OBJECTIVE To predict potential quality markers(Q-markers) in Yinhua Miyanling tablets based on fingerprinting and network pharmacology methods. METHODS HPLC fingerprints of 13 batches of Yinhua Miyanling tablets were established, and the similarity analysis was carried out using the "Chromatographic Fingerprint Evaluation System for Traditional Chinese Medicine" to identify the common peaks and attribute them. The fingerprints of Yinhua Miyanling tablets were investigated using chemometrics, cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis in combination with SPSS 26.0 and SIMCA 14.1 software to identify the major signature components responsible for the differences. The network pharmacology was used to screen and analyze the targets and pathways of Yinhua Miyanling tablets, construct a "drug-component-target-pathway" network diagram, and predict the Q-Marker and core targets of Yinhua Miyanling tablets. RESULTS HPLC fingerprint of Yinhua Miyanling tablets was established, and 27 common peaks including chlorogenic acid, mangostin, wild baicalin, lignocerin and quercetin were identified. Chemical pattern recognition analysis screened five components as differential markers for Yinhua Miyanling tablets. Five active ingredients, 20 core targets and 20 key pathways were screened by network pharmacology, showing that all five active ingredients could be used as potential Q-Markers. CONCLUSION The method is stable, accurate and feasible for screening five chemical components as potential Q-Markers for Yinhua Miyanling tablets. It provides a reference for the overall control of the quality of Yinhua Miyanling tablets, and also lays the foundation for further research on the mechanism of action of Yinhua Miyanling tablets.

OBJECTIVE： To rapidly identify chemical components of Zhitong huazheng capsules （shorted for “ZTHZC”）， and to provide reference for further elucidating the pharmacodynamic material basis and overall quality control. METHODS： UPLC-Q-TOF/MS method was adopted to separate the chemical components of ZTHZC. By reviewing literatures that related to chemical components of ZTHZC and its single Chinese traditional medicinal crops， the chemical structure of the chemical components was saved into a .mol format file， and supplemented the UNIFI database. The chemical constituents of ZTHZC were qualitatively analyzed by UNIFI database screening， and validated according to their precise molecular mass， characteristic ions， neutral loss， secondary mass spectrometry cleavage， chromatographic retention behavior， combined reference information and literature reports. RESULTS： A total of 70 chemical components were identified in this experiment， including 38 organic acids and organic acid esters， 8 alkaloids， 9 flavonoids， 5 triterpenoid saponins， 5 aldehydes， 2 ketones， 1 quinone， 1 polyacetylene and 1 monoterpenoid. CONCLUSIONS： This study provides a rapid detection and identification method for chemical components of ZTHZC， and also provides a scientific basis for the clarification of ZTHZC’s quality control and pharmacodynamics.

Objective SIV30 protein of simian immunodeficiency virus(SIV)was prepared by genetic engineering technique as an antigen diagnostic reagent, to establish an immune comb method for the specific detection of anti SIV IgG in monkey serum. Methods Recombinant expression plasmid of SIV SIV30 gene was constructed by prokaryotic expression vector pGEX-4T-1, and expressed in the competent BL21 cells. The recombinant protein was purified as a diagnostic antigen, and a standardized procedure for the detection of immune comb was established and applied for clinical detection. Results The optimum coating amount of antigen was 0.02 mg/mL. The prepared IC was able to specifically detect the positive serum of SIV. There was no cross reaction between the sera of other viruses. It showed a high specificity of the detection method. Sensitivity analysis showed that the SIV30 protein was able to detect 1:400 times diluted SIV positive sera. The result of stability and repeatability test(the same sample was repeated 3 times) showed that the coefficient of variation(CV)was less than 10%. The serum samples of 10 suspicious monkeys were detected by this method, showing a consistent rate of comb method and ELISA test result of 100%, Kappa =1.000. Conclusions SIV30 protein is expressed in prokaryotic cells. The immune comb is prepared,and is successfullyl applied in clinical examination. It shows that the method has a high sensitivity, strong specificity, good reproducibility and practicability.

Objective To explore the clinical application value of chemiluminescence detection of urine asymmetric dimethylarginine ( ADMA ) in pregnancy-induced hypertension ( PIH ) . Methods Collected the 24 h urine from 60 normal pregnancy women and 72 PIH pregnancy women who were admitted to Huzhou Maternal and Child Health Hospital from May 2014 to April 2015 by the case-control study , Determination of urine ADMA content by chemiluminescence ( CLIA ) and high performance liquid chromatography ( HPLC ) , the results between two assays were analyzed by the Rank sum test , receiver operating characteristic ( ROC) curve and pearson correlation analysis .Results Compared with the normal control group , the urine ADMA concentration in the PIH group was significantly increased by HPLC and CLIA, and the concentration of ADMA by CLIA in the PIH group was 68.18(57.25,81.55)μmol/L higher than that of the normal control group 30.11(22.69,42.97)μmol/L(Z=-8.139,P<0.001),and the concentration of ADMA by HPLC in the PIH group by HPLC was 71.11(57.65,82.89)μmol/L higher than that of the normal control group 28.11(21.06,42.99)μmol/L(Z=-8.356,P<0.001).The difference was statistically significant .The two methods of urine ADMA concentration were highly positively correlated with PIH blood pressure.The correlation coefficient r values were 0.746 and 0.763, respectively, the P values were 0.007 and 0.008 respectively.Conclusions CLIA can better detect the ADMA concentration in urine of pregnant women with PIH , and has a good clinical diagnosis ability .The ADMA concentration in urine is related to the blood pressure level of PIH .

