Objective To develop a method of quantitative analysis of multi-components by single marker( QAMS) for determination of quercetin, kaempferide,isohamnetin in Chuipencao granules,providing method to control the quality of Chuipencao granules.Methods The analysis was performed at 30 ℃,on an Shimadzu C18 column(250 mm ×4.6 mm,5μm),and the mobile phase was methanol-0.4% phosphoric acid solution .The flow rate was 1.0 mL/min, and the injection volume was 10 μL.Quercetin as a reference to establish the relative correction factor kaempferol and between isorhmnetin, the method was evaluated for reproducibility, and the difference between calculated and measured values was compared. Results Quercetin, kaempferol and isorhmnetin respectively 32.36-426.43μg, 11.56-136.87μg, 10.45-155.68μg showed a good linear relationship, and the regression equation was: Y=3625.8X-13658 (R2 =0.9998), Y=2682.9X-10253(R2 =0.9999), Y=3012.2X-11223 (R2 =0.9999).The average recoveries were 99.3%, 99.6%, 98.5% (RSD were less than 1.0%).Determination of 10 batches of Chuipencao granules with QAMS and external standard method, kaempferol and isorhamnetin were measured, and measured values were basically the same.Conclusion The QAMS method is reliable and accurate, which might be used for the quality control of Chuipencao granules.

Objective To observe the influence of Kuailvning capsule(KLN)and its drug serum on the concentration of Ca2 + in rat ventricular muscle cells.Methods SD rats were randomly divided into five groups:the control group,serum control group,KLN high -dose group,KLN medium -dose group and KLN low -dose group,six mice in each group.The rat ventricular myocytes were separated by enzymatic hydrolysis.The influence of KLN and its drug serum on the concentration of Ca2 + in rat ventricular muscle cells were observed by Fluo -3 /AMprobe.Results The mean fluorescence intensity of cytosolic Ca2 + in the KLN high -dose group,KLN medium -dose group and KLN low -dose group after 50s,100s,150s,200s,250s were [(18.75 ±1.55),(16.69 ±0.93),(17.76 ±1.26)], [(25.47 ±1.118),(17.86 ±1.49),(17.81 ±1.13)],[(29.05 ±1.31),(20.14 ±1.73),(18.26 ±1.37)], [(35.21 ±1.33),(23.19 ±0.97),(18.18 ±1.46)],[(41.08 ±1.21),(26.34 ±1.69),(17.91 ±1.01)], which were higher than those in the control group(F =5.556,7.007,8.816,10.208,12.232,all P <0.01 ).The mean fluorescence intensity of cytosolic Ca2 + in the KLN high -dose group after 50s,100s,150s,200s,250s were (17.85 ±1.69),(18.95 ±1.73),(21.85 ±1.39),(23.01 ±1.48),(24.07 ±1.42),which were higher than those in the serum control group(t =3.642,4.406,6.074,7.402,8.625,all P <0.01 ).The mean fluorescence intensity of cytosolic Ca2 + in the KLN medium -dose group after 100s,150s,200s,250s were (18.01 ±1.42), (18.76 ±1.22),(19.73 ±1.37),(21.04 ±1.16),which were higher than those in the serum control group(t =3.902,4.033,4.126,4.326,all P <0.01).There were no statistically significant difference between the KLN low -dose group and the serum control group(P >0.05).Conclusion KLN drug serum can promote myocardial cell spon-taneous extracellular calcium influx,the mechanism of KLN antiarrhythmic effect may be related to the Ca2 + channel.

Objective To explore the MSCT manifestations of chromophobe renal cell carcinoma (CRCC)and to improve its accu-racy of preoperative diagnose.Methods Clinical and MSCT finding were retrospectively reviewed in 14 patients with CRCC,which were confirmed by surgical pathology or biopsy.Results (1)On plain scanning,most of the CRCC lesions showed round or oval (86.7%,13/1 5)and isodensity (73.3%,1 1/1 5)mass;and the border of 66.7% (10/1 5)lesions were clear.53.3% (8/1 5)of the lesions were heterogeneous with lamellar cystic low density (8 lesions)and coarse ringed and/or foliated calcifications (4 lesions). (2)On contrast-enhanced sacanning,the CRCC lesions showed mild to moderate (86.7%,13/1 5)and heterogeneous (60%,9/1 5) enhancement,and 73.3% (1 1/1 5)of the lesions were persistent enhanced.(3)According to the pathology,80% (12/1 5)of the le-sions was the typical type,20% (3/1 5)was the eosinophilic type,and 0 was the hybrid type.Conclusion CRCC demonstrates cer-tain characteristics signs at MSCT examination.Lesion mostly shows well-circumscribed round or oval mass in the renal parenchy-ma,mild to moderate,heterogeneous and continuous enhancement on the contrast scanning.The diagnosis of CRCC should be con-sidered especially when the lesion has cystic change and coarse ringed and/or foliated calcifications.

