Adult ; Aged ; Brain ; diagnostic imaging ; physiopathology ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Optic Neuropathy, Ischemic ; diagnostic imaging ; physiopathology

To investigate the differences of chemical compositions in Gynostemma pentaphyllum leaves prepared by different processing methods. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to compare the chemical compositions between shade-dried processing and drum-dried processing. Forty six gypenosides were identified by control comparison， liquid chromatography-mass spectrometry(LC-MSn) fragmentation information， and literature data. The mass spectral peak area statistics was combined with principal component analysis(PCA)， and the results showed that eight batches of Gynostemma pentaphyllum leaves samples were divided into two groups according to the two different processing methods; ten chemical compositions with significant differences were screened according to mass spectrum information combined with partial least-squares discriminant analysis(PLS-DA). The result showed that most parent nucleus of the gypenosides contained three to four glycosides in drum-dried samples， and one to two glycosides in the shade-dried samples. It was inferred from further MS analysis that desugarization of gypenosides was present to produce secondary glycosides with the effect of glucosidase in the shade-drying， thus resulting in difference in compositions. This study provided data support for harvesting， processing and quality control of Gynostemma pentaphyllum leaves.
Chromatography, High Pressure Liquid ; Gynostemma ; chemistry ; Mass Spectrometry ; Plant Leaves ; chemistry ; Saponins ; chemistry ; isolation & purification

As an outstanding representative of traditional Chinese medicine(TCM) prescriptions accumulated from famous TCM doctors' clinical experiences in past dynasties, classical TCM excellent prescriptions (cTCMeP) are the most valuable part of TCM system. To support the research and development of cTCMeP, a series of regulations and measures were issued to encourage its simplified registration. There is still a long-way to go because many key problems and puzzles about technology, registration and administration in cTCMeP R&D process are not resolved. Based on the analysis of registration and management regulations of botanical drug products in FDA of USA and Japan, and EMA of Europe, the possible key problems and countermeasures in chemistry, manufacture and control (CMC) of simplified registration of cTCMeP were analyzed on the consideration of its actual situation. The method of "reference decoction extract by traditional prescription" (RDETP) was firstly proposed as standard to evaluate the quality and preparation uniformity between the new developing product under simplified registration and traditional original usages of cTCMeP, instead of Standard Decoction method in Japan. "Totality of the evidence" approach, mass balance and bioassay/biological assay of cTCMeP were emphatically suggested to introduce to the quality uniformity evaluation system in the raw drug material, drug substance and final product between the modern product and traditional decoction.

OBJECTIVETo investigate the prenatal diagnosis of a case of SEA-HPFH deletion combined with beta-thalassemia in a Chinese family.

METHODSGap-PCR and RDB methods were applied to test the genotype for the family.

RESULTSMother showed a SEA-HPFH thalasemia trait phenotype, while her genotype was heterozygote for SEA-HPFH deletion; father showed a beta-thalassemia trait phenotype, while his genotype was heterozygote for IVS-II-654 mutation; the genotype of fetus was normal in these tests.

CONCLUSIONRegular thalassemia genes and deletion beta-thalassemia genes can be used in prenatal diagnosis of the case at risk for compound heterozygotes of SEA-HPFH deletion and beta-thalassemia.

To optimize the purification process of gynostemma pentaphyllum saponins (GPS) based on "adjoint marker" online control technology with GPS as the testing index. UPLC-QTOF-MS technology was used for qualitative analysis. "Adjoint marker" online control results showed that the end point of load sample was that the UV absorbance of effluent liquid was equal to half of that of load sample solution, and the absorbance was basically stable when the end point was stable. In UPLC-QTOF-MS qualitative analysis, 16 saponins were identified from GPS, including 13 known gynostemma saponins and 3 new saponins. This optimized method was proved to be simple, scientific, reasonable, easy for online determination, real-time record, and can be better applied to the mass production and automation of production. The results of qualitative analysis indicated that the "adjoint marker" online control technology can well retain main efficacy components of medicinal materials, and provide analysis tools for the process control and quality traceability.

