Objective: To develop an HPLC-MS method for the determination of phillyrin, phillygenin, forsythiaside A, forsythiaside F, rutin, quercetin, isoquercitrin, chlorogenic acid, and (+)-epipinoresinol-4'-O-glucoside in Forsythia Folium. Methods: The samples were separated on a Dikma Diamonsil C18 column (150 mm × 4.6 mm, 5 μm), eluted with methanol and water containing 0.02% formic acid in a gradient elution at a flow rate of 0.8 mL/min, and the injection volume was 10 μL. The column temperature was set at 30°C. The multiple-reaction monitoring scanning was employed for the quantification with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at -4500 V and the turbo spray temperature was maintained at 650°C. Results: The regression equations showing linear relationships between peak areas and contents of each compound were obtained. The average recoveries of the compounds ranged from 97.20% to 101.2% and the precision in terms of RSD was in the range of 0.54%-1.24%. Conclusion: The method is sensitive and suitable for the determination of the nine components in Forsythia Folium. As a result, the suitable harvesting time for Forsythia Folium is May.

Background Trachoma is a common disease in ophthalmology,but few reports about the pathogenetic genotype are reported.Objective This study was to investigate the genotypes of Chlamydia trachomatis.Methods Conjunctival specimens were collected in 16 patients with trachoma in Beijing Tongren Eye Center from January,2003 and August,2006.Variable sequence 4 (VS4) of ompl fragment was amplified by nested PCR(n-PCR) using specific primer of Chlamydia trachomatis,and then the PCR products were sequenced.The DNA sequences were analyzed by software Clustal X and MEGA2,and the genetic characteristics were compared with the known sequences of GenBank.Results The PCR product fragment of MOMP gene of trachoma was 277 bp.Three types of Chlamydia trachomatis strains were idcntified in the 16 specimens,including B-genotype in 9 strains (56.3％),C-genotype in 4 strains(25.0％) and D-genotype in 3 strains(18.7％).High homology was seen in the gene sequences of Chlamydia trachomatis strains between the B-genotype or C-genotype strains and the same genotypes of GenBank or those from some districts of China.However,some differences were exhibited among the Dgenotype strains.Conclusions Identification of genotype of Chlamydia trachomatis has differential effect on trachoma and inclusion conjunctivitis.

Objective To investigate the clinical manifestation of trachoma,and to select the appropriate laboratory test for clini- cal diagnosis.Design Retrospective case series.Participants Retrospective analysis of medical records from 61 patients with trachoma from Jan 2003 to Aug 2006 in Bejing Tongren Eye Center.Methods Grades of trachoma diagnosis were according to the criteria de- signed by Chinese Ophthalmological Society (1979).The general state of health,case history,and the laboratory investigations of pa- tients were recorded.Laboratory tests included the conjunctiva scraping for inclusions,C.trachomatis immune antigen test and PCR test.Main Outcome Measures Manifestation of corneal and conjunctiva,the results of laboratory tests of C.trachomatis.Results Out of sixty-one patients including 28 males and 33 females,the average old was (29.05?19.99) years.88.5% cases were inⅠstage of tra- choma,8.2% were inⅡstage,and 3.3% were inⅢstage.The C.trachomatis inclusions were found in 7 (11.5%) scraping smears.42 (68.9%) cases were positive in C.trachomatis antigen test.46 (75.4%) cases were positive in PCR tests.The positive rates of antigen and PCR test were significantly higher than that of scraping (P=0.00).Conclusions A majority of clinical patients were inⅠstage of trachoma.The degree of their distress was minimal.It is necessary to apply C.trachomatis,antigen test or PCR test to improve the clin- ical diagnosis.

Objective To study the characteristics of Genetic typing and the antibiotic susceptibility testing of strains from Pseudomonas aeruginosa keratitis patients.Design Experimental study,Participants 23 eyes of 23 patients of Pseudomonas aeruginosa keratitis.Methods The genomic was extracted and amplified with PCR.The PCR products were purified and sequenced.The results were registered in MIST web Antibiotic susceptibility testing were performed in theses strains,Main Outcome Measures Sequence types and antibiotic susceptibility.Results The isolates were resolved into 20 STs.Two lineages were identified.MIC test showed that strains were more susceptible to aminoglycosides,The activity of quinolones and cephalosporin were higher than that of aminoglycosides.Conclusion MIST can determine homology of the strains from Pseudomonas aeruginosa keratitis by clustering results. There is no finding about relationship between Genetic typing and drug resistance.

OBJECTIVETo observe the effect of Qingfei Peiyuan Micro-pill (QPM) on HIV/AIDS patients with pulmonary infection of phlegm heat obstructing lung syndrome (PHOLS).

METHODSTotally 141 HIV/AIDS patients with pulmonary infection of PHOLS were randomly assigned to the treatment group (94 cases) and the control group (47cases). On the basis of Western medicine, patients in the treatment group took QPM. The therapeutic course for all was 28 days. The improvement of symptoms and signs was observed. The body temperature (BT), chest X ray, and white blood cells (WBCs) were detected.