Objective To investigate the influence of ionic strength on the stability of the methotrexate-loaded dendrimer nanoparticles. Methods The influences of different ions (Na+,Cl-) and different concentrations of sodium chloride on the stability of the nanoparticles were studied. The particle size was measured by dynamic light scattering(DLS) and drug-loading content was determined by high-performance liquid chromatography (HPLC) in order to evaluate the stability. Results The Cl- was finally verified to play an important role in stabilizing the nanoparticles and the effective concentration of the sodium chloride was recommended to be below 1. 80% . Conclusion The recommended concentration (less than 1. 80% ) of the sodium chloride significantly improves the stability of the nanoparticles and benefits for long term storage.

Objective To enhance the solubility and bioavailability of curcumin (CUR).Methods A novel curcumin nanoparticles were prepared.The CUR-PGD nanoparticles were prepared by the method of ultrasound precipitation combined with high-pressure homogenization using codendrimer PAMAM-co-0.25OEG (PGD) as stabilizer.The stability of CUR-PGD nanoparticles was measured in 0.9% sodium chloride solution,5% glucose, PBS and plasma.Results The drug loading capacity (DL%) of CUR-PGD nanoparticles was 41.2%, the solubility of CUR was increased to 1.5 mg·mL-1 (50 times of CUR bulk powder).The mean diameter of the nanoparticles was 438.0 nm with spherical morphology and the zeta potential was 41.4 mV.The nanoparticles was stable in 0.9% sodium chloride solution,5% glucose, PBS and plasma and there was no hemolytic phenomenon, which meant they were suitable for intravenous administration.The DSC and XRD spectra of CUR-PGD nanoparticles showed that the CUR was presented as crystal morphology in the nanoparticles.The CUR released from nanoparticles was detected in different releasing medium and presented obvious controlled release behavior.Conclusion PGD may be an effective stabilizer for the preparation of CUR-PGD nanoparticles and CUR-PGD nanoparticles are a promising drug delivery system for CUR application in clinic.

Objective To observe the role and related mechanism of chemerin and its receptor ChemR23 in glomerular endothelial cells (GEnCs) stimulated by high glucose.Methods Mouse GEnCs were cultured and divided into control group,20.0 mmol/L high glucose group,40.0 mmol/L high glucose group and mannitol control group.Then the expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cell culture supematant as well as the expressions of intracellular protein and mRNA of chemerin,ChemR23,IL-6 and TNF-α were detected.Lentiviral transfection targeting ChemR23 was applied before high glucose-or Chemerin-stimulated,and expressions of supernatant and intracellular mRNA of IL-6 and TNF-α were measured.Meanwhile whether p38 mitogen-activated protein kinase (p38 MAPK) pathway was activated by high glucose was detected.The specific inhibitor of p38 MAPK was added prior to high glucose-stimulated,then supernatant and intracellular mRNA expressions of IL-6 and TNF-α was detected.The supernatant expressions of IL-6 and TNF-α were measured by ELISA.The intracellular protein expression and p38 MAPK phosphorylation activity were detected by Western blotting.The mRNA expression was detected by real time PCR.Results Compared with those in the control group,in high glucose groups the expressions of IL-6,TNF-α and chemerin were significantly increased (all P ＜ 0.05),however,the expressions of ChemR23 did not change (all P ＞ 0.05);the supernatant and mRNA expressions of IL-6 and TNF-α were also elevated in the chemerin group (all P ＜ 0.05).Lentivirus baring shRNA could efficiently suppress ChemR23 expression,and the Chemerin-or high glucose-induced expressions of IL-6 and TNF-α were reduced (all P ＜ 0.05).Also it could significantly reduce the expression of phosphorylated-p38 MAPK (p-p38 MAPK) induced by high glucose (P ＜ 0.05),as high glucose group had higher p-p38 MAPK than control group (P ＜ 0.05).While the high glucose-elevated expressions of IL-6 and TNF-α were significantly attenuated by p38 MAPK inhibitor (all P ＜ 0.05).Conclusions High glucose stimulation can induce the expression of chemerin in GEnCs.By binding to ChemR23,chemerin activates p38 MAPK signaling pathway,and then promotes the expressions of IL-6 and TNF-α.These inflammatory cytokines aggravate inflammation of GEnCs.

