Objective To establish a quantitative method to determine the content of platycodin D,D3 and E in Radix platycodi.Methods High performance liquid chromatagraphy-evaprorative light scattering dector(HPLC-ELSD) method was adopted.Platycodin D,D3 and E were separated by C18(5 ?m,4.6 mm?150 mm) with acetonitrile-water as mobile phase in a gradient program;flow rate was 1.0 mL/min,the temperature of drift tube was set at 113 ℃,and the gas flow(N2) was set at 3.0 L/min.Results The linear ranges of platycodin D,D3 and E were 13.78-275.6 ?g/mL(r=0.999 5),8.40-168.0 ?g/mL(r=0.999 7) and 12.02-240.4 ?g/mL(r=0.999 6),respectively.The average recoveries(n=5) were 98.3%,99.4% and 101.3%,respectively.Conclusion The method is convenient,accurate and reliable,and can be used for the determination of platycodin D,D3 and E in Radix platycodoi.

Background Many researches confirmed that basic fibroblast growth factor (bFGF)plays an important role on the proliferation and differentiation of retinal progenitor cells,but its low intraocular permeability limits its clinical application.To explore an effective approach to enhance the intraocular permeability of bFGF has an important significance for the treatment of retinopathy. Objective This study was to investigate the effect of azone on bFGF intraocular permeability after its topical administration. Methods Eighteen New Zealand white rabbits were randomly divided into four groups on random number table method.Distilled water( blank control group),5％ bFGF eyedrops(5％ bFGF group ),0.4％ azone+5％ bFGF eyedrops (0.4％ azone + 5％ bFGF group ) and 0.4％ azone+ 10％ bFGF eyedrops (0.4％ azone + 10％ bFGF group)were topically administered in different groups at 5- minute interval for 3 times.Aqueous and vitreous fluid were extracted 30 minutes after administration of eyedrops,and those in the 0.4％ azone + 5％ bFGF group were obtained 30,60 and 120 minutes after administration.bFGF concentration in the aqueous and vitreous fluid was quantified with ELISA. Results The bFGF levels(A value)in aqueous and vitreous fluid were 0.1007±0.0100 and 0.1340±0.0100 after topical administration of the 5％ bFGF eyedrops,those in blank control group were 0.1363 ±0.0100 and 0.1130±0.0100,respectively,and those in the 0.4％ azone+5％ bFGF group and 0.4％ azone+10％ bFGF group were significantly higher than the 5％ bFGF group ( both P=0.000).However,no significant difference was found in bFGF levels between 0.4％ azone+5％ bFGF group and 0.4％ azone + 10％ bFGF groups in both aqueous and vitreous fluid ( P =0.985,0.098 ).A value of bFGF in aqueous was gradually increased with prolong of time in the 0.4％ azone+5％ bFGF group,with the values 0.9413±0.0300 at 30 minutes,0.3865±0.0300 at 60 minutes,and 0.2550±0.0300 at 120 minutes,showing a positive linear correlation between bFGF level and time( R2 =0.736,P =0.003 ),but no significant correlation was seen in vitreous sample(R2=0.196,P=0.233). Conclusions Azone can improve the intraocular penetration of bFGF eyedrops.Increasing the concentration of bFGF in eyedrop from 5％ to 10％ dose not change its intraocular distribution.The highest content of the bFGF in aqueous is at 30 minutes following the administration of 0.4％ azone+5％ bFGF eyedrops.

Objective To investigate the effect of the nuclear factor (NF)-κBp65 antisense oligonucleotide (ASODN) on NF-κB activity and expression of interleukin(IL)-6 in hepatic stellate cells (HSC). Methods The HSC were separated from rats and cultured. The toxicity of NF-κBp65 ASODN on HSC were detected by Trypan blue exclusion staining and the NF-κB activity was determined by EMSA. The expressions of IL-6 mRNA and protein were meaured by RT-PCR and ELISA, respectively. Results In vitro, no toxicity of ASODN on HSC was observed at the concentrations of 0. 001 to 1.0 μmol/L. NF-κB activity was increased after stimulating HSC with tumor necrosis factor (TNF)α, whereas it was weakened in a dose dependent manner when HSC were cultured with ASODN (concentration from 0. 001 to 1.0 μmol/L). At the same time, the expressions of IL-6 mRNA and protein induced by TNFα were decreased after transfected with ASODN at concentrations of 0.001- 1. 0 μmol/L in a dose dependent manner. Conclusion ASODN may specifically inhibit either the activiy of NF-κB or expression of IL-6, which provides the theoretical basis that ASODN may use to treat fibrosis of the liver.

