The role of PPARγ in intermittent hypoxia-related human umbilical vein endothelial cell injury.

PURPOSE: Patients with obstructive sleep apnoea (OSA) have a high incidence of vascular endothelial injury. The most important pathophysiological feature of OSA is chronic intermittent hypoxia (CIH). This study aimed to investigate the mechanisms of CIH-related vascular endothelial injury. METHODS: IH exposure was applied to human umbilical vein endothelial cells (HUVECs). After modeling, cell viability, the expression levels of peroxisome proliferator activated receptor γ (PPARγ), apoptosis-associated proteins and mitochondrial division fusion proteins, and the levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were assessed via Cell Counting Kit-8 (CCK-8), western blotting, fluorescent microscope, and flow cytometry, respectively. Rosiglitazone (PPARγ agonist), tempo (the mitochondrial-specific antioxidant), and tempo combined with PPARγ interfering RNA were used to treat HUVECs, respectively. RESULTS: After IH exposure, cell viability and levels of MMP decreased, cell apoptosis and ROS levels increased, and the expression levels of PPARγ decreased. Both tempo and rosiglitazone pretreatment ameliorated cell apoptosis and improved cell viability. In addition, mitochondrial function became better after tempo pretreatment. PPARγ interference reversed the protective effects of tempo on IH-related mitochondrial function injury and cell injury. CONCLUSIONS: PPARγ regulated the apoptosis and cell viability of IH-treated HUVECs by altering mitochondrial function. This finding clarifies the mechanism of CIH-related vascular endothelial injury.

Comprehensive chemical study on different organs of cultivated and wild Sarcandra glabra using ultra-high performance liquid chromatography time-of-flight mass spectrometry (UHPLC-TOF-MS) / 中国天然药物

To illuminate the similarities and differences between wild and cultivated Sarcandra glabra (S. glabra), we performed a comprehensively study on 26 batches of cultivated S. glabra and 2 batches of wild S. glabra. Chemical constituents and distribution characteristics of roots, stems and leaves in both wild and cultivated S. glabra were investigated through UHPLC-TOF-MS method. The result revealed that there were significant differences between roots, stems and leaves in S. glabra. And the chemical contents in the root part were less or even absence than those in leaf and stem, which suggested the root organ could be excluded as medicine. Meanwhile, the chemical contents of stems and leaves in cultivated S. glabra was sightly higher than that of wild samples. Therefore, cultivated S. glabra may have a high potential for substitution of wild S. glabra without affecting its pharmaceutical properties. In summary, our study could provide important information to the molecular basis for quality control of S. glabra.

Clinical characteristics and drug sensitivity in children with invasive pneumococcal disease: a multicenter study / 中国当代儿科杂志

OBJECTIVE@#To study the clinical characteristics, drug sensitivity of isolated strains, and risk factors of drug resistance in children with invasive pneumococcal disease (IPD).@*METHODS@#The clinical characteristics and drug sensitivity of the isolated strains of 246 hospitalized children with IPD in nine grade A tertiary children's hospitals from January 2016 to June 2018 were analyzed.@*RESULTS@#Of the 246 children with IPD, there were 122 males and 124 females. Their ages ranged from 1 day to 14 years, and among them, 68 (27.6%) patients were less than 1 year old, 54 (22.0%) patients were 1 to 2 years old, 97 (39.4%) patients were 2 to 5 years old, and 27 (11.0%) patients were 5 to 14 years old. Pneumonia with sepsis was the most common infection type (58.5%, 144/246), followed by bloodstream infection without focus (19.9%, 49/246) and meningitis (15.0%, 37/246). Forty-nine (19.9%) patients had underlying diseases, and 160 (65.0%) had various risk factors for drug resistance. The isolated Streptococcus pneumoniae strains were 100% sensitive to vancomycin, linezolid, moxifloxacin, and levofloxacin, 90% sensitive to ertapenem, ofloxacin, and ceftriaxone, but had a low sensitivity to erythromycin (4.2%), clindamycin (7.9%), and tetracycline (6.3%).@*CONCLUSIONS@#IPD is more common in children under 5 years old, especially in those under 2 years old. Some children with IPD have underlying diseases, and most of the patients have various risk factors for drug resistance. Pneumonia with sepsis is the most common infection type. The isolated Streptococcus pneumoniae strains are highly sensitive to vancomycin, linezolid, moxifloxacin, levofloxacin, ertapenem, and ceftriaxone in children with IPD.