Objective To explore the biological functions of E77.43, a gene segment of Microtus fortis, in treating Schistoso-ma japonicum infection. Methods Recombinant retroviral vectors of pRevTRE-E77.43 was constructed, and recombinant retro-viral vectors were transfected into PA317 cells, and the stable cell lines were obtained by hygromycin screening, followed by the packaging, concentration and purification of recombinant retrovirus. The virus was transferred to the mice infected by S. japoni-cum via intravenous or intraperitoneal injection, through which the express of target gene and the treatment function in vivo were observed. Results The experiment showed the recombinant virus injected mice could efficiently express E77.43 on the 7th day after the injection which lasted for forty-five days thereafter. A significant reduction in adult worms (31.0％) and a high reduction (35.0％) in liver eggs were induced by pRevTRE-E77.43, while the reduction in adult worms and that in liver eggs was 1.2％and 0.9％induced by pRevTRE respectively (t=3.524, 9.485, both P<0.01). Conclusion pRevTRE-E77.43 could be used for the treatment of S. japonicum infection, indicating that E77.43 may involve in the natural resistance of M. fortis to S. japonicum infec-tion.

Property and flavor theory of traditional Chinese medicine (TCM) is the core base for clinical treatment of diseases. However, few research about its chemical and biological characterization was performed. In this paper, network pharmacology was adopted to review patterns around the theory of TCM. "Xiaoke" prescription database, which combinations of herb medicines for diabetes therapy, was firstly built to explore prescription regularity and screen core paired-components. The prescription regularity and molecular mechanism of flavor composition were explored through the relationship of "drug-compound-target-pathway-function" by ChEMBL, CTD and KEGG datebase. As a result, the tastes of "Gan" (sweetish taste) and "Ku" (bitter taste) were the popular therapeutic flavor to regulate the disorder of glucose and lipid metabolisms. The mechanism of Xiaoke was summarized from representative traditional Chinese medicine partner "Zhimu-Huangbai" and "Huangqi-Gegen". The key components of "Gan", including saponins stimulated insulin secretion, improve insulin resistance and promote glucose utilization. The components of "Ku", including flavonoids and alkaloids regulate inflammatory cytokines, promoted the utilization of glucose, improve endocrine and metabolism through MAPK, PI3K-Akt, PPAR signal pathway. The TCM therapeutic mechanism about "Xiaoke" was preliminarily summarized to clear "heat" by anti-inflammation and immunoregulation, to regulate glucolipid metabolism for removing the satiation of digestion, and to improve the utilization of insulin and diabetes complications for endocrine adjusting. The results demonstrate that therapeutic principle of TCM for "Xiaoke" is comprehensive via multi pathway. This study provides a new research method and strategy for exploring the mechanism of TCM for diabetes therapy.

BACKGROUNDUrine output (UO) is an essential criterion of the Kidney Disease Improving Global Outcomes (KDIGO) definition and classification system for acute kidney injury (AKI), of which the diagnostic value has not been extensively studied. We aimed to determine whether AKI based on KDIGO UO criteria (KDIGOUO) could improve the diagnostic and prognostic accuracy, compared with KDIGO serum creatinine criteria (KDIGOSCr).

METHODSWe conducted a secondary analysis of the database of a previous study conducted by China Critical Care Clinical Trial Group (CCCCTG), which was a 2-month prospective cohort study (July 1, 2009 to August 31, 2009) involving 3063 patients in 22 tertiary Intensive Care Units in Mainland of China. AKI was diagnosed and classified separately based on KDIGOUOand KDIGOSCr. Hospital mortality of patients with more severe AKI classification based on KDIGOUOwas compared with other patients by univariate and multivariate regression analyses.

RESULTSThe prevalence of AKI increased from 52.4% based on KDIGOSCrto 55.4% based on KDIGOSCrcombined with KDIGOUO. KDIGOUOalso resulted in an upgrade of AKI classification in 7.3% of patients, representing those with more severe AKI classification based on KDIGOUO. Compared with non-AKI patients or those with maximum AKI classification by KDIGOSCr, those with maximum AKI classification by KDIGOUOhad a significantly higher hospital mortality of 58.4% (odds ratio [OR]: 7.580, 95% confidence interval [CI]: 4.141-13.873, P< 0.001). In a multivariate logistic regression analysis, AKI based on KDIGOUO (OR: 2.891, 95% CI: 1.964-4.254, P< 0.001), but not based on KDIGOSCr (OR: 1.322, 95% CI: 0.902-1.939, P = 0.152), was an independent risk factor for hospital mortality.

