Objective: A novel method for the analysis of alkaloids in Nelumbo nucifera leaves was established by SPE-RRLC-Q-TOF mass spectrometry. Methods: The crude sample was extracted by 1% HCl with ultrasound-assisted extraction, then purified by SPE column, and eluted with ammonia-methanol (5:95). After concentration, the residue was dissolved by methanol solution. The real sample was analyzed by RRLC-Q-TOF. A Welch Materials C18 column was applied in the RRLC separation using acetonitrile and water as mobile phase. The elutes were detected by Q-TOF to obtain the MS spectra with extract molecular weights under positive ion mode. Results: Nine alkaloids were identified. Conclusion: This method can be used to rapidly determine the alkaloids of N. nucifera leaves.

Objective: A novel method for the separation of alkaloids from Nelumbinis Folium by pH-zone-refining counter-current chromatography (pH-ZRCCC). Methods: The two-phase solvent system composed of MtBE-methanol-water (4:1:5), 10 mmol/L triethylamine (TEA) in organic stationary phase and 5 mmol/L HCl in aqueous mobile phase. The upper phase was used as the stationary phase, while the lower phase was used as the mobile phase. In one-step elution under the conditions of a flow rate of 1.5 mL/min, 800 r/min and the effluent was detected at 254 nm. Results: From 2.18 g of the crude extract, 110 mg N-nornuciferin, 420 mg O-nornuciferin, 310 mg nuciferine, and 190 mg roemerine were obtained and each with a purity of over 98% as determined by HPLC. The structures of the isolated compounds were identified by HPLC, MS, and 1H-NMR. Conclusion: The present pH-ZRCCC method may be applied to the purification of other various alkaloids from natural products.

Objective To explore the possible association of single nucleotide polymorphism (SNP) rs3738435 of muscarinic acetylcholine receptor subtype M3 gene (cholinergic receptor, muscarinic 3, CHRM3) with risk of type 2 diabetes mellitus (DM) and metabolic disturbance. Methods The genotypes of T-149C variant of CHRM3 gene were determined by PCR-RFLP in 573 Chinese individuals in Shanghai, including 220 newly-diagnosed type 2 DM patients without taking any drug and 353 subjects with normal glucose tolerance (NGT). In the subjects, height and weight were measured for body mass index(BMI), waist, hip and femoral circumstances for waist-to-hip ratio (WHR) and waist-to-femur ratio (WFR), and serum lipid level including total cholesterol, triglyceride, high-density and low-density lipoprotein cholesterol, blood pressure, plasma glucose levels both at 0 and 120 minute during oral 75 g glucose tolerance test (OGTT) were also determined. Results (1) There was no statistical difference in the gene frequency between groups of type 2 DM and NGT. (2) In the group of type 2 DM, significant differences were observed between TT genotype carriers and TC+CC genotypes carriers for BMI, with an obvious increase in TY genotype carriers [(26.99±3.59vs25.34±3.48)kg/m2, P=0.001]. (3) In the subgroup of type 2 DM with BMI≥25 kg/m2, total cholesterol was higher in TT genotypes than in TC+CC genotypes[(5.75±1.26vs5.27±1.14)mmol/L, P=0.030], so was the low-density lipoprotein cholesterol. Conclusion The genetic variation T-149C in the CHRM3 gene seems to attribute to weight regulation and lipid metabolism of patients with type 2 diabetes mellitus in Chinese population.