Investigation into perturbed nucleoside metabolism and cell cycle for elucidating the cytotoxicity effect of resveratrol on human lung adenocarcinoma epithelial cells / 中国天然药物

In an effort to understand the molecular events contributing to the cytotoxicity activity of resveratrol (RSV), we investigated its effects on human lung adenocarcinoma epithelial cell line A549 at different concentrations. Cellular nucleoside metabolic profiling was determined by an established liquid chromatography-mass spectrometry method in A549 cells. RSV resulted in significant decreases and imbalances of deoxyribonucleoside triphosphates (dNTPs) pools suppressing subsequent DNA synthesis. Meanwhile, RSV at high concentration caused significant cell cycle arrest at S phase, in which cells required the highest dNTPs supply than other phases for DNA replication. The inhibition of DNA synthesis thus blocked subsequent progression through S phase in A549 cells, which may partly contribute to the cytotoxicity effect of RSV. However, hydroxyurea (HU), an inhibitor of RNR activity, caused similar dNTPs perturbation but no S phase arrest, finally no cytotoxicity effect. Therefore, we believed that the dual effect of high concentration RSV, including S phase arrest and DNA synthesis inhibition, was required for its cytotoxicity effect on A549 cells. In summary, our results provided important clues to the molecular basis for the anticancer effect of RSV on epithelial cells.

[Relationship between Early Treatment Response and Prognosis in Children with Acute Lymphoblastic Leukemia].

OBJECTIVE: To analyze the relationship between the early treatment response and the pregnosis in children with acute lymphoblastic leukemia(ALL). METHODS: Two hundred and Seventy-eight ALL children diagnosed and treated in Hainan general hospital from March 2013 to March 2017 were collected. All ALL children received therapy with CCLg-ALL-2008 regimen. The 3 year event-free survival (EFS) rate of ALL children in different groups was analyzed in terms of 4 indexes including sensitivity response to prednison at day 8 (D8-SRP), bone marrow remission at day 15 (D15-BMR) and at day 33 (D33-BMR), and minimal residual disease at day 33 (D33-BMR), and minimal residual disease at day 33(D33-MRD). These 4 indexes and other indexes possibly affecting the prognosis of ALL children were enrolled in Cox regression model for analysis of independent factors affecting the prognosis of ALL children. RESULTS: The D8-SRP test showed that among 269 ALL children, 240(89.22%) cases displayed prednisone poor response (PPR); the 3-year EFS rate in predrisone good response(PGR) group was significantly higher than that in PPR group(P<0.05). The D15-BMR detection showed that among 262 ALL children, the bone marrow remission(BMR) as M1 was observed in 230 cases (87.79%), M2 in 20 cases (7.63%) and M3 in 9 cases (4.58%); the 3-year EFS rate showed as follows:M1 group >M2 group >M3 group(P<0.05). The D33-BMR detection showed that among 257 ALL children, the BMR as M1 was observed in 227 cases (88.33%), M2 in 21 cses(8.17%) and M3 in 9 caes (3.51%); the 3-year EFS rate in 3 groups showed as follows: M1 group >M2 group >M3 group(P<0.05). The D33-MRD detection showed that among 185 ALL children, MRD<10-10 was found in 128 cases (69.19%), MRD≥10-4-10-2 in 43 cases (23.24%), MRD ≥10-2 in 14 cases (7.57%); the 3-year EFS rate in 3 groups showed as follows: MRD <10-4 group > MRD≥ 10-4-10-2 group>MRD≥10-2 group. The Cox regression analysis showed that PPR in D8-SRP test, M2 and M3 in D15 and D33 BMR detection, and MRD≥10-2 in D33 MRD detection as well as T-ALL typing were independent risk factors affecting the prognosis of ALL children. CONCLUSION: The early treatment response can predict the prognosis of ALL children, which is an independent prognostic factor for ALL children.

