AIM: To establish HPLC fingerprint of Ramulus cinnamomi.This fingerprint was expected to be standards of quality control and identification of the Chinese crude drug. METHODS: The HPLC method was used,chromatography condition were Kromasil C_(18)(250 mm?4.6 mm,5?m) column with gradual mobile phase of acetonitrile-0.5% acetic acid,UV detection wavelength at 280 nm and column temperature at 25(?C) with the flow rate of 1 ml?min~(-1). RESULTS: The retention time of Cinnamaldehyder,Cinnamic acid,o-Methoxycinnamic acid and Coumarin in Ramulus cinnamomi were determined.The fingerprint showed high similitude in samples from different regions. CONCLUSION: The HPLC fingerprint of Ramulus cinnamomi.can be used as a standard of quality control and identification of the Chinese crude drug.

OBJECTIVE:To determine the solubility and apparent oil and water partition coefficient (lg P) of saponin H1,and provide reference for dosage form design and druggability research of saponin. METHODS:HPLC-ELSD was conducted to deter-mine the equilibrium solubility of saponin H1 in different organic solvents and pH buffer solutions;shaking flask was applied to de-termine lg P value of saponin H1. RESULTS:The equilibrium solubility of saponin H1 in water,methanol and ethanol at 25 ℃ was 0.09175 g/L,96.51 g/L and 46.89 g/L,respectively. The solubility increased apparently at high pH within pH range at 7.6-10.0;the lg P value was between 0.695-0.773 in buffers within pH range at 6.0-8.0. CONCLUSIONS:Saponin H1 and the oleanolic acid type saponins belong to low solubility, low transmission components, so it is not suitable for use as a conventional oral formulation is developed.

Objective To explore the reasons for lateral and basal incomplete resection of precancerous lesions or cancer from upper digestive tract by endoscopic submucosal dissection (ESD).Methods Data of 295 patients undergoing ESD for upper gastrointestinal precancerous lesions or cancer from November 2006 to October 2011 were collected,and reasons of basal or lateral incomplete resectin confirmed by postESD pathology were analyzed.Results The total incomplete resection rate after ESD was 3.05％ ( 9/295 ).Among 95 cases of esophageal ESD,there was 1 case of lateral margin incomplete resection because of the retraction of normal tissue after dissection.Among 200 cases of gastric ESD,there were 5 cases of lateral margin incomplete resection,in which 2 cases were signet ring carcinoma with submucosal infiltration and spreading,2 were due to retraction of normal tissue after dissection,and 1 was due to inaccurate judgment on cancer demarcation.There were 3 cases of basal incomplete resectin in gastric ESD,which was caused by incorrecte invasion depth estimation before ESD.Conclusion The rate of basal or lateral incomplete resection in upper gastrointestinal ESD was low,which is related to pathological type,ESD procedure and estimation of invasion depth before ESD.

Objective To complete the synthesis of the natural product of crinumaquine .Methods 3 ,4-methylenedioxy-phenethylamine and 2 ,5-dimthoxyphenylacetic were taken as starting material ,and chemical reactions of condensation ,cycliza-tion ,reduction ,oxidation and other reactions were conducted .Results and Conclusion The optimum synthetic route was deter-minedunder which the final yield rate was 73% .

Objective:To compare the clinical efficacy, adverse reactions and prognosis of different radiotherapy schemes in the treatment of advanced esophageal squamous cell carcinoma.Methods:The clinical data of 60 patients with stage ⅣB esophageal squamous cell carcinoma who received radiotherapy in Rugao People′s Hospital of Jiangsu Province from January 2015 to January 2020 were retrospectively analyzed. According to the radiation doses, the patients were divided into standard dose group (total radiation dose <50.4 Gy) and high dose group (total radiation dose ≥50.4 Gy), with 30 patients in each group. The scores of dysphagia before and after treatment were analyzed by Wilcoxon signed-rank test. The radiotherapy effective rate, remission rate of dysphagia and incidence of adverse reactions were analyzed by χ2 test. Survival analysis was carried out by Kaplan-Meier and log-rank test was used to compare the prognosis of the two groups. Results:There were no significant differences in dysphagia scores between standard dose group and high dose group before and after radiotherapy ( Z=1.232, P=0.876; Z=1.506, P=0.278). The dysphagia symptoms were relieved after radiotherapy in all patients, and the dysphagia score was significantly higher than that before radiotherapy ( Z=6.347, P<0.001). The radiotherapy effective rates in the standard dose group and high dose group were 76.7% (23/30) and 83.3% (25/30) respectively, with no statistically significant difference ( χ2=0.417, P=0.519). The remission rates of dysphagia in the two groups were 80.0% (24/30) and 90.0% (27/30) respectively, with no statistically significant difference ( χ2=0.523, P=0.470). The incidences of adverse reactions in the two groups were 43.3% (13/30) and 83.3% (25/30) respectively, with a statistically significant difference ( χ2=10.335, P=0.001). The median overall survival in the standard dose group and high dose group were 11 months and 9 months respectively, with no statistically significant difference ( χ2=1.490, P=0.256). Conclusion:There are no statistical differences in short-time efficacy, symptom remission and long-term prognosis between the standard dose group and the high dose group in patients with advanced esophageal squamous cell carcinoma. However, the incidence of adverse reactions in patients receiving standard dose radiotherapy is low, which is worthy of clinical application.

OBJECTIVETo explore the role of transforming growth factor-β1 (TGF-β1) in intervertebral disc degeneration and its association with the pathological grading of disc degeneration.