OBJECTIVE: To investigate the effects of gene of phosphate and tension homology (PTEN)-induced putative kinase 1 (PINK1)/Parkin pathway on hippocampal mitophagy and cognitive function in mice with sepsis-associated encephalopathy (SAE) and its possible mechanism. METHODS: A total of 80 male C57BL/6J mice were randomly divided into Sham group, cecal ligation puncture (CLP) group, PINK1 plasmid transfection pretreatment groups (p-PINK1+Sham group, p-PINK1+CLP group), empty vector plasmid transfection control group (p-vector+CLP group), with 16 mice in each group. The mice in CLP groups were treated with CLP to reproduce SAE models. The mice in the Sham groups were performed laparotomy only. Animals in the p-PINK1+Sham and p-PINK1+CLP groups were transfected with PINK1 plasmid through the lateral ventricle at 24 hours before surgery, while mice in the p-vector+CLP group were transfected with the empty plasmid. Morris water maze experiment was performed 7 days after CLP. The hippocampal tissues were collected, the pathological changes were observed under a light microscope after hematoxylin-eosin (HE) staining, and the mitochondrial autophagy was observed under a transmission electron microscopy after uranyl acetate and lead citrate staining. The expressions of PINK1, Parkin, Beclin1, interleukins (IL-6, IL-1β) and microtubule-associated protein 1 light chain 3 (LC3) were detected by Western blotting. RESULTS: Compared with the Sham group, CLP group mice in Morris water maze experiment had longer escape latency, shorter target quadrant residence time, and fewer times of crossing the platform at 1-4 days. Under the light microscope, the hippocampal structure of the mouse was injured, the neuronal cells were arranged in disorder, and the nuclei were pyknotic. Under the electron microscope, the mitochondria appeared swollen, round, and wrapped by bilayer or multilayer membrane structures. Compared with the Sham group, CLP group had higher expressions of PINK1, Parkin, Beclin1, LC3II/LC3I ratio, IL-6 and IL-1β in hippocampus, indicating that sepsis induced by CLP could activated inflammatory response and caused PINK1/Parkin-mediated mitophagy. Compared with the CLP group, p-PINK1+CLP group had shorter escape latencies, spent more time in the target quadrant and had more number of crossings in the target quadrant at 1-4 days. Under the light microscope, the hippocampal structures of mice was destroyed, the neurons were arranged disorderly, and the nuclei were pyknotic. Under transmission electron microscope, swollen and rounded mitochondria and mitochondrial structure wrapped by double membrane or multilayer membrane structure were observed. Compared with the CLP group, the levels of PINK1, Parkin, Beclin1 and LC3II/LC3 ratio in the p-PINK1+CLP group were significantly increased [PINK1 protein (PINK1/β-actin): 1.95±0.17 vs. 1.74±0.15, Parkin protein (Parkin/β-actin): 2.06±0.11 vs. 1.78±0.12, Beclin1 protein (Beclin1/β-actin): 2.11±0.12 vs. 1.67±0.10, LC3II/LC3I ratio: 3.63±0.12 vs. 2.27±0.10, all P < 0.05], while the levels of IL-6 and IL-1β were significantly decreased [IL-6 protein (IL-6/β-actin): 1.69±0.09 vs. 2.00±0.11, IL-1β protein (IL-1β/β-actin): 1.11±0.12 vs. 1.65±0.12, both P < 0.05], suggesting that overexpression of PINK1 protein could further activate mitophagy and reduce the inflammatory response caused by sepsis. There was no statistically significant difference in the above pathological changes and related indicators between Sham group and p-PINK1+Sham group, CLP group and p-vector+CLP group. CONCLUSIONS PINK1 overexpression can further activate CLP-induced mitophagy by upregulating Parkin, thereby inhibiting inflammation response and alleviate cognitive function impairment in SAE mice.
Male ; Animals ; Mice ; Mice, Inbred C57BL ; Sepsis-Associated Encephalopathy ; Phosphates ; Actins ; Beclin-1 ; Interleukin-6 ; Autophagy ; Ubiquitin-Protein Ligases ; Cognitive Dysfunction ; Sepsis ; Mitochondria ; Protein Kinases

Purpose To evaluate left ventricular systolic function of patients with obstructive sleep apnea syndrome (OSAS).Materials and Methods Sixty OSAS patients were divided into 16 cases in mild OSAS group,18 cases in medium OSAS group and 26 cases in severe OSAS group in line with polysomnography results.20 healthy people were in the control group.Two-dimensional speckle tracking imaging was adopted to measure left ventricular global longitudinal strain (GLS),global radial strain (GRS),global circumferential strain (GCS),Endo-LS,Mid-LS and Epi-LS of left ventricular long axial section and EndoCS,Mid-CS and Epi-CS of short axial section in OSAS group and control group.Results ① There was no obvious change of GLS and GCS in mild OSAS group and compared with those in control group (P＞0.05),GRS was increased (P＜0.05).GLS,GRS and GCS in medium OSAS group was decreased when compared with those in control group,and the difference was of statistical significance (P＜0.05).GLS in severe OSAS group was reduced compared with that in control group,mild and medium OSAS group and GRS and GCS were also reduced compared with that in control group and mild OSAS group.The difference was of statistical significance (P＜0.05).② OSAS group was represented by Endo-LS reduction and as OSAS worsened,Endo-LS reduced section was enhanced.③ Starting from medium OSAS group,myocardial circumference strain began to decrease,which was represented by Endo-CS and Mid-CS reduction.Conclusion Adoption of two-dimensional speckle tracking imaging for left ventricular global and layered myocardial strain could figure out early stage left ventricular systolic function disorder in OSAS patients,providing more valuable information for clinical activities and guidance of early intervention for OSAS patients.