Quantitation of camellianin A in HepG2 cells using a high performance liquid chromatography-electrospray ionization tandem mass spectrometric method / 中国天然药物

The present study was designed to develop a sensitive and selective high performance liquid chromatography-tandem mass spectrometric method for the determination of Camellianin A in HepG2 cells. The extraction of Camellianin A was achieved using 15% trichloroacetic acid and then separated on a C column interfaced with a triple quadrupole tandem mass spectrometer in multiple reaction monitoring mode. The mobile phase was consisted of methanol-water (0.1% formic acid) (55 : 45, V/V). The total run time was 5.0 min. The method was linear in the concentration range of 0.25-250.0 ng·mL. The lower limit of quantification was 0.25 ng·mL. The intra- and inter-day relative standard deviations of entire concentration range were less than 9.3%. The proposed HPLC-MS/MS method was successfully applied to detect the intracellular concentration of Camellianin A in HepG2 cells.

Quantitation of ligupurpurosides B and C in rat plasma using HPLC-MS/MS / 中国天然药物

The present study was designed to develop a sensitive and selective specific high performance liquid chromatography (HPLC)-tandem mass spectrometric method (MS/MS) for the determination of ligupurpurosides B and C in rat plasma. The samples were prepared after protein precipitation and analyzed by liquid chromatography equipped with a C18 column interfaced with a triple quadrupole tandem mass spectrometer using ESI as the ionization source in the negative ion mode. The mobile phase consisted of water (0.01 % formic acid)-methanol (57 : 43, V/V) at the flow rate of 0.3 mL·min(-1). The analytes and internal standard acteoside were both detected by use of multiple reaction monitoring mode. The total run time was 6.0 min. The method was linear in the concentration range of 2.5-500.0 ng·mL(-1) and the lower limit of quantifiation (LLOQ) was 2.5 ng·mL(-1). The intra-day and inter-day relative standard deviations across three validation runs over the entire concentration range were less than 9.8 %. The accuracy determined at three concentrations was within ± 6.1% in terms of relative error. In conclusion, this assay offers advantages in terms of expediency and suitability for the analysis of ligupurpuroside B and ligupurpuroside C in various biological fluids.

Cloning, sequence analysis, and prokaryotic expression of GrGPPS gene in Gentiana rigescens / 中草药

Objective: To obtain the indispensable key enzyme involved in the monoterpene biosynthesis, the geranyl diphosphate synthase gene GrGPPS was cloned from Gentiana rigescens, and its sequence analysis and prokaryotic expression were performed. Methods: According to the GrGPPS gene sequence of transcriptome of triennial G. rigescens, a pair of specific primers was designed, and the full length of cDNA sequences was obtained by RT-PCR. Then TA cloning, sequencing and sequence analysing were performed. Prokaryotic expression vector pGEX-T-1-GrGPPS was constructed and transformed into Escherichia coli Rosetta for expression under 37°C and induced by 1 mmol/L IPTG. Results: The GrGPPS cDNA had a length of 1107 bp coding for 369 amino acids. Sequence analysis showed that GrGPPS was the member of "short-chain prenyltransferases" super family. Results of phylogenic analysis showed that GrGPPS was at the same evolutionary branch with AmGPPS. The SDS-PAGE results displayed that the expressed proteins were consistent with the anticipated size. Conclusion: The GrGPPS gene is cloned from G. rigescens, and the stable prokaryotic expression system of pGEX-T-1-GrGPPS is constructed. This work will provide a foundation for further purification, structure, and functional research of GrGPPS protein.

HPLC method for the determination of bezafibrate in human plasma and application to a pharmacokinetic study of bezafibrate dispersible tablet.