CONCLUSIONUO was a criterion with additional value beyond creatinine criterion for AKI diagnosis and classification, which can help identify a group of patients with high risk of death.

Acute Disease ; mortality ; Aged ; Creatinine ; blood ; Critical Illness ; mortality ; Female ; Hospital Mortality ; Humans ; Kaplan-Meier Estimate ; Kidney Diseases ; blood ; mortality ; pathology ; urine ; Logistic Models ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Risk Factors

Objective To study the basic regular patterns of stress distributions inside and outside periacetabular districts during normal gait cycle of healthy adults, so as to provide clinical guidance for acetabular reconstruction of total hip arthroplasty (THA). Methods Based on CT scans of a male and a female healthy adult volunteer, The three-dimensional model including pelvis and proximal femur was reconstructed. By using an inhomogeneous material distribution scheme which was based on CT data to calculate elastic modulus and convergence analysis, each element was given a corresponding material attribute. The dynamic change of hip contact force during a normal gait cycle was used as the load condition to the model. Von Mises stress of the nodes inside and outside the model was considered as the criterion to assess the results. Results During normal gait, the stress on the hip surface of two volunteers was mainly transmitted from postersuperior part of acetabulum to auricular surface along posterolateral of iliac wing, and the maximum stress was at the district near greater sciatic. As for the superior, middle and inferior section of two volunteers' acetabulum, the stress was distributed both on cortical and cancellous bone of postersuperior part. However, in terms of acetabular anterior and posterior column, the stress distribution was mainly found on cortical bone. Conclusions According to the observed acetabular stress distribution pattern of health adults during normal gait cycle, choosing acetabular component with more suitable size and controlling the placement of acetabular component with more accuracy could obtain some acetabular reconstruction plan better in accordance with stress distributions during normal gait.

OBJECTIVETo study the potential role of mast cells and the related molecular mechanism in chronic hepatitis (CH) using a rat model system.

METHODSThirty Wistar rats (15 males, 15 females; weight range: 230-290 g) were randomly divided into the normal contrast (NC) group and experimental CH group. The CH group received subcutaneous injection of CCl4 and a diet high in cholesterol and alcohol content and low in protein and choline content. Throughout the 4-week modeling period, aseptic blood samples were taken to test plasma tryptase (TS) and hyaluronic acid (HA) levels. The rats were euthanized to assess the changes in liver mast cells by histology and morphology analyses and the changes in liver expression of c-kit and stem cell factor (SCF) proteins by immunohistochemistry and mRNAs by RT-PCR.

RESULTSCompared to the NC group, the CH group had higher plasma and liver concentration of HA (78.09 +/- 38.55 vs. 145.14 +/- 52.54 ng/ml, 51.58 +/- 20.45 vs. 106.59 +/- 43.15 ng/100 mg; t = 2.457 and 2.825 respectively, both P less than 0.05) and TS (0.416 +/- 0.143 vs 0.753 +/- 0.210 mg/ml; t = 4.165, P less than 0.05). The CH group also showed fatty degeneration and fibrosis with many degranulating and degranulated mast cells filled with purple granula located around the liver blood vessels and in fiber-intervals. The CH livers also showed a significantly higher number of mast cells (2.167 +/- 0.924 vs. NC: 10.92 +/- 1.575; t = 7.633, P less than 0.05) and stronger intensity of c-kit staining (2.783 +/- 0.577 vs. 12.86 +/- 3.126; t = 9.511, P less than 0.05) and SCF staining (3.383 +/- 1.583 vs. 15.58 +/- 6.431; t = 9.625, P less than 0.05). The expressions of c-kit and SCF were positively correlated with HA level (r = 0.478 and 0.556 respectively, both P less than 0.05). The c-kit and SCF mRNA expression levels were also significantly higher in the CH liver tissues.

CONCLUSIONMast cell degranulation and histamine release is significantly increased under conditions of chronic hepatitis, and the related mechanism may involve up-regulation of the membrane receptor c-kit and its ligand SCF.

Animals ; Cell Degranulation ; Disease Models, Animal ; Female ; Hepatitis, Chronic ; metabolism ; pathology ; Hepatocytes ; metabolism ; Liver ; metabolism ; Liver Cirrhosis ; metabolism ; pathology ; Male ; Mast Cells ; metabolism ; physiology ; Proto-Oncogene Proteins c-kit ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Stem Cell Factor ; metabolism