used cefazolin sodium, which is widely used in the clinic, as an example. The polymer in cefazolin sodium and its product "cefazolin sodium pentahydrate for injection" was analyzed by column switching liquid chromatography-high resolution mass spectrometry. Two polymer impurity peaks were detected and the possible structures of these polymers were suggested. Through two-dimensional liquid chromatography, the chromatographic peaks following Sephadex gel chromatography and high-performance gel chromatography were compared to those obtained by reverse high-performance liquid chromatography (HPLC) for cefazolin sodium as reported in the Chinese Pharmacopoeia. The HPLC method proves more suitable for polymer detection than Sephadex gel chromatography and high-performance gel chromatography. The method of polymer detection for cefazolin sodium was established using the method of related substances HPLC as described in the Chinese Pharmacopoeia.

OBJECTIVEAn epidemic of severe acute respiratory syndrome (SARS) hit Beijing, China, between March and July 2003 with an attack rate of 1.9 per 10 thousand. (2,521 cases). To control the epidemic of SARS, a total of 30,173 residents were quarantined either in their residence or in quarantine sites. In order to understand the personal need of being quarantined and to estimate the risk of developing SARS during the quarantine period, a survey on the quarantined residents of Haidian District, Beijing, China was carried out.

METHODS33 precincts in Haidian District divided into five groups (7 in north, 6 in south, 7 in west, 6 in east and 7 in central of Haidian District) according to the location of the precincts were involved. The director of Center of Disease Control and Prevention of Haidian District was asked to select 1 precinct from each group according to the workload of the precinct quarantine officers. From those 5 precincts we obtained lists of all quarantined persons from the precinct quarantine officers. All quarantinees were asked to complete a self-administered questionnaire. The SARS patients were diagnosed and verified according to the diagnosis criteria released by Chinese Ministry of Health which was equivalent to the SARS 'probable case' definition of WHO. All SARS cases had been reported to the relative authorities.

RESULTSBy May 23, 2003, 5,186 persons had been quarantined in Haidian district, accounting for 0.23% of all residents. 1,028 of sampled quarantined residents completed the questionnaire. Of those who completed the questionnaire, 2.3% (95% CI: 1.5% - 3.5%) developed SARS while under quarantine. The median quarantine period was 14 days (range 1 day to 28 days). 61% of the quarantinees had a direct contact history with SARS patients, and all secondary SARS patients developed through contact to these quarantinees. The remaining 39% quarantinees who did not have a direct contact history with SARS patients had not developed SARS during the period under quarantine. 37% of the quarantees had direct contact during original patients' symptomatic period with an AR of 6.3%. Persons who looked after the illed SARS patient(s) during their symptomatic period, had an highest attack rate of 31% (95% CI: 20% - 44%). 63% (95% CI: 60% - 66%) of the total quarantined persons did not have direct contact with a SARS patient during his/her symptomatic period, with an attack rate of 0% (95% CI: 0% - 0.73%).

CONCLUSIONOnly those persons having direct contact with ill SARS patients need to be under quarantine. The overall cost for quarantine on SARS prevention could be reduced by as much as 63% if the quarantine program was limited to this group. No evidence was found that SARS patient was infective during the incubation period.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; prevention & control ; Female ; Humans ; Infant ; Male ; Middle Aged ; Quarantine ; methods ; organization & administration ; psychology ; Risk Factors ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; psychology ; Time Factors

OBJECTIVETo examine the expression of tissue factor pathway inhibitor-2 (TFPI-2) in gallbladder cancer (GBC) and to investigate the anti-cancer activities of TFPI-2 against the growth of GBC.

METHODSTFPI-2 expression in gallbladder normal tissues, gallbladder polyp (GBP) tissues and GBC tissues were examined by reverse transcriptase polymerase chain reaction (RT-PCR), Western blot and immunohistochemical staining. Adenovirus carrying human TFPI-2 gene (Ad5-TFPI-2) were constructed and its anti-cancer effects were investigated in xenograft tumors. Xenograft tumors were constructed by injection of GBC-SD and SGC-996 cells into the flank of nude mice and the volume of xenograft tumors was measured every 3 days until the sacrifice of mice. The apoptosis index of xenograft tumors was examined by TUNEL assay. The status of Bax, Bcl-2 and caspase-3 was examined by Western blot assay.

