OBJECTIVE:To study the mechanism of histone deacetylase inhibitor RGFP109 in reversing resistance of glioma U251 cells. METHODS:TR/U251 cells resistance to temozolomide(TMZ)was extrablished. The test was divided into normal con-trol group,TMZ group(40 μmol/L)and TMZ(40 μmol/L)+RGFP109(0-120 μmol/L)different concentrations groups. After 24 h of adding into related medicines,CCK-8 was used to detect the cell survival rate and calculate the half inhibitory concentration (IC50). TUNEL and Annexin V/PI were used to detect the cell apoptosis in normal control group,TMZ group and TMZ+RGFP109 (42μmol/L)group. Immunoblotting was used to detect the O6-methyl guanine-DNA methyltransferase(MGMT),Survivin,B lym-phoma 2(Bcl-2),B lymphoma xL(Bcl-xL)protein expression;and gel migration test was used to detect the p65 acetylation level and its binding capacity with κB-DNA. RESULTS:Compared with normal control group,cell survival rate in TMZ+RGFP109 dif-ferent concentrations groups was obviously decreased (P<0.05),showing a concentration-dependent manner. When the RGFP109 concentration was 42 μmol/L,the sensitivity of TMZ to TR/U251 cells was the same with U251 cells. Compared with normal con-trol group,MGMT,Survivin,Bcl-2,Bcl-xL protein expressions in cells of TMZ groups were enhanced(P<0.01);p65 acetyla-tion level had no obvious changes,while the binding capacity of p65 and κB-DNA was strengthened (P<0.01). Compared with TMZ group,MGMT,Survivin,Bcl-2,Bcl-xL protein expressions in cells of TMZ groups were weakened(P<0.01);p65 acetyla-tion level was enhanced (P<0.01);and the binding capacity of p65 and κB-DNA was weakened (P<0.01). CONCLUSIONS:RGFP109 can reverse the resistance of U251 cells to TMZ by down-regulating the anti-apoptotic protein expressions adjusted by transcription factorκB(NF-κB)and weakening the binding of p65 andκB-DNA.

Objective To explore the effect on metabolism of glucose and lipids, potential toxicity mechanism and possible biomarker candidates by analyzing urine metabonome changes of rats after oral administration of polychlorinated biphenyl congeners( PCBs) and high fat diet alone or in combination.Methods Male SD rats were divided randomly into control group,high fat diet group, PCBs group and combination group of PCBs and high fat diet.Urine samples were collected after 6-week treatment, 1 H NMR spectra were performed and analyzed by principal component analysis ( PCA) . Results The PCA scores plot of urine 1 H NMR data showed that the combined group could be easily distinguished from the other three groups, suggesting great difference in metabolism.The loading plot of the PCA revealed significant increase in the levels of lactate, glucose, creatine, 2-hydroxy-isovaleric acid and reduction in the levels of citrate, succinate, taurine, hippurate and trimethylamine oxide ( TMAO) in the combined exposure group after six-week exposure.Conclusion The altered levels of metabolites induced by combined exposure of PCBs and high fat diet may be related to the injury to mitochondrial function, reduction of energy metabolism in tricarboxylic acid cycle (TAC).These effects possibly lead to perturbations in the metabolism of glucose, lipid and amino acids.The altered metabolites may be considered biomarker candidates of toxicity induced by exposure to PCBs and high fat diet.

The rat model of dysmenorrhea, pelvic inflammation disease (PID), and uterine myoma were established to observe the effect ofGui-Zhi Fu-Ling Capsule (GZFLC) with the changes of main components. The molecular imprinted technology was used to quantitatively knock out main components in GZFLC (the knocked-out ratios were 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%). And the observation was made on the writhing times, latency, and effects on ET-1 and PGF2α of uterine tissues in the pitocin-induced dysmenorrhea rat model. Effects on TNF-α and IL-2 were observed in PID rat model induced by mixed bacteria plus mechanical damages. Effects on the weight and index of uterus, levels of E2 and P in serum were detected on the uterine myoma rat model induced by estrogen-burden. The results showed that the GZFLC efficacy was decreased with gradient knockout of 15 main components. When 10%-40% was knocked out, there was still significant difference compared to the model group (P < 0.01). However, when 50%-90% was knocked out, there was no significant difference compared to the model group (P > 0.05). Meanwhile, the GZFLC efficacy was decreased significantly compared to the GZFLC group (P <0.05, orP < 0.01). It was concluded that 15 chemical compounds, which included gallic acid, paeoniflorin, paeonol, and etc., may be the main material basis of GZFLC. Meanwhile, it provided a useful reference for the quality control of main components in GZFLC.