A sensitive and selective high-performance liquid chromatographic-UV (HPLC-UV) method for the determination of bezafibrate in human plasma has been developed. Sample treatment was based on protein precipitation with a perchloric acid-methanol solution 10:90 (v/v). Analytical determination was carried out by HPLC with UV detection at 235 nm. Chromatographic separation was achieved on a C18 column by isocratic elution with acetonitrile-ammonium acetate aqueous solution (10 mmol/L; pH 4.0) (44:56, v/v) at a flow rate of 1.0 mL/min. The method was linear in the concentration range of 0.1-15.0 microg/mL. The lower limit of quantitation was 0.1 microg/mL. The intra-and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 6.96%. The accuracy determined at three concentrations (0.2, 2.0, and 10.0 microg/mL for bezafibrate) was within +/- 10.0% in terms of accuracy. The method was successfully applied for the evaluation of pharmacokinetic profiles of bezafibrate dispersible tablet in 20 healthy volunteers. The results show that AUC, C(max), and T(1/2) between the testing formulation and reference formulation have no significant difference (P > 0.05). Relative bioavailability was 105.0 +/- 15.7%.

Effect of compound Puerarin on the collage IV in streptozotocin-induced diabetic nephropathy rats / 中华预防医学杂志

<p><b>OBJECTIVE</b>To observe the effect of compound Puerarin on collagen IV of streptozotocin-induced diabetic rats.</p><p><b>METHODS</b>Diabetic nephropathy rats were induced by intraperitoneal injection of streptozotocin (STZ). Rats were allocated randomly to control group (10), diabetes model group (10), Vitamin C group (10), Puerarin group (10), vitamin C plus Puerarin group (10). The study period lasted for 12 weeks. During and after the treatment, the general state, blood glucose levels, glycosylated hemoglobin, blood urea nitrogen, serum collagen IV, blood urea nitrogen, serum creatinine, urinary albumin excretion rate of the 24-hour, and clearance rate of creatinine collagen IV protein were determined by immunohistochemistoche analysis as well as type the gene expression of collagen IV alpha 1 mRNA were determined by in situ hybridization analysis in the kidney tissue of different groups.</p><p><b>RESULTS</b>(1) Diabetes mellitus and renal function lesion occurred in the four groups. (2) Vitamin C and Puerarin could improve the general conditions of diabetic Rats, decrease blood urea nitrogen [(8.68 +/- 0.43), (7.98 +/- 0.47) and (5.76 +/- 0.82) micromol/L, serum creatinine [(74.68 +/- 8.20), (75.52 +/- 7.98) and (58.66 +/- 6.65) mmol/L], and urinary albumin excretion rate of the 24-hour [(18.40 +/- 0.37), (17.24 +/- 0.30) and (9.97 +/- 1.27) mg/24 h x 10(-3)]; increase clearance rate of creatinine [(0.59 +/- 0.21), (0.61 +/- 0.14) and (0.69 +/- 0.32) ml/min], the expression of collage IV absorbance [(111.56 +/- 14.61), (110.78 +/- 9.69) and (95.44 +/- 9.97) ] in the diabetic Rats were significantly inhibited at the same time.</p><p><b>CONCLUSION</b>The compound Puerarin might have some functions on preventing ren by inhibiting expression of type IV collagen.</p>

Liquid chromatography-mass spectrometry method for the determination of venlafaxine in human plasma and application to a pharmacokinetic study.

A sensitive and selective liquid chromatographic-mass spectrometric (LC-MS) method for the determination of venlafaxine in human plasma has been developed. Samples were prepared using liquid-liquid extraction and analyzed on a C(18) column interfaced with a triple quadrupole mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase was methanol-water containing 10 mmol/L ammonium acetate, pH 7.9 adjusted with aqueous ammonia (80:20, v/v) at the flow rate of 1.0 mL/min. The analyte and internal standard clozapine were both detected by use of selected ion monitoring mode. The method was linear in the concentration range of 1.0-200.0 ng/mL. The lower limit of quantification (LLOQ) was 1.0 ng/mL. The intra- and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 10.1%. The accuracy determined at three concentrations (5.0, 50.0 and 150.0 ng/mL for venlafaxine) was within +/-10.0% in terms of relative error (RE). The method was successfully applied for the evaluation of pharmacokinetic profiles of venlafaxine capsule in 20 healthy volunteers. The results show AUC, T(max), C(max) and T(1/2) between the testing formulation and reference formulation have no significant difference (p > 0.05). Relative bioavailability was 103.4 +/- 14.1%.