RESULTSTFPI-2 expression was profoundly lower in GBC tissues (87.0%) when compared to normal tissues (23.3%) and GBP tissues (52.2%; χ(2) = 21.104, P = 0.000). Ad-TFPI-2 significantly inhibited the growth of xenograft tumors in nude mice. Ad-TFPI-2 inhibited GBC-SD cell growth through the induction of apoptosis. The means of total apoptotic cells per field were much higher in Ad5-TFPI-2 group than those in PBS and Ad5-GFP groups. Ad5-TFPI-2 elevated the expression of Bax and cleaved caspase-3, while it decreased the expression of Bcl-2.

CONCLUSIONSTFPI-2 gene and protein was down-regulated in GBC and the down-regulation of TFPI-2 may play a role in the tumorigenesis of GBC. Adenovirus-mediated TFPI-2 can inhibit GBC growth through the induction of apoptosis.

Adenoviridae ; genetics ; Aged ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; therapy ; Genetic Therapy ; Glycoproteins ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.

METHODSGenomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.

RESULTSThe sequencing results showed HLA-DRB1 alleles of the proband as DRB1*090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1*120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.

CONCLUSIONThis allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis

BACKGROUNDSuicide gene therapy is a widely used molecular treatment for head and neck cancer. In this study, we try to use the method of homogenous recombination in bacteria to clone thymidine kinase gene (tk)-a kind of suicide gene to adenovirus backbone vectors for the construction of replication-defective adenoviruses.

METHODSpAdTrack-CMV/tk was constructed through subclone of a restriction endonuclease fragment including thymidine kinase gene from plasmid pCMV-tk to another plasmid pAdTrack-CMV, and then co-transfected with supercoiled pAdEasy-1, which was an adenoviral backbone vector except for deletions of E1 and E3, to competent E. coli BJ5183 for homogenous recombination using electroporation procedure. With the same method, pAdTrack-CMV was also co-transformed with pAdEasy-1 for homogenous recombination in BJ5183. Identified with restriction endonuclease PacI and polymerase chain reaction (PCR), plasmids pAd-GFP/tk and pAd-GFP were successfully constructed. Each of them was digested with PacI and sequently transfected into human embryo kidney 293 cells (HEK293) using Lipofectamine 2000.

RESULTSComet-like adenovirus-producing foci of Ad-GFP/tk and Ad-GFP were observed after 5 to 7 days of cell culture. After twelve days of packaging, the replication-defective adenoviruses were collected. Identified with PCR, thymidine kinase gene was successfully constructed into Ad-GFP/tk.

CONCLUSIONThe replication-defective adenoviruses containing thymidine kinase can be constructed more easily by homogenous recombination in bacteria than conventional techniques.

Adenoviridae ; genetics ; Bacteria ; genetics ; Defective Viruses ; genetics ; Genetic Therapy ; methods ; Green Fluorescent Proteins ; genetics ; Recombination, Genetic ; Thymidine Kinase ; genetics ; Virus Replication

To increase drug concentration in the head through intranasal administration, we have investigated the excised animal nasal mucosa permeability and nasal toxicity of the baicalin drug carrier systems, such as baicalin liposomes, beta-cyclodextrin inclusion compound, and phospholipid complex. A transport of baicalin drug carrier systems through nasal mucosa was simulated in diffusion chamber in vitro, and swine, caprine and rabbit nasal mucosa was used, the concentration of drug in the receptor was determined by HPLC. By taking the apparent permeability coefficients as evaluation standard, investigated the isolated animal nasal mucosa permeability of different baicalin drug systems was investigated for screening the best baicalin drug carrier system through nasal cavity administration. Toxicity of baicalin and its phospholipids complex on toad palate mucosal cilia movement and rats nasal mucosa long-term toxicity were studied in vivo. The apparent permeability coefficient of three kinds of baicalin drug carrier systems was better than that of baicalin (P < 0.05), and its lag-time was obviously shortened. At the same time, the apparent permeability coefficient of phospholipid complex was higher than those of other two drug carrier systems (P < 0.05). The results showed that the baicalin phospholipids complex nasal mucosa permeability was obviously superior to the other two drug systems. Baicalin phospholipids complex had no toxicity to ciliary movement, and had no irritation to rat nasal mucosa. The results show that baicalin phospholipid complex was the best baicalin drug carrier system, it could significantly enhance the permeability of baicalin across nasal mucosa, had no toxicity to nasal mucosa, and could be used for intranasal administration.
Administration, Intranasal ; Animals ; Bufo bufo ; Drug Carriers ; pharmacokinetics ; toxicity ; Drug Delivery Systems ; Female ; Flavonoids ; administration & dosage ; pharmacokinetics ; toxicity ; Goats ; Liposomes ; pharmacokinetics ; toxicity ; Male ; Nasal Mucosa ; drug effects ; metabolism ; Palate ; drug effects ; Permeability ; Phospholipids ; pharmacokinetics ; toxicity ; Rabbits ; Random Allocation ; Rats ; Swine ; beta-Cyclodextrins ; pharmacokinetics ; toxicity