OBJECTIVETo establish a simple extraction, isolation and purification method for ginkgolide B from ginkgo leaf.

METHODThe optimum conditions of extraction, isolation and purification were studied by taking the transfer rate of ginkgolide B as index.

RESULTGinkgo leaf was extracted with 70% ethanol for three times, the extracts were concentrated to remove ethanol and diluted by water till the crude drug density reached 0.1 g x mL(-1). The dilution was adsorbed with HPD-450 macroporous resin. The impurities were eluted with 20% ethanol and ginkgolide B was eluted with 80% ethanol. Then the 80% ethanol eluant was concentrated and crystallized. Finally the crude crystals were recrystallized with isopropanol. The purity of the ginkgolide B recrystallization was 95%.

CONCLUSIONThe process was stable and easy to operate, which was suited to industrialized production.

Chemical Fractionation ; methods ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Ginkgo biloba ; chemistry ; Ginkgolides ; analysis ; isolation & purification ; Lactones ; analysis ; isolation & purification ; Plant Leaves ; chemistry

OBJECTIVE To investigate the role of gonadotropin-releasing hormone (GnRH) in es-tradiol(E2 ) induced advance of puberty onset in rats. METHODS Postnatal day 18 SD rats were given a daily intragastric administration of corn oil or E2(50 μg.kg-1 ) for consecutive 5 d. The day of vaginal opening (VO), pathological changes in ovary and protein expression levels of GnRH, G protein-coupled receptor 54 ( GPR54) and phospholipase C ( PLC) in hypothalamus were observed. RESULTS As compared to corn oil controll group, VO was advanced by about 12.2 d, corpus luteum was observed in the ovary section, and the protein expression levels of GnRH,GPR54 and PLC in hypothalamus were significantly increased by 47%, 55% and 56% in E2 group, respectively. CONCLUSION E2 induced onset of puberty advance may be closely related to regulation of the expression of GnRH, GPR54 and PLC in hypothalamus.

OBJECTIVETo develop a new method to rapidly determine and identify Guizhi Fuling capsule by portable acousto-optic tunable filter-near infrared spectroscopy.

METHODThe qualitative model was set up using principal component analysis. The correlation models between the NIR spectra and the reference values of five major constituents were obtained with partial least squares method.

RESULTThe identifying model accurately identified Guizhi Fuling capsule, and quantitative analytical models could precisely predicted the content of ellagic acid, baicalin, benzoylpaenoniflorin, cinnamaldehyde, and paeonol. The correlation coefficients of the calibration models were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, the validation set coefficients of the calibration were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, and the RMSEP were 1.138%, 3.014%, 0.751%, 0.625%, 3.455%, 1.363%, respectively. The results of external validation showed no significant difference between the predictive and the determining values by t-test.

CONCLUSIONThe method is accurate, rapid and non-destructive, and can be used for determining and identifying Guizhi Fuling capsule.

Acetophenones ; analysis ; Acrolein ; analogs & derivatives ; analysis ; Calibration ; Capsules ; chemistry ; Drug Evaluation ; methods ; Drugs, Chinese Herbal ; analysis ; Ellagic Acid ; analysis ; Flavonoids ; analysis ; Least-Squares Analysis ; Models, Chemical ; Principal Component Analysis ; methods ; Spectroscopy, Near-Infrared ; methods

OBJECTIVEThe aim of this study was to examine the effect of low dose heavy ion irradiation on the subset percentage and expression of cytokines of peripheral blood lymphocytes(PBL) in patients with pancreatic cancer.

METHODSPBL from 21 patients with pancreatic cancer were divided into three groups: sham, X-ray and ¹²C⁶⁺ irradiation groups, and the cell responses were measured at 24 hours after radiation exposure. The percentages of T and NK cell subsets were detected by flow cytometry. The mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real-time quantitative RT-PCR (qRT-PCR). The cytokine protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA).