RESULTSThe Chinese medical syndrome score was lower in the treatment group than in the control group at the 7th, 21st, and 28th day of treatment, showing statistical difference (P < 0.05). The efficacy was better in the treatment group than in the control group at the 7th, 21st, and 28th day of treatment, showing statistical difference (P < 0.05). The BT was lower in the treatment group than in the control group on the 7th day. There was no statistical difference in the patient number with normal WBCs on the 7th day (P > 0.05). But there was statistical difference in the patient number with normal WBCs on the 14th, 21st, and 28th day of treatment (P < 0.05). There was no statistical difference in the patient number with normal chest X ray on the 7th and 28th day of treatment (P > 0.05). But there was statistical difference in the patient number with normal chest X ray on the 14th and 21 st day of treatment (P < 0.05).

CONCLUSIONQPM had certain complementary effect on HIV/AIDS patients with pulmonary infection of PHOLS.

Acquired Immunodeficiency Syndrome ; complications ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Respiratory Tract Infections ; complications ; drug therapy ; Treatment Outcome

In the current study, a total of nineteen triterpenoids (1-19) from 60% EtOH extracts of Stauntonia obovatifoliola Hayata subsp. intermedia stems were separated and purified by solvent extraction and chromatographic methods including silica gel, ODS as well as preparative HPLC. According to the results of chemical reactions and spectral data, compounds were identified as: lupeol (1), betulinonic acid (2), betulinic acid (3), 3-epi-betulinic acid (4), quinatic acid (5), 24-O-acetyl quinatic acid (6), 3-O-α- L-arabinopyranosyl-30-nor-hederagenin-28-O-α-L-rhamnopyranosyl-(1 --> 4) -β-D-glucopyranosyl-(1 --> 6) -β-D-glucopyranosyl ester (7), Stauntoside A (8), kalopanax saponin A (9), kalopanax saponin J (10), Kizuta saponin K10 (11), 3-O-α-L-rhamnopyranosyl (1--> 2) -α-L-arabinopyranosyl-hederagenin-28-O-β-D-xylopyranosyl-(1 --> 6) -β-D-glucopyranosyl ester (12), kalopanax saponin B (13), 3-O-α-L-rhamnopyranosyl-(1 --> 2) -α-L-arabinopyranosyl-hederagenin-28-O-β-D-glucopyranosyl-(1 --> 6) -β-D-glucopyranosyl ester (14), sieboldianoside A (15), septemoside A (16), kalopanax saponin K (17), septemloside I (18), and 3-O-α-L-arabinopyranosyl (1 --> 2)-β-D-glucuronopyranosyl- hederagenin (19). Among them, compounds 4, 6, 10, 12, 14, and 16-19 were isolated from the Stauntonia genus for the first time, and compound 6 was a new natural product.
Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Magnoliopsida ; chemistry ; Molecular Structure ; Plant Stems ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry

Objective To characterize the clinical manifestations, laboratory findings of patients with occult sys?temic malignant neoplasms, whose initial manifestation presented as multiple acute cerebral infarcts including coagula?tion function,radiological imaging and microembolic signals (MES) detection by transcranial Doppler sonography (TCD) and to explore the possible underlying mechanisms. Methods All clinical records, laboratory hematological tests includ?ing hypercoagulable states measured by D-dimer levels, brain MRI including DWI, and TCD monitoring MES, the treat?ment and prognosis were retrospectively reviewed in 12 patients with multiple acute cerebral infarcts as the first manifes?tation of occult systemic malignancy. Results The clinical manifestations presented as localized neurological dysfunction, e.g. hemiparesis, aphasia, hemiparesthesia, dysarthria, vertigo and seizures, etc. DWI revealed multiple disseminated acute cerebral infarcts in multiple arterial territories such as the bilateral anterior or anterior plus posterior cerebral circu?lation in all patients. Eleven of 12 patients tested had elevated D-dimer. TCD detected MES in 5 of 7 patients. There were 12 patients diagnosed with occult systemic malignancy including 5 lung cancer, 3 pancreatic cancer, 1 gastric can?cer, 1 colon cancer, 1 endometrial adenocarcinoma and 1 metastatic poorly differentiated mucinous adenocarcinoma with unknown primary. Ten patients already had remote metastasis at diagnosis. The prognosis was usually poor and there were 7 cases with ischemic stroke recurrence, 4 cases with acute myocardial infarction, 3 cases died during hospitaliza?tion. Conclusions When patients present with multiple disseminated acute cerebral infarcts involving multiple arterial territories as initial manifestation, the underlying occult systemic malignancy should be considered. Hypercoagulopathy and MES might provide the clues to the diagnosis.

