OBJECTIVE: To optimize the prescription and preparation process of garcinia glycoside bilayer tablets and study the drug release mechanism in vitro. METHODS: Solid dispersions were prepared by using water soluble carrier material polyethylene glycol (PEG) and then used to prepare the rapid release layer. Hydroxypropylmethyl cellulose (HPMC) was used as main sustained release material to prepare the sustained release layer. The formulations of rapid release layer and sustained release layer were optimized by using univariate analyses. DDslover software was used to fitting drug release curve. RESULTS: The content of the garcinia glycoside in the bilayer tablets was 98.5% and its release rate was consistent with the rule of release in vitro. The release behavior of the sustained release layer in vitro followed Higuchi equation (r=0.998 3). The main mechanism of drug release was the synergistic action of diffusion and dissolution. CONCLUSION: Garcinia glycoside bilayer tablets prepared by this method can achieve the goal of both rapid-acting and long-acting.

Xanthogranulomatous pyelonephritis (XGP) is an unusual form of chronic pyelonephritis in which the renal parenchyma is destroyed and replaced by lipid-laden foamy macrophages. It usually affects middle-aged women with a history of recurrent urinary tract infection, diabetes, or kidney stones. The inflammatory process is usually diffuse and can extend beyond the kidney. The rare focal forms may simulate primary renal tumours. The preoperative imaging diagnosis may be difficult. We reported five cases of XGP, The findings of ours were recorded including kidney size, shape, contour, the echogenecity of the renal parenchyma, the internal echoes of the dilate collecting system, the presence of perinephric fluid accumulation and obstruction. One of the 5 cases was a male patient, and the other four were female, with a mean age of 53 years. He affected kidneys of the 5 cases swelled in different degrees, and one of them was found with line-like anechoic fluid. Among the 5 cases, one kidney appeared as diffusely reducing of the parenchyma echogenicity, multiple hypoechoic areas, disappearance of corticomedullary differentiation and multiple hyperecho with shadow. A round cystic anechoic lesion was found in one kidney, with internal punctate echo and peripheral fluid. Ultrasonographic finding of 1 case was extremely hypoechoic lesion on the left kidney, protruding from the outline of the kidney, with the partial renal capsule discontinuous, the less clear boundary, and a little blood flow in it. Ultrasonographic demonstration of 2 cases was mild dilatation of the collecting system with irregular wall thickening and internal hypoechogenicity, and 1 case was solid lesion with less clear boundary to the pelvic wall and a small amount of blood flow signal, the another 1 case was showed floccule without internal blood flow. Three cases were caused by chronic obstruction verified by operation, of which one was staghorn calculi, one was poorly differentiated squamous cell carcinoma in the middle part of the ureter, and one was inflammatory stricture of upper ureteral. Through analysis of the above five cases and review of related literature, we explored diagnoses and management of the patients with XGP. Xanthogranulomatous pyelonephritis (XGP) is a rare chronic variant of pyelonephritis characterized by destruction of the renal parenchyma. Combining ultrasonographic features of XGP with clinical recurrent urinary infection and chronic obstruction, XGP can be included in the differentiation. The diagnosis of XGP suspected by ultrasound can be clarified by CT, MRI, contrast-enhanced ultrasound.
Female ; Humans ; Kidney/pathology* ; Male ; Middle Aged ; Pyelonephritis/surgery* ; Pyelonephritis, Xanthogranulomatous/surgery* ; Ultrasonography

This study was conducted to investigate the chemical constituents in the root of Dysosma versipellis (Hance) M. Cheng. The constituents were isolated by silica gel, lichroprep RP-C₁₈ and pharmadex LH-20 column chromatography and the IR, MS, NMR, 2D-NMR spectroscopic analysis were employed for the structural elucidation. Ten compounds were isolated from the 95% ethanol extract of Dysosma versipellis, their structures were elucidated as dysoverine D (1), dysoverine F (2), dysoverine A (3), podoverine A (4), α-peltatin (5), rutin (6), kaempferol-3-O-β-D-glucopyranoside (7), quercetin-3-O-β-D-glucopyranoside (8), kaempferol (9) and quercetin (10). Compound 2 is a new compound, and compounds 1 and 3-6 were isolated from this plant fo r the first time.