RESULTSThe percentage of T lymphocyte subsets was significantly increased at 24 hours after exposure to low dose radiation, and the effect was more pronounced in the group receiving 0.05 Gy ¹²C⁶⁺ ion irradiation than that in the group receiving X-ray irradiation [CD3⁺ T cells: (67.15 ± 4.36)% vs. (60.81 ± 8.35)%; CD3⁺ CD4⁺ T cells: (19.02 ± 2.35)% vs. (17.21 ± 2.86)%; CD3⁺ CD8⁺ T cells: (46.59 ± 6.07)% vs. (41.18 ± 6.35)%. (P < 0.05 for all)]. However, there were no significant changes in the CD3⁺ CD4⁺/CD3⁺ CD8⁺ ratio (0.67 for sham, 0.65 for X-ray, and 0.68 for ¹²C⁶⁺ groups) and percentage of NK cell subsets (P > 0.05 for all). Expression levels of IFN-γ mRNA (cycle threshold/CT value was 23.35 ± 3.16 for ¹²C⁶⁺, CT value was 27.25 ± 2.15 for X-ray) and IL-2 (CT value was 24.19 ± 3.56 for ¹²C⁶⁺, CT value was 27.85 ± 4.08 for X-ray) in PBL, and their protein levels in the supernatant were significantly increased at 24 hours after exposure to the low dose radiation (P < 0.05). The effects were more pronounced in the group receiving 0.05 Gy ¹²C⁶⁺ ion irradiation than that in the group receiving X-ray irradiation. However, there was no significant change in the TNF-α production of PBL.

CONCLUSIONSLow dose irradiation may alleviate the immune suppression caused by tumor burden and that the effect is more pronounced for 0.05 Gy high linear energy transfer (LET) ¹²C⁶⁺ irradiation. The percentage of T cell subsets and cytokines production could be used as sensitive indicators of acute response to low dose irradiation.

CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Cytokines ; metabolism ; Dose-Response Relationship, Radiation ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Heavy Ions ; Humans ; Interleukin-2 ; metabolism ; Killer Cells, Natural ; Lymphocytes ; metabolism ; radiation effects ; Pancreatic Neoplasms ; metabolism ; radiotherapy ; Tumor Necrosis Factor-alpha ; metabolism

Objective:To compare the effects of high-power and conventional power atrial fibrillation ablation on intraoperative acute pulmonary vein isolation, postoperative troponin levels, and atrial fibrillation recurrence.Methods:A retrospective selection was conducted on 105 patients with paroxysmal atrial fibrillation admitted to the Baoding NO.1 Central Hospital from January 2017 to December 2020. According to different treatment methods, they were divided into a high-power ablation group of 52 cases and a conventional power ablation group of 53 cases. The intraoperative rate of single circle acute pulmonary vein isolation, the recovery of electrical conduction after acute pulmonary vein isolation, and the location and number of points that need to be added were compared between the two groups; At the same time, two groups were compared in terms of surgical time, ablation time, surgical radiation exposure time and radiation dose, intraoperative complications postoperative cardiac troponin levels at 12 hours, and recurrence of atrial fibrillation within 1 year after ablation.Results:The intraoperative single loop pulmonary vein isolation rate and postoperative troponin levels in the high-power atrial fibrillation ablation group were higher than those in the conventional atrial fibrillation ablation group (all P<0.05). The surgical time, ablation time, and the number of sites and points that need to be added during surgery were less than those in the conventional atrial fibrillation ablation group (all P<0.05). There was no statistically significant difference in the incidence of intraoperative complications and postoperative atrial fibrillation recurrence between the two groups (all P>0.05). Conclusions:High power atrial fibrillation ablation has a higher single loop acute pulmonary vein isolation rate, fewer patch sites and points, shorter surgical time, and greater ablation damage compared to conventional ablation, and the clinical efficacy of the two groups is similar after surgery.