METHODSNormal and degenerative intervertebral disc tissues were collected were classified into 5 grades of increasing degenerative changes. HE staining, immunohistochemistry, TUNEL staining and RT-PCR were used to detect the expression of TGF-β1 in the disc tissues.

RESULTSImmunohistochemistry and RT-PCR showed positive expressions of TGF-β1 and Bcl-2 in normal disc tissues, where Bax was expressed at have a trace level. In the degenerative disc tissues, TGF-β1 expression increased with the pathological grades; the expression levels of TGF-β1 showed significant differences between degenerative and normal tissues and between grade IV and grade I disc tissues (P<0.01).

CONCLUSIONTGF-β1 is an important factor participating in the disc degeneration and its expression level is closely related to the pathological grade of degenerative discs.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Degeneration ; classification ; metabolism ; pathology ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVE:To set up a method for determination of molecular weight and content of polysaccharide in Bletilla striata. METHODS:HPSEC-ELSD method was adopted. The determination was performed on TSK-GEL G4000 PWXL column with mobile phase consisted of pure water at the flow rate of 0.6 mL/min.The column temperature was 30 ℃,and sample size was 20 μL. Evaporative light scattering detector was used.The molecular weight and content of polysaccharide in 3 batches of B. striata were established. RESULTS:The linear range of weight average molecular weight of B. striata polysaccharide were 24.17-178.00 kD(R2=0.985 5). The linear range of BT07 content determination were 0.508 0-5.080 mg/mL(R2=0.998 4). The limits of detection and quantitation were 0.116 5 and 0.274 0 mg/mL. RSDs of precision,stability and reproducibility tests were all lower than 2%(n=6 or n=7).Average recoveries rate were 99.16%-100.20%(RSD=0.39%-0.64%,n=9).Average retention time of 3 batches of samples was 14.28 min(RSD=4.35%,n=3),and weight average molecular weight was 23.54 kD(RSD=3.78%, n=3). Polydispersity coefficient D,MW/Mn)ranged 1.463-1.578. Average content of sample was 93.4%(RSD=4.22%,n=3). CONCLUSIONS:HPSEC-ELSD method is simple,rapid,accurate and reliable,which can be used for simultaneous determination of B.striata polysaccharide.

OBJECTIVETo investigate the effects of CysLT receptor agonist leukotriene D4(LTD4) and antagonists on activation of microglia BV2 cells.

METHODSThe expression of CysLT1 and CysLT2 protein was determined by Western blotting and immunostaining in microglia BV2 cells. BV2 cells were pretreated with or without CysLT1 receptor selective antagonist montelukast, CysLT2 receptor selective antagonist HAMI 3379, or CysLT1/CysLT2 receptor dual antagonist BAY u9773 for 30 min, then the cells were treated with LTD4 for 24 h. Cell viability was detected by MTT reduction assay. Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR, respectively.

RESULTSIn BV2 cells, LTD4 did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner. LTD4 at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression (P<0.01). HAMI 3379 and BAY u9773 (100 nmol/L) further increased LTD4-induced phagocytosis; BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression, while HAMI 3379 had no effect on that.

CONCLUSIONLTD4 activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT1 receptor, LTD4 induces phagocytosis which might be negatively regulated by CysLT2 receptor in BV2 cells.

Acetates ; pharmacology ; Cell Line ; Cell Proliferation ; Cyclohexanecarboxylic Acids ; pharmacology ; Humans ; Interleukin-6 ; metabolism ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Microglia ; cytology ; metabolism ; Phagocytosis ; Phthalic Acids ; pharmacology ; Quinolines ; pharmacology ; Receptors, Leukotriene ; metabolism ; SRS-A ; analogs & derivatives ; pharmacology

Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.

OBJECTIVE： To establish the separation and purification technology of sanguinarine from the extract of Macleaya cordata with ion exchang resin. METHODS： The content of sanguinarine from the extract of M. cordata was determined by HPLC， with  Cosmosil C18-R-Ⅱ column （250 mm×4.6 mm，5 μm）， mobile phase of acetonitrile-0.2％ acetic acid solution （25 ∶ 75，V/V）， the flow rate of 1 mL/min， detection wavelength of 270 nm， column temperature of 30 ℃， and sample size of 20 μL. Static adsorption and desorption tests were carried out to compare the adsorption and desorption properties of 8 ion exchange resins for sanguinarine. The optimum concentration of sample solution， pH value and volume of sample were investigated by optimum ion exchange resin. APPS 10D liquid phase preparation system was used to investigate the dynamic elution conditions and obtain M. cordata refined extract solution. The refined purified product of M. cordata was obtained by desalination， elution on a reversed-phase （RP） C18 column and drying.  The purity of the purified product was analyzed by HPLC. The structure of the purified product was confirmed by HPLC， UV spectrophotometry， MS and NMR. RESULTS： CM-FF resin was screened for the separation and purification of sanguinarine from M. cordata extract. It was eluted with 20 mmol/L ammonium acetate solution 100 mL containing 20％ methanol and 0.25 mol/L sodium chloride. The optimal dynamic absorption condition included that the concentration of sample was 6.0 mg/mL at pH 5.0，and the loading amount was 25 mL； after desalination and refinement， for the eluted refined extract， the purified product with 97％ purity （purified yield  of 71％） was obtained， and its structure was confirmed to be sanguinarine. CONCLUSIONS： The optimal separation and purification technology by ion exchange resin is green， safe， efficient and easy to operate， which can be used for the separation and purification of sanguinarine from M. cordata extract and is suitable for industrial production.