OBJECTIVE：To establish the metho d for the concentration det ermination of foretinib derivative LWK- 126 in liver microsomes，and to study its metabolism stability in liver microsomes of rats ，Beagle dogs and human. METHODS ：In the in vitro incubation system of liver microsomes ，LWK-126 was dissolved in liver microsomal incubation systems of rats ，Beagle dog and human initiated by reduced nicotinamide adenine dinucleotide phosphate solution. After incubation in water at 37 ℃ for 0，5，10，20， 30 and 60 min，the reaction was terminated with acetonitrile containing internal standard （1 μg/mL tolbutamide）. UPLC-MS/MS method was applied to determine the concentration of LWK- 126 in the incubation systems. The determination was performed on Waters BEH C 18 column with mobile phase consisted of water （containing 0.1％ formic acid ）-acetonitrile（containing 0.1％ formic acid）by gradient elution at the flow rate of 0.4 mL/min. The column temperature was 30 ℃，and the sample size was 2 μL. The mass spectral analysis was performed in a positive electrospray ionization mode ，and the full MS experiment was run with the selective reaction monitoring mode with a scanning range of m/z 50→1 200. Taking the concentration of LWK- 126 at 0 min as reference，the remaining percentage and the enzyme kinetic parameters were calculated. RESULTS ：The linear range of LWK- 126 was 0.05-15 μg/mL（R 2＝0.999 2）；the lower limit of quantification was 0.05 μg/mL，and the lowest detection limit was 0.01 μg/mL. The precision，accuracy，extraction recovery and matrix effect all met the analysis requirements of biological samples. The remaining percentage of LWK- 126 in liver microsomes of human ，rats and Beagle dogs for 60 min were （33.17±4.52）％，（3.14± 6.73）％，（1.38±5.85）％；t1/2 of them were 33.15，11.76，5.62 min；the clearance rates were 38.45，118.81，245.76 μL（/ min·mg）， respectively. CONCLUSIONS ：The method for the content ； determination of LWK- 126 in liver microsomes is established successfully. The order of metabolic stability of LWK- 126 in 〔2016〕4015） liver microsomes of different species is human ＞rats＞Beagle dogs.

To investigate the pharmacokinetic characteristics and metabolites of Src homology 2 region-containing protein tyrosine phosphatase 2 (SHP2) protein inhibitor in SD rats, a triazole quinolinone based derivative NC-55-122 was utilized. Firstly, rats were randomly divided into groups and given compound NC-55-122 intragastric and intravenous administration, respectively. Blood samples were collected at different time points. Taking carbmazepine as the internal standard, ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to determine the concentration of NC-55-122 in rats, and the methodology was verified. DAS 2.0 software was used to calculate the main pharmacokinetic parameters, and GraphPad Prism 8.0.1 software was used to plot the blood concentration-time curve. At the same time, UPLC-Q-TOF/MS was established to analyze plasma samples, and UNIFI metabolite prediction software was used to analyze metabolites after oral gavage and intravenous injection. The linear range of the mass concentration of compound NC-55-122 was from 1 ng·mL^-1 to 1 600 ng·mL^-1, and the linear relationship was good. The matrix effect, extraction recovery, precision, accuracy and stability were investigated in this linear range, which met the requirements of biological analysis. Secondly, the analysis of pharmacokinetic parameters showed that the oral bioavailability of the compound was low, with F%= 3.09%, indicating that the compound was absorbed slowly in vivo. Finally, five possible metabolites were deduced by analyzing the ion flow diagram and combining UNIFI software. The detection method established in this experiment is highly sensitive, specific, rapid and efficient, which is suitable for the determination of the blood concentration of compound NC-55-122 in rats and the analysis of metabolites, and lays a foundation for the structural modification and druggability evaluation of the later anti-tumor drug NC-55-122. All animal experiments were approved by the Experimental Animal Ethics Committee of Guizhou Medical University (approval number: 2200823).