AIM: To report a two- year's results of iontophoresis-assisted transepithelial corneal cross-linking (I-CXL) for progressive keratoconus.
METHODS: Thirty - four eyes in 24 patients with progressive keratoconus ( mean age 21. 0 ± 5. 6 years;range: 14-32 years) were treated. After 1g/ L riboflavin-distilled water solution was administered by iontophoresis-assited (current 1mA) transepithelial method for 5min in total, standard surface UVA irradiation ( 370nm, 3mW/cm2 ) was performed at a 1 - cm distance for 30min. The best corrected visual acuity ( BCVA ) measured as LogMAR number, corneal refractive astigmatism, K1, K2, Kmean, Kmax, intraocular pressure, endothelial cell density, the thickness at corneal apex and the thinnest point were measured preoperatively and 2a postoperatively.
RESULTS:At 2a after the procedure, BCVA (LogMAR) improved from 0. 32 ± 0. 25 to 0. 25 ± 0. 19 ( t = 2. 849, P =0. 015). K1 decreased from 47. 12±4. 33 to 46. 06±4. 77 (t =2. 652, P= 0. 015). K2 decreased from 51. 36±5. 59 to 50. 40±6. 16 (t= 2. 121, P= 0. 047). Kmean decreased from 49. 12±4. 76 to 48. 10±5. 25(t = 2. 663, P = 0. 015). Kmax decreased from 57. 57±8. 30 to 55. 91±8. 14 (t = 2. 398, P = 0. 026). The corneal apex thickness decreased from 476. 90±38. 71μ m to 454. 43 ± 40. 86μ m ( t = 2. 853, P = 0. 010 ). The thinnest thickness decreased from 464. 38 ± 39. 92μ m to 433. 86 ±50. 78μ m ( t = 3. 485, P = 0. 002 ). Corneal refractive astigmatism, intraocular pressure and endothelial cell density did not show significant changes.
CONCLUSION: I - CXL for progressive keratoconus is safe and effective which can prevent deterioration of progressive keratoconus within 2a, but further long-term studies are necessary still.

Objective To detect mutations of the ADAR1 gene in three Chinese families with dyschromatosis symmetrica hereditaria (DSH).Methods DNA was extracted from the blood samples of seven patients with DSH and their 33 relatives in three families with DSH as well as from 50 unrelated healthy controls.PCR and direct sequencing were performed to detect mutations in the ADAR1 gene.Results All the patients carried mutations in the ADAR1 gene.Three mutations were identified,including one frameshift mutation c.2433-2434delAG in family 2 and two missense mutations,i.e.,c.1760A ＞ G (p.Y587C) in family 1 and c.3620G ＞ T (p.G1207V) in family 3.No mutations were found in the ADAR1 gene in unaffected individuals in these families or the healthy controls.Conclusion Two novel missense mutations are found in the ADAR1 gene of two Chinese families,which may represent a molecular mechanism underlying the development of DSH.

Objective Application of the second metatarpophalangeal joint by traction prolong trans- plant repair the defects in the metacarpophalangeal joint,reconstruct the function of it.Methods By means of the apparatus to prolong finger in advance,then transplant the second metatarpophalangeal joint to recon- struct metacarpophalangeal joint for seven cases of obsolete defects in the metacarpophalangeal joint.Results The average of finger prolong was 2.6 cm,consultation from 1 to 4 years.average 2.5 years,thai the trans- plant joints have all survived and osteal concrescence.Through the criterion Chinese Medical Association,good rate was 85.7%. Conclusion It' s a good method to repair obsolete defects in the metacarpophalangeal joint by transplant traction prolong of the second metatarpophalangeal joint.

OBJECTIVETo observe the clinical efficacy of Tiaozhi Jiangtang Tablet (TZJT) on patients suffering from diabetes mellitus with blood stasis syndrome.

METHODSSixty patients were randomly assigned into 2 groups, the TZJT group (n = 30) treated with TZJT tablet and asprin, the control group (n = 30) treated with asprin alone.

RESULTSThe improvement of symptoms was more significant in the TZJT group than that in the control group (P < 0.05). Levels of serum endothelin (ET), nitric oxide (NO) content and blood viscosity were decreased in both groups after treatment, and the effect of TZJT plus asprin was superior to that of asprin alone.

CONCLUSIONTZJT combined with asprin is effective in improving the serum content of ET and NO and reducing blood viscosity.