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. We have previously identified the benzoyl sulfathiazole derivative II as a non-competitive PTP1B inhibitor with in vivo insulin sensitizing effects. Preliminary SAR study on this compound series has been carried out herein, and thirteen new compounds have been designed and synthesized. Among them, compound 10 exhibited potent inhibition against human recombinant PTP1B with the IC50 value of 3.97 micromol x L(-1), and is comparable to that of compound II.
Humans ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Structure-Activity Relationship ; Sulfathiazoles ; chemistry ; pharmacology

Objective To explore the significance of surgical resection for the pineal region tumor,the extent of tumor resection,the surgical approaches and treatment measures of hydrocephalus.Methods Twentythree patients diagnosed of pineal region cancer were recruited for this study.Thirteen patients received ventriculo-peritoneal shunt(V-P),during which 7 cases received intraoperative end-plate colostomy.Two of the 13 cases received a second V-P procedure.Another 13 cases received operation by corpus callosum- septum-Dome Room-the third ventricle approach to remove the tumor; Eight cases underwent the tumor resection by suboccipital supratentorial approach (Poppen approach )and 2 cases underwent the tumor resection by the infratentorial cerebellar approach( Krause approach).Results Total tumor resection was performed in 11 cases,subtotal or major total resection in 8 cases and partial resection in 4 cases.Nine cases underwent postoperative radiotherapy alone,8 receiving radiotherapy plus chemotherapy,and 6 cases receiving neither radiotherapy nor chemotherapy.Two cases died during treatment.Conclusion Though it is not practicable to completely remove the germ cell tumor in patients with relatively large tumors,cerebrospinal fluid circulation pathways should be reestablished,including ventriculo-peritoneal shunt,colostomy from the end of the third ventricle to the end plate.If tumors are not too large,they would be easy to be removed.The effects on the surrounding brain tissue would not be much significant.If the connection of Ⅲ,V ventricles were normal,the patient may not need shunt or colostomy.Radiotherapy and chemotherapy can be given to this kind of patients after the surgery.To improve the efficacy,other types of tumors,expecially the benign tumors,should be totally removed.

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. We have previously identified the benzoyl sulfathiazole derivative II as a non-competitive PTP1B inhibitor with in vivo insulin sensitizing effects. Preliminary SAR study on this compound series has been carried out herein, and thirteen new compounds have been designed and synthesized. Among them, compound 10 exhibited potent inhibition against human recombinant PTP1B with the IC50 value of 3.97 micromol x L(-1), and is comparable to that of compound II.

Objective To investigate the clinical effect of IFNα combined with mannan peptide in treatment of patients with HBeAg-positive chronic hepatitis B ( CHB ). Methods Eighty HBeAg-positive CHB patients with HBV DNA quantity ranging from 10 to 10 eopies/mL were enrolled and randomized into the treatment group and the control group ( n = 40 for each ). Patients in treatment group were given daily subcutaneous injection of IFNα-2b 5,000,000 U for 52 weeks, and received mannan peptide 10 mg per intravenous injection or 2. 5 mg per intramuscular injection for a total of 2 to 3 treatment courses (12 weeks for each). The control group received only IFNα-2b treatment. Liver function, serum markers of hepatitis B, HBV DNA quantity and blood tests were performed before the treatment and at 2, 4, 8, 16, 26 and 52-week during the treatment; and the adverse effects were recorded. Results The rates for ALT normalization, negative HBsAg, negative HBeAg, HBeAg seroconversion and negative HBV DNA were 91. 8% , 17. 5% , 52. 5% , 27. 5 % and 47. 5% at 52nd week in the treatment group, while those in the control group were 80. 0% , 12. 5% , 30. 0% , 10. 0 % and 25. 0% , respectively. There were significant differences in HBeAg-negative, HBeAg-seroeonversion and HBV DNA-negative rates between two groups (χ2 = 4. 178, 4.021 and 4.381, P < 0. 05 ) , and these indexes in the treatment group were increased to 57. 5% , 30. 0% and 50. 0 respectively at 52nd week after drug withdraw. White blood cells began to be elevated at 4th week and were restored to the normal levels at 8th week in the treatment group, while the count in the control was lower than the normal value even at 52nd week of the treatment with the average of (3.45±1. 18)×109/L. Conclusion Alpha-interferon combined with mannan peptide therapy is effective for patients with HBeAg-positive CHB, which may restore the declined peripheral WBC counts induced by interferon and improve the compliance.

Objective To summarize the experiences of Nuss procedure for complicated pectus excavatum.Methods From August 2006 to July 2011,443 patients with pectus excavatum received Nuss operation.Among them,95 patients with complicated pectus excavatum (CT Haller index ＞ 6) received modified technique of Nuss procedure,including multi-bar technique,oblique placed bar,double arc bar technique; fix with wire ; osteotomies ; auxiliary small incision; thoracoscopy placed by different lateral.Results All operations were successfully accomplished without severe complications.The mean operative time was (90.13 ± 39.12 ) minutes and the mean volume of blood loss was (45.41 ± 19.23 )ml.The mean hospital stay was (7.21 ± 2.87 ) days.All patients have been satisfied with their surgical correction.Therapeutic results evaluation was excellent in 75.8％ of patients,good in 16.8％.Conclusion Multi modified technique of Nuss procedure can lead to a satisfactory outcome for complicated pectus excavatum patients.