Experimental study on treatment of minimal change nephropathy with Lespedeza michx / 中国中药杂志

<p><b>OBJECTIVE</b>To study the therapeutic effect of Lespedeza Michx in experimental rat model of minimal change nephropathy (MCN).</p><p><b>METHOD</b>The rat MCN model was reproduced by tail-intravenous injection of Adriamycin. The treatment effect of Lespedeza Michx was determined by the detection of urine protein collection for 24 hours, malondialdehyde (MDA), superoxide dismutase (SOD), ATCO, IL-8, TNF-alpha in serum, and renal histopathological changes were observed by microscope.</p><p><b>RESULT</b>Lespedeza Michx could decrease the contents of urine proteinuria, MDA, IL-8, and TNF-alpha in serum, increase SOD level and improved the pathological change of glomeruli.</p><p><b>CONCLUSION</b>Lespedeza Michx was effective on MCN.</p>

Effects of 3-substituted aryl oxindole(PH II-7) on cell cycle of tumor cells / 药学学报

<p><b>AIM</b>To study the antitumor mechanism of 3-substituted aryl oxindole (PH II-7) and determine its effects on cell cycle distribution of tumor cells.</p><p><b>METHODS</b>The cell cycle distributions were determined with FACS. The cell cycle regulation-related proteins of K562 lysates were analyzed with Western Blot. The inhibition of PH II-7 on DNA synthesis of tumor cells were estimated though 3H-thymidine incorporation and the tyrosine kinase activity of EGFR of A431 lysates was measured with ELISA.</p><p><b>RESULTS</b>PH II-7 effected cell cycle distribution of several tumor cells, including multidrug resistant tumor cell lines, and accumulation of cells in the G0-G1 stages was observed. The cell cycle regulation-related proteins CDK2, Rb and c-myc were inhibited by PH II-7 in a dose dependent manner, whereas the expression of CyclinE was increased after exposure to PH II-7. Furthermore, PH II-7 2.0 mg.L-1 was shown to inhibit the incorporation of 3H-thymidine into DNA, and 21.89%-41.29% of the PTK activity of EGFR in A431 lysates was inhibited by PH II-7 2-8 mg.L-1 in a dose-dependant manner.</p><p><b>CONCLUSION</b>PH II-7, a new anti-tumor agent, blocks the transition of cell cycle of tumor cells from G1 to S phase by inhibition CDK2.</p>

Effects of yishenjiangyafang on blood pressure and protecting renal function in RPH rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects and mechanism of Yishenjiangyafang, a prescription of Chinese traditional herbs used for renal hypertension, on blood pressure and protecting renal function of RPH rats.</p><p><b>METHOD</b>The 5/6 kidney of rat was resected to set up the RPH rat model. Blood pressure, Cr(creatinine), Ccr(creatinine clearance) and BUN(urea nitrogen) were measured dynamically. After eight weeks treatment, plasma content of PAR A II TXB2, 6-keto-PGF1 were measured. At same time, The change of renal pathology was observed.</p><p><b>RESULT</b>Yishenjiangyafang could reduce blood pressure Cr, but had no effect on BUN of RPH rat. The indexes of PAR, A II of each different dosage group of Yishenjiangyafang were decreased. At the same time, it reduced plasma content of TXB2, and increased 6-keto-PGF1 alpha. Glomerulosclerosis and atrophy of renal tubule in Yishenjiangyafang group RPH rats were better than those of the contrast group and the Capten group.</p><p><b>CONCLUSION</b>Yishenjiangyafang can reduce blood pressure of RPH rats and has protective effects on its kidney. Yishenjiangyafang can perform its effects of reducing blood pressure and protecting kidney by influencing the RAS of RPH rats.</p>

Blastic natural killer cell leukemia--one case report and review of literature / 中华血液学杂志

<p><b>OBJECTIVE</b>To report a case of blastic natural killer cell leukemia with an aggressive clinical course.</p><p><b>METHODS</b>The characteristics of blastic NK cell leukemia and its treatment were discussed with review of literatures.</p><p><b>RESULTS</b>After combination chemotherapy and spinal cord segmental radiotherapy, the patient entered hematological remission, but the extramedullary lesion remained unchanged.</p><p><b>CONCLUSION</b>Blastic NK cell leukemia has an aggressive clinical course with poor response to treatment and unfavorable prognosis.</p>