Objective To imvestigate the application value of combined newborn deafness genetic and hearing screening in the prevention of deafness in poor families in Ningxia.Methods Heel blood was collected from 3 023 neonates in Guyuan City,and 15 loci of 4 deafness genes were detected by gene chip technology of hereditary deaf-ness.At the meantime,heaning was screened and followed-up in all newborns.Results Among the 3 023 neo-nates,123 were positive for deafness gene screening(4.07％,123/3 023),including 56 cases with GJB2 mutation(1.85％,56/3 023),46 cases with SLC26A4 mutation(1.52％,46/3 023),6 cases with GJB3 mutation(0.20％,6/3 023),14 cases with mtDNA12SrRNA(0.46％,14/3 023).The mutation detection rates of c.235delC and c.IVS7-2A＞G loci were 1.36％(41/3 023)and 0.93％(28/3 023)respectively,which were the main mutation types.A total of 98 cases were found in Hui nationality to carry deafness gene mutation,with a carrying rate of 4.26％(98/2 302).A total of 25 mutations were detected in the Han nationality(3.49％,25/715).The total mu-tation rate of four common deafness mutation genes between Hui and Han was not significantly different(P＞0.05).All 123 newborns with deafness gene mutation passed the hearing screening(100％).The hearing screening passing rate of 690 Han newborns with negative deafness gene screening results was 99.71％(688/690),and the hearing screening passing tate of 2 204 Hui newborns with negative deafness gene screening results was 99.86％(2 201/2 204).There was no significant difference in the failure rate of hearing screening between Hui and Han newborns with positive deafness gene screening(P＞0.05).All 3 023 neonates completed follow-up(100％).Five neonates failed to pass the hearing re-examination,and 3 neonates were diagnosed with hearing loss.The hearing follow-up of 123 neonates with positive deafness gene mutation showed normal hearing and language development.Conclusion GJB2:c.235delC and SLC26A4:c.IVS7-2A＞G are the main pathogenic gene mutations in the neonates of poor and registered households in Guyuan area.The mitochondrial 12SrRNA carrying rate in Han neonates is higher than that in Hui neonates.

Objective To investigate the changes in Th17 cells and CD4+CD25+regulatory T lym-phocytes ( Treg) as well as transcription factors and cytokines relating to them in children with Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (HLH) and to analyze their role and clinical significance. Methods Thirty-two children with newly diagnosed EBV-associated HLH in the Hematology/Oncology Department of Zhengzhou Children′s Hospital from January 2012 to December 2016 were enrolled in this study. Thirty healthy children taking physical examination in the same hospital in the corresponding period were recruited as controls. Percentages of Th17 and Treg cells in peripheral blood T lymphocytes were detected by flow cytometry. Expression of RORγt and Foxp3 at mRNA level in peripheral blood mononuclear cells was detected by real-time PCR. Levels of IL-6, IL-17, IL-10 and TGF-β1 in serum samples were measured by ELISA. Results Compared with the control group, the EBV-associated HLH group showed in-creased percentage of Th17 cells [(1. 09±0. 43)％ vs (0. 39±0. 19)％, P<0. 05] and enhanced expres-sion of RORγt at mRNA level [(1. 41±0. 37) vs (0. 67±0. 13), P<0. 05], but decreased percentage of Treg cells [(3. 66±1. 13)％ vs (6. 80±1. 15)％, P<0. 05] and inhibited expression of Foxp3 at mRNA level [(15. 97±5. 11) vs (30. 23±4. 95), P<0. 05]. All of the above mentioned changes were reversed af-ter treatment (P<0. 05). Serum levels of IL-6 and IL-17 of EBV-associated HLH group were higher than those of control group, while serum levels of IL-10 and TGF-β1 were lower (P<0. 05). Conclusion Im-balanced Th17/Treg cells might play an important role in the pathogenesis of EBV-associated HLH. Cyto-kines relating to the maintenance of Th17/Treg cell balance could be used as indicators of disease develop-ment.