Hypoxia-induced 15-hydroxyeicosatetraenoic acid (15-HETE) is an essential mediator to constrict pulmonary arteries (PA). The signaling pathway involved in 15-HETE-induced PA vasoconstriction remains obscure. The aim of the present study was to test the hypothesis that hypoxic PA constriction induced by 15-HETE was possibly regulated by the extracellular signal-regulated kinase-1/2 (ERK1/2) pathway. PA ring tension measurement, Western blot and immunocytochemistry were used in the study to determine the possible role of ERK1/2 in 15-HETE-induced PA vasoconstriction. The organ bath for PA rings tension study was employed. Adult male Wistar rats were raised in hypoxic environment with fractional inspired oxygen (FIO2, 0.12) for 9 d. PA 1~1.5 mm in diameter were dissected and cut into 3 mm long rings for tension study. ERK1/2 up-stream kinase (MEK) inhibitor PD98059, which blocks the activation of ERK1/2, was used. The results showed that pretreatment of PD98059 significantly blunted 15-HETE-induced PA vasoconstrictions in the rings from hypoxic rat. Moreover, in endothelium-denuded rings, PD98059 also significantly attenuated 15-HETE-induced vasoconstriction. Phosphorylation of ERK1/2 in pulmonary arterial smooth muscle cells (PASMCs) of rat was enhanced evidently when stimulated by 15-HETE. Thus, the data suggest that ERK1/2 signaling pathway is involved in 15-HETE-induced hypoxic pulmonary vasoconstriction.
Animals ; Flavonoids ; pharmacology ; Hydroxyeicosatetraenoic Acids ; antagonists & inhibitors ; pharmacology ; Hypoxia ; physiopathology ; MAP Kinase Signaling System ; physiology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Pulmonary Artery ; cytology ; drug effects ; physiopathology ; Rats ; Rats, Wistar ; Vasoconstriction ; drug effects

Objective To determine the proportion of CD4+ CD25+ regulatory T (Treg) cells,mRNA expression of the forkhead box protein 3 (Foxp3) gene,and DNA methylation status of the Foxp3 promoter in peripheral CD4+ T cells from patients with Henoch-Sch(o)nlein purpura.Methods Totally,20 inpatients with Henoch-Sch(o)nlein purpura and 20 healthy controls were enrolled from Department of Dermatology,the Second Xiangya Hospital of Central South University between 2015 and 2016,and there were no significant differences in the gender and age between the two groups (both P ＞ 0.05).CD4+ T cells were isolated from the peripheral blood samples of these subjects.Real-time fluorescence-based quantitative PCR was performed to detect the mRNA expression of the Foxp3 gene,flow cytometry to determine the proportion of CD4 + CD25+ Treg cells,and sodium bisulfite sequencing PCR (BSP) to determine the DNA methylation status of the Foxp3 promoter.Statistical analysis was carried out with SPSS16.0 software by using two-sample t test for the comparison between the two groups,and linear correlation analysis for evaluating the correlations of the DNA methylation status of the Foxp3 promoter with clinical severity scores and the proportion of CD4+CD25+ Treg cells.Results Compared with the healthy control group,the Henoch-Sch(o)nlein purpura group showed significantly decreased mRNA expression of the Foxp3 gene in CD4+ T cells (0.380 ± 0.226 vs.1,t =9.503,P ＜ 0.01),proportion of CD4+CD25+ Treg cells (1.668％ ± 0.959％ vs.2.741％ ± 1.131％,t =2.552,P ＜ 0.05),but significantly increased DNA methylation status of the Foxp3 promoter (0.712 ± 0.164 vs.0.453 ± 0.147,t =3.610,P ＜ 0.01).In the Henoch-Sch(o)nlein purpura group,the DNA methylation status of the Foxp3 promoter was negatively correlated with the percentage of CD4+CD25+ Treg cells (r =-0.490,P ＜ 0.05),but positively correlated with the clinical severity scores (r =0.486,P ＜ 0.05).The DNA methylation level of the Foxp3 promoter was significantly higher in the patients with renal impairment than in those without renal impairment (P ＜0.05).Conclusion The patients with Henoch-Sch(o)nlein purpura showed increased DNA methylation status of the Foxp3 promoter in CD4+ T cells,decreased mRNA expression of the Foxp3 gene and proportion of CD4+CD25+ Treg cells,which may be related to the occurrence of Henoch-Sch(o)nlein purpura,and affect disease development and prognosis.