The aim of this paper was to investigate the effect of SIRT1/TSC_2 signal axis on leukemia stem cell senescence induced by ginsenoside Rg_1. CD34~+CD38~- leukemia stem cells(CD34~+CD38~-LSCs) was isolated by magnetic cell sorting(MACS) and divided into two groups. The control group cells were routinely cultured, 40 μmol·L~(-1) ginsenoside Rg_1 was added to the control group for co-culture in Rg_1 group. The effect of Rg_l to induce CD34~+CD38~-LSCs senescence were evaluated by senescence-associated β-Galactosidase(SA-β-Gal) staining, cell cycle assay, CCK-8 and Colony-Assay. The expression of senescence associated SIRT1, TSC_2 mRNA and protein was examined by Real-time fluorescence quantitative PCR(FQ-PCR) and Western blot. The results showed that the CD34~+CD38~-LSCs could effectively be isolated by MACS, and the purity of CD34~+CD38~-LSCs is up to(95.86±3.04)%. Compared with the control group, the percentage of positive cells expressed SA-β-Gal in the Rg_1 group is increased, the senescence morphological changes were observed in the CD34~+CD38~-LSCs in the Rg_1 group. The proliferation inhibition rate and the number of cells entered G_0/G_1 phase in the Rg_1 group were increased, but the colony-formed ability was decreased, Rg_1 could significantly inhibit the proliferation and self-renewal ability of CD34~+CD38~-LSCs. The expression of SIRT1 and TSC_2 mRNA and protein were down regulated in the Rg_1 group compared with the control group. Our research implied that Rg_1 may induce the senescence of CD34~+CD38~-LSCs and SIRT1/TSC_2 signal axis plays a significant role in this process.
Cellular Senescence ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Leukemia, Myeloid, Acute ; Neoplastic Stem Cells ; drug effects ; Signal Transduction ; Sirtuin 1 ; metabolism ; Tuberous Sclerosis Complex 2 Protein ; metabolism ; Tumor Cells, Cultured

OBJECTIVE：To establish the method for determining protein binding rate of dipeptidyl peptidase- 4 inhibitor LGT- 6 in different species of plasma ，and to compare their difference. METHODS ：By equilibrium dialysis ，LGT-6（3，30，300，3 000 nmol/L）was equilibrated in rat ，monkey and human plasma （i. e. internal dialysis solution ）for 48 h，using phosphate buffer as the external dialysis solution. The concentration of LGT- 6 in internal and external dialysis solution was determined by UPLC-MS/MS using tolbutamide as internal standard ，and the plasma protein binding rate was calculated. The determination was performed on ACQUITY UPLC HSS T 3 column with water （containing 0.01％ formic acid ）-acetonitrile（containing 0.01％ formic acid ）as mobile phase at the flow rate of 0.6 mL/min. The column temperature was 40 ℃，and the sample size was 2 μL. The ion source was electrospray ion source ，and the multiple ion monitoring mode was used to carry out positive ionization scanning. The ion pairs for quantitative analysis were m/z 487.0→434.3（LGT-6），m/z 271.1→172.0（internal standard ），respectively. RESULTS ：At the concentrations of 3，30，300，and 3 000 nmol/L，the protein binding rates of LGT- 6 in rat plasma were （96.25±0.97）％，（84.16± 1.24）％，（78.25±0.61）％，（66.63±0.95）％；the protein protein binding rates in monkey plasma were （98.54±0.58）％，（87.27± 1.01）％，（79.35±0.86）％，（66.69±0.54）％；the protein binding rates in human plasma were （99.40±1.03）％，（84.48± 1.15）％，（77.62±0.77）％，（66.93±0.48）％. At the same concentration ，the protein binding rates of LGT- 6 in rat ，monkey and human plasma had no significant difference （P＞0.05）. In the same species of plasma ，there were significant differences in the plasma protein binding rates of different concentration of LGT-6 among those groups （P＜0.05），and it decreased with 才〔2016〕4015） the increase of drug concentration. CONCLUSIONS ： The method for the determination of plasma protein binding rate of LGT-6 is successfully established. The data revealed that the protein binding rate of LGT- 6 is concentration-dependent ， there was no obvious spec ies difference on protein binding rates of LGT- 6 in rat ，monkey and human plasma under the same concentration.