BackgroundStudies confirmed that ultraviolet A (UVA)- riboflavin photodynamic therapy can control keratoconus progresses by altering the physicochemical property of cornea.The collagen components of amniotic membrane transplantation is similar to that of cornea and amniotic membrane transplantation has been widely used to ocular surface reconstruction.However,the study on UVA riboflavin-induced-collagen crosslinking for amniotic tissue is less now.ObjectiveThis study was to investigate the role of UVA-riboflavin on frozen-preserved human amniotic membrane.Methods Human amnions were obtained in informed consent and prepared into 2 mm×15 mm pieces and were then divided into 4 groups using lottery method and 6 pieces for each group.The first 3 groups were treated with the photosensitizer riboflavin and UVA-irradiation ( wavelength:370 nm ; irradiation energy:1,2 or 3 mW/cm2,distance:10 mm) for 30 minutes,and the untreated fourth group was as control group.Biomechanical stress-strain test was performed using a microcomputer-controlled biomaterial tester and the stress(mN) was recorded when the strains were set to 5％,10％ and 15％.The 7 mm diameter of human amniotic membrane pieces were trephined and divided into 4 groups(5 pieces for each group) with the treated method as mentioned above,and then the buttons were exposed to 0.1％ collagenase Ⅰ solution.The transparency was scored and the complete dissolving time was record.In histological evaluation,three groups (3 pieces for each group) of human amniotic membranes were treated using UVAriboflavin(3 mW/cm2),0.1％riboflavin,normal saline for 30 minutes respectively and examined under the transmission electron microscopy.This study was performed under the permission of the Ethic Commission of Beijing Tongren Hospital.ResultsWhenthestrainwas 5％,10％,15％,thestressof controlgroupand1,2,3 mW/cm2UVA group were statistically signifcantly different ( F =3.411,P =0.037; F =9.927,P =0.001;F=11.118,P=0.000).The tensile strength of human amniotic membrane cross-linked with UVA-riboflavin was statistically significantly increased in comparison to the control group (P＜0.05 ),and the tensile strength of human amniotic membrane became stronger as UVA power increased.The complete dissolve time was (8.6± 1.8 ) hours for the control group,(39.6± 2.3 ) hours for 1 mW/cm2 UVA group,(71.4±0.9 ) hours for 2 mW/cm2 UVA group,(78.8± 1.8 ) hours for 3 mW/cm2 UVA group,showing the enhanced anti-enzyme ability of human amniotic membrane after cross-linking(P＜0.01 ).The collagen density in the UVA-riboflavin treated group was increased,the connection among the collagen fibers as well as between the stroma and the epithelium became tighter than those of control group.ConclusionsCollagen cross-linking with UVA-riboflavin make the biomechanical strength and enzymatic resistance of human amniotic membrane enhance and ultrastructure change of human amniotic membrane.

Objective To analyze clinical diagnosis and management of 5 patients with actinomycete keratitis.Design Retro- spective case series.Participants 5 patients (5 eyes) with actinomycete keratitis.Methods The clinical features and microbiologic da- ta of 5 culture-proven cases of actinomycete keratitis recorded between October 2004 to March 2006 were analyzed.Main Outcome Measures clinical characteristics,isolations identification,drug susceptibility test and treatments.Results All patients were males and farmers.Of the 5 cases presented in this study,4 cases were followed by minor trauma as a predominant risk factor,and were pre- sented by a chronic progressive corneal ulcer with a wreath pattern of infiltrate.The diagnosis of all cases was based on laboratory in- vestigations,by which 4 cases of nocardia and one case of streptomyce were identified.A variable drug sensitivities were presented in nocardia isolates,which including TMP-SMZ,amicasin,gentamicin and fluorine-quinolones.Conclusions Nocardia keratitis is mainly followed by a minor trauma.It is identified predominantly by laboratory investigations.Tropical and systemically sensitive biotic are the initial choice,while debridement and amnionic transplantation could be an effective alternative.

Wuzhuyu Tang is a classical formula for treating migraine, but its' pharmacological ingredients is unclear yet. Present study employed the everted intestinal sac model to collect the absorption samples of 10 kinds of Wuzhuyu decoction, and then analyzed the contents of 9 ingredients in Wuzhuyu Tang and absorption samples quantitatively or semi-quantitatively by HPLC-DAD method. Reserpine was used to establish the mice model of migraine, and then the contents and activities of 5-hydroxytryptamine, noradrenaline, dopamine, nitric oxide and nitricoxide synthase in brain tissues and serums were determined respectively after oral administration of Wuzhuyu Tang. Using the partial least squares regression method to correlate the total absorption quantity of 9 ingredients and pharmacodynamics. The result shows that limocitrin-3-O-beta-D-glucoside, ginsenoside Rg1 and Rb1, rutaevine, limonin, evodiamine and rutaecarpine are the main ingredients influenced the effects in absorption samples in everted intestinal sacs, especially ginsenoside Rg1, rutaevine, evodiamine and rutaecarpine among them have obvious improving effects to most pharmacodynamics index, might be the pharmacological ingredients influenced the therapeutical effects of Wuzhuyu Tang treating migraine.
Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; Male ; Mice ; Mice, Inbred ICR ; Migraine Disorders ; drug therapy ; Rats ; Rats, Wistar