Objective:To explore the risk factors and time distribution of renal relapse in patients with lupus nephritis (LN).Methods:Clinical, pathological characteristics and long-term outcomes of LN patients who were diagnosed and followed in Jinling Hospital from January 2004 to December 2008 were retrospectively analyzed. The patients were divided into relapse group and non-relapse group. The differences of clinical pathological characteristics between the two groups were compared. The multivariate Cox proportion risk model was used to analyze the risk factors affecting renal relapse in LN. The risk factors and time distribution of renal relapse were analyzed with annual relapse risk-time curve.Results:A total of 814 patients with LN were included in the study, with 419 cases (51.5%) of complete remission and 395 cases (48.5%) of partial remission. The age was (30.24±10.90) years old, and there were 112 males (13.8%). There were 367 patients suffering renal relapse. The time of first renal relapse was (3.21±2.70) years. The results of multivariate Cox regression showed that age ( HR=0.976, 95% CI 0.966-0.986, P<0.001), renal pathological activity index (AI) score ( HR=1.039, 95% CI 1.013-1.065, P=0.003), remission status after induction treatment ( HR=0.671, 95% CI 0.504-0.894, P=0.006), 24 h urinary protein quantitation ( HR=1.297, 95% CI 1.115-1.508, P=0.001), anti-double strand DNA antibody (A-dsDNA, HR=1.450, 95% CI 1.139-1.846, P=0.003) and complement C3 ( HR=0.223, 95% CI 0.128-0.389, P<0.001) were correlated with increasing risk of renal relapse in LN. The annual relapse risk profile was unimodal, with a peak period of the second year after maintenance treatment. Similar patterns of relapse were presented in subgroup analysis. Conclusions:Age, renal pathological AI score, remission status after induction therapy, 24 h urine protein, A-dsDNA and blood complement C3 are the influencing factors for relapse of LN patients. The peak period of renal relapse in patients with LN is in the second year of maintenance therapy.

ObjectiveTo explore the efficacy and mechanism of the alcohol extract DH50 of Angelicae Pubescentis Radix in treating gouty arthritis induced by monosodium urate （MSU） crystals in vivo and in vitro. MethodFifty male SD rats were randomly assigned into five groups （n=10）： a normal group， a model group， a dexamethasone （DXMS， 0.07 mg·kg^-1） group， and low- （DH50-D， 9 mg·kg^-1） and high-dose （DH50-G， 18 mg·kg^-1） DH50 groups. The rats in the normal group and model group were administrated with the same amount of pure water. On day 5， the gouty arthritis model was established by injecting MSU into the right ankle joint of rats. The toe volume and joint inflammation index were measured 4， 8， 24， and 48 h after modeling. The pathological changes of the synovial tissue were detected by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， and IL-6 in the synovial tissue. Western blot was employed to measure the protein levels of NOD-like receptor protein 3 （NLRP3）， cysteine-aspartic protease-1 （Caspase-1）， apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain （ASC）， IL-1β， and cyclooxygenase-2 （COX-2） in the synovial tissue. Furthermore， the cell inflammation model was established with RAW264.7 cells stimulated with MSU （75 mg·L^-1）. The cell experiments were carried out with 6 groups： a normal group， a model group， a positive drug （DXMS， 100 μmol·L^-1） group， and low- （DH50-D， 25 mg·L^-1）， medium- （DH50-Z， 50 mg·L^-1）， and high-dose （DH50-G， 100 mg·L^-1） DH50 groups. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the cell viability， ELISA to determine the content of TNF-α in the supernatant of cell culture， and Western blot to determine the protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2. ResultCompared with the normal group， the rat model group showed increased toe swelling degree and joint inflammatory index （P<0.01）， serious infiltration of the synovium， elevated levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， ASC， IL-1β， and COX-2 （P<0.05， P<0.01）. Compared with the rat model group， low- and high-dose DH50 mitigated the toe swelling degree， decreased the joint inflammatory index， alleviated the inflammatory infiltration， lowered the levels of inflammatory cytokines in the tissue homogenate （P<0.01）， and down-regulated the expression of related proteins （P<0.05， P<0.01）. Compared with the normal group， the cell model group showed elevated level of TNF-α in the supernatant （P<0.01） and up-regulated protein levels of NLRP3， cleaved Caspase-1， IL-1β， TNF-α， and COX-2 （P<0.05）. Compared with the model group， low， medium， and high doses of DH50 lowered the level of TNF-α in the supernatant of cell culture in a dose-dependent manner and down-regulated the expression of related proteins （P<0.05， P<0.01）. ConclusionDH50 can mitigate gouty arthritis both in vitro and in vivo by inhibiting the activation of NLRP3 inflammasomes and the production of inflammatory cytokines.