The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B*4061, in Chinese population. DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing. The sequencing results showed HLA-B alleles of the proband as B*4601 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ089628, DQ089629, DQ089630). After HLA blast analysis, the novel allele showed a single nucleotide mismatch with B*400101 in exon 2 at position 272 C-->A, as the results, changing amino acid from Ser to Tyr at codon 67. It is concluded that this allele is a novel one and has been officially named B*4061 by the WHO Nomenclature Committee.
Alleles ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; HLA-B Antigens ; genetics ; immunology ; Histocompatibility Testing ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo do some comparative study on anti-inflammatory and antipyretic effects between the Dao-di herb and non Dao-di herb of Huangqin (the roots of Scutellaria baicalensis) and provide thinking and evidence for study on geoherbalism and clinical usage of Huangqin.

METHODThe anti-inflammatory action was assessed by auricular swelling induced by dimethylbenzene in mice and the antipyretic action was monitored by dried yeast-induced mice fever.

RESULTAll samples of both Dao-di and non Dao-di herbs of Huangqin showed antipyretic effect. The Dao-di Huangqin samples showed antipyretic effect between 61% to 53% , whereas the non Dao-di Huangqin samples between 53% to 43%. Six Dao-di Huangqin samples showed better antipyretic effect than four non Dao-di Huangqin samples. All samples of both Dao-di and non Dao-di Huangqin showed anti-inflammatory effect. Dao-di Huangqin showed anti-inflammatory effect between 73% to 54%, whereas non dao-di Huangqin between 53% to 34%. Six Dao-di Huangqin showed better anti-inflammatory effect than four non Dao-di Huangqin. In totality, results from analysis of geoherbalism showed that geoherbal production areas of Huangqin had better effect than that of the non geoherbal production areas in anti-inflammatory and antipyretic effects.

CONCLUSIONBoth the Dao-di and non Dao-di Huangqin have effects of anti-inflammatory and antipyretic to a certain extent, but the efficacy of the Dao-di Huangqin surpass the non Dao-di Huangqin.

Animals ; Anti-Inflammatory Agents ; administration & dosage ; Antipyretics ; administration & dosage ; China ; Drug Contamination ; Drugs, Chinese Herbal ; administration & dosage ; Fever ; drug therapy ; Humans ; Inflammation ; drug therapy ; Mice ; Scutellaria baicalensis ; chemistry

OBJECTIVE Paeoniflorin (PF) and albiflorin (AF) are the major active components of total peony glucosides(TPG)from Paeonia lactiflora Pal,which have many biological activities such as anti-inflammatory, antioxidation and anti-hypertension effects. The drug-drug pharmacokinetic interaction among PF,AF and TPG,the pharmacokinetic comparisons of AF between hypoxia and normoxia,the transport of AF cross the blood-brain barrier cell model and the transport of AF/PF/TPG cross Caco-2 cell model were investigated.METHODS A highly sensitive and rapid UPLC-MS method with multiple-reaction monitoring(MRM)scanning via electrospray ionization(ESI)source operating both in the positive and negative ionization mode was successfully developed and validated for simultaneous quantitation of PF and AF in rat plasma after an oral administration of PF,AF and TPG. RESULTS The validated and developed UPLC-MS/MS method was successfully applied to simultaneously determine the AF and PF concentration in rat plasma and investigate pharmacokinetic interactions after a single intragastrical ad-ministration of PF,AF,co-administration of PF with AF and TPG,respectively.The elimination of both PF co-administered with AF and PF in TPG were slower than those for PF alone and the distribution in the tissues was wider.The combination of PF with AF or TPG could significantly increase the values of the AUC, MRT and t1/2of the drug PF, and reduce the values of CL of PF. From a comparison of the main pharmacokinetic parameters among AF alone, AF combined with PF and AF in TPG, the values of the MRT and t1/2of AF in TPG were greater than that of AF alone,and there were statistically signifi-cant differences in these parameters(P<0.05,P<0.01).It was also noticed that AUC and Cmaxof PF in hypoxia rats were significantly decreased compared with that of normaxia rats, suggesting that there was a decreased exposure of PF in rats under hypoxia. The multiple active components in TPG may lead to DDIs between some P-gp substrates. CONCLUSION The clinical performance of total peony glucosides would be better than that of single constitute. The outcomes of the study are expected to serve as a basis for development of clinical guidelines on total peony glucosides usage.