OBJECTIVE： To observe improvement effects of fingolimod on middle cerebral artery occlusion/reperfusion （MCAO/R） injury model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group and fingolimod low-dose， medium-dose and high-dose groups （0.5， 1， 2 mg/kg）， with 8 rats in each group. Except for sham operation group， MCAO/R injury model was induced by suture-occluded method in other groups. Administration groups were given relevant medicine intragastrically after reperfusion [1 h after reperfusion （1st day）， 22.5 h after reperfusion （2nd day）， and then every 24 h until 142.5 h of reperfusion （7th day）]. Sham operation group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 7 d. The scores of neurological deficit and balance beam test， the times of memory error [work memory error （WME）， reference memory error （RME） and total error] were recorded in each group. The contents of serum inflammatory cytokines （IL-6， IL-8， IL-10， TNF-α） were determined by ELISA， and triphenyl tetrazolium chloride staining method was used to detect the rate of cerebral infarction. RESULTS： Compared with sham operation group， neurological deficit scores （at different time points of 1st-7th day after administration）， balance beam test scores （2nd， 4th， 7th day after administration）， times of memory error （2nd， 4th， 7th day after administration）， the contents of serum inflammatory cytokines and the rate of cerebral infarction were increased significantly in model group （P＜0.05 or P＜0.01）. Compared with model group， neurological deficit scores （low-dose group at different time points of 3rd-7th day， medium-dose and high-dose groups at different time points of 2nd-7th day after administration）， balance beam test scores （low-dose group at 7th day， medium-dose group at 4th and 7th day， high-dose group at 2nd， 4th， 7th day）， RME times and total error times （low-dose group at 4th and 7th day， medium-dose group and high-dose group at 2nd， 4th， 7th day after administration）， WME times （administrations groups at 7th day after administration）， serum contents of IL-6， IL-8 and IL-10 （administrations groups）， serum contents of TNF-α （medium-dose and high-does groups） and cerebral infarction rate （medium-dose and high-dose groups） were all decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Intragastric administration of fingolimod can significantly reduce neurological deficit score， balance beam test score and the times of memory error in MCAO/R injury model rats， and has a protective effect on cerebral tissue and memory function. These effects may be related to the down-regulation of inflammatory cytokines such as IL-6 and TNF-α by fingolimod.

OBJECTIVE：To prepare Imperatorin ultradeformable liposomes gel （IMP-UDLs-Gel），and to evaluate its quality. METHODS：Based on single factor test ，using 12 h accumulative penetration amount （Q12h）as evaluation index ，the proportion of carbomer 940，glycerol and propyl glycol in formulation of IMP-UDLs-Gel were investigated by orthogonal test. The optimal formulation was screened. The quality of IMP-UDLs-Gel prepared with the optimal formulation was evaluated. RESULTS ：The optimal formulation of IMP-UDLs-Gel included carbomer 940 proportion of 1％，glycerol proportion of 15％ and propyl glycol proportion of 10％. Q12 h of IMP-UDLs-Gel was （11.543±0.241）μg/cm2；the appearance was milky white and translucent ；the particle size was （93.13±1.68）nm，PDI was 0.268±0.012，Zeta potential was （－24.96±1.99）mV；pH was 7.32±0.03； viscosity was （45.37±1.27）g·s；steady flow was （0.727±0.002）μg·h/cm2，lag time was （4.358±0.175）h，apparent permeability coefficient was 1.392×10－3 cm/h，and it has good physical and optical stability. CONCLUSIONS ：The preparation method is stable and feasible ，and the prepared IMP-UDLs-Gel has good adhesion ，stability and transdermal property.

OBJECTIVETo evaluate the safety of a group A + C meningococcal polysaccharide vaccine as part of a phase IV clinical trial.

METHODSThe study area was divided into 108 clusters according to the principle of cluster randomization, stratified and paired sampling methods. 54 out of 108 clusters served as observation groups were administered A + C vaccine, while the rest 54 groups were administered Vi polysaccharide vaccine. An adverse event surveillance system was established to monitor the adverse events following the vaccination campaign. Identical form and methods were used for data collection to investigate the adverse events following the vaccination of both A+ C vaccine and Vi vaccine.

RESULTS34,543 people were vaccinated, including 18,167 of whom received A + C vaccine, while the other 16,376 received Vi vaccine. The rates of immediate injection reaction and unsolicited non-serious adverse events from A + C vaccine group were 0.44% and 0.38% while of Vi vaccine group were 0.79% and 0.73% respectively. At the solicited adverse event survey on 3-day-post-vaccination, 1239 vaccinees were followed-up including 771 received A + C vaccine and 468 received Vi vaccine. The local injection reaction rate of A + C vaccine group on the 1st day was significantly higher (X2 = 13.98, P = 0.0002) than that of Vi vaccine group. Neither the local injection reaction rate nor the system reaction rate between both groups was significantly different on 2nd and 3rd day, post vaccination. It was not statistically different when comparing fever onset rate between those who received vaccine and those who did not, in each vaccine group. There were no serious adverse events observed.

CONCLUSIONResults showed that the side effects of A + C vaccine and the Vi vaccine were mild and safe for vaccination campaigns targeting on populations at different age.

Adolescent ; Adult ; Age Distribution ; Child ; Child, Preschool ; Female ; Humans ; Male ; Meningococcal Vaccines ; adverse effects ; immunology ; Middle Aged ; Polysaccharides, Bacterial ; immunology ; Sex Distribution ; Young Adult