Adult ; Aged ; Aged, 80 and over ; Coronary Artery Disease ; drug therapy ; etiology ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

Objective:To investigate the heterogeneity of epitopes recognized by anti-GBM autoantibodies in sera from a large cohort of Chinese patients with anti-GBM disease and its clinical significance.Methods: The present study included 108 patients with anti-GBM disease who were diagnosed in our hospital, between Jan 1991 and May 2009, with complete clinical and renal pathological data. Sera or plasma exchange of the patients were used to incubate with cryostat section of normal human renal tissue for indirect immunofluorescence (IIF) assay. The cryostat sections of normal renal tissue were pre-treated by 6 mol/L urea to unmask cryptic epitopes, and untreated cryostat sections were used to detect natural exposed epitopes. The sera were diluted from 1:2 to 1:512 to determine titers of anti-GBM autoantibodies Patients with anti-GBM autoantibodies against cryptic or exposed epitopes were further stratified;their clinical and pathological associations were analyzed. Results: Sera from all the 108 patients could recognize cryptic epitopes on normal renal tissue ( urea treated section). IIF showed IgG linear staining along GBM. However, sera from 56/108 patients (group A) could also recognize exposed epitopes on normal renal tissue (untreated section) ; sera from the rest 52/108 patients (group B) could not recognize exposed epitopes. In urea treated condition, the average titer of anti-GBM autoantibodies from sera of patients in group A was significantly higher than that in group B (P＜0.01) , ANCA-positive patients in group A were significant less than that in group B (P＜0.01) . There was no significant difference between the two groups in regard to other clinical data (including serum creatinine) and renal histopathologic data. Conclusion: Anti-GBM autoantibodies from some patients with anti-GBM disease could recognize natural exposed epitopes, however, their anti-GBM titer for cryptic epitopes was higher than that of those recognizing cryptic epitopes only and the prevalence of serum ANCA was significantly less.

Objective To explore the polymorphism of HLA class I alleles of HIV-infected former plasma donors and to investigate its impact on HIV-1 viral load in central China.Methods 106 subjects chronically infected with HIV-1 were recruited and HLA class I alleles were genotyped with PCR-SSP assay.HLA class I genotypes and haplotypes were determined and their association with plasma viral loads were analyzed.Gag-specific CTL responses were detected by an IFN-λ EUSPOT assay by using overlapping peptides,and their association with plasma viral loads were also analyzed.Results Subjects homozygous at HLA class I locus had higher plasma viral loads(P=0.0098);HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads(P=0.0004,0.0103,0.0058,0.0371 and 0.0006);an inverse correlation between p2p7p1p6-specific CTL responses and viral loads in subjects with HLA-A*30/B*13/Cw*06 haplotype as well as an inverse correlation between p17-specific CTL responses and viral loads in subjects with HLA-Cw*14 allele were observed.Conclusion HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads and Gag-specific CTL responses restricted by these HLA alleles may contribute to the protection.

Objective To learn the present status of defluoridation water improvement project in Shanxi province in order to provide scientific basis for speeding up the prevention and control of endemic fluorosis.Methods According to "The National Technical Scheme for Endemic Disease Control" from 2005 to 2009, the investigation points were selected in the counties that implemented the measures of water improvement and defluoridation,the status of drinking water defluoridation Project was investigated, and the water fluoride levels were determined by fluoride selective ion electrode. Results The primary status was surveyed in 1658 water improvement and defluoridation projects in 51 counties. The resource of drinking water for water improvement and defluoridation projects was mostly ground water[accounting for 93.12% (1544/1658)]. Among 1658 water improvement and defluoridation projects 1405 projects worked well(accounting for 84.74%) and 190 projects intermittently worked (accounting for 11.46%). Sixty three projects abandoned (accounting for 3.80%), in Datong basin the abandoned projects accounted for 36.36% (12/33). Water fluoride content of 1595 water improvement and defluoridation projects had been determined, among them water fluoride content of 874 projects were above 1.0 mg/L (accounting for 54.80%). The situations of exceeded national standard in the five basins was different(H = 33.22,P ＜ 0.01). The rate of over national standard of fluoride levels in drinking water was 88.37%(38/43) in Datong basin. Therefore, in Datong basin water improvement should be strengthened. Conclusions In Shanxi province the water improvement and defluoridation projects are basically running normally. However, the qualified rate is lower for the water improvement and defluoridation projects. The water improvement status varies dramatically among areas.The situation is still grim in Shanxi province. Water improvement and defluoridation needs to be strengthened to improve the effect of prevention and control of the disease.