OBJECTIVETo screen for genomic copy number variants (CNVs) in a fetus with cardiac abnormalities and intrauterine growth retardation through single nucleotide polymorphism microarray (SNP array) and karyotyping analysis.

METHODSThe fetus and its parents were subjected to conventional G banding and SNP-array analysis. The results were confirmed with fluorescence in situ hybridization (FISH).

RESULTSG-banding analysis showed that the fetus has a karyotype of 47,XX,+mar. The father has a karyotype of 46,XY,t(4;18) (p15.2q11.2), while the mother showed a normal karyotype. SNP-array detected two microduplications at 18p11.32q11.2 (20.5 Mb) and 4p16.3p15.2 (24.7 Mb) in the fetus. The supernumerary marker chromosome carried by the fetus has derived from the balanced translocation carried by its father. The result was confirmed by FISH.

CONCLUSIONBased on the two microduplications, the fetus was diagnosed as Wolf-Hirschhorn syndrome in conjunction with Edward syndrome. Verification of the origin of the supernumerary marker chromosome by SNP-array has provided a basis for prenatal genetic diagnosis.

Chromosome Banding ; Female ; Genetic Testing ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis ; Trisomy 18 Syndrome ; genetics ; Wolf-Hirschhorn Syndrome ; genetics

OBJECTIVETo screen for genomic copy number variants (CNVs) in a fetus with one sibling affected with Prader-Willi syndrome using single nucleotide polymorphism (SNP) array.

METHODSThe fetus and its parents were subjected to chromosomal karyotyping and SNP array analysis.

RESULTSA 5p15.33 microdeletions was identified in the fetus and its phenotypically normal mother with a size of 344 kb (113 576 to 457 213). The father was normal for both testing. Analysis of literature and CNVs database indicated the above CNV to be variant of unclear significance. The couple decided to continue with the pregnancy and gave birth to a healthy boy at full-term. No abnormalities were found during the follow-up.

CONCLUSIONThis study may provide further data for the phenotype-genotype correlation of 5p15.33 microdeletion, which differs from Cri du Chat syndrome.

Adult ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; genetics ; DNA Copy Number Variations ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Male ; Prader-Willi Syndrome ; diagnosis ; embryology ; genetics ; Pregnancy ; Prenatal Diagnosis

PURPOSE: Uridine-cytidine kinase (UCK) 2 is a rate-limiting enzyme involved in the salvage pathway of pyrimidine-nucleotide biosynthesis. Recent studies have shown that UCK2 is overexpressed in many types of cancer and may play a crucial role in activating antitumor prodrugs in human cancer cells. In the current study, we evaluated the potential prognostic value of UCK2 in breast cancer. METHODS: We searched public databases to explore associations between UCK2 gene expression and clinical parameters in patients with breast cancer. Gene set enrichment analysis (GSEA) was performed to identify biological pathways associated with UCK2 gene expression levels. Survival analyses were performed using 10 independent large-scale breast cancer microarray datasets. RESULTS: We found that UCK2 mRNA expression was elevated in breast cancer tissue compared with adjacent nontumorous tissue or breast tissue from healthy controls. High UCK2 levels were correlated with estrogen receptor negativity (p<0.001), advanced tumor grade (p<0.001), and poor tumor differentiation (p<0.001). GSEA revealed that UCK2-high breast cancers were enriched for gene sets associated with metastasis, progenitor-like phenotypes, and poor prognosis. Multivariable Cox proportional hazards regression analyses of microarray datasets verified that high UCK2 gene expression was associated with poor overall survival in a dose-response manner. The prognostic power of UCK2 was superior to that of TNM staging and comparable to that of multiple gene signatures. CONCLUSION: These findings suggest that UCK2 may be a promising prognostic biomarker for patients with breast cancer.
Biomarkers ; Breast Neoplasms* ; Breast* ; Dataset ; Estrogens ; Gene Expression ; Humans ; Neoplasm Metastasis ; Neoplasm Staging ; Phenotype ; Prodrugs ; Prognosis* ; RNA, Messenger ; Uridine Kinase*

Objective To explore the value of prenatal MRI in the diagnosis of fetal simple expansion of lateral ventricle(ventriculomegaly), and follow up the nervous system development status after birth. Methods Simple expansion of the lateral ventricle fetus by prenatal MRI examination were collected in Huzhou Maternal and Child Care Hospital from May 2013 to June 2015, 126 cases of live births in expansion group, 50 normal cases were recruited in the same period as the control group. In expansion group, fetal subgroup analysis was done:(1) unilateral or bilateral lateral ventricle expasion:one group was 98 cases was lateral ventricle expansion (77.8%, 98/126), expansion of bilateral ventricle group was 28 cases (22.2%, 28/126). (2) Prenatal MRI in the diagnosis of the lateral ventricle of expansion: expansion of the lateral ventricle width was greater than 10.0 mm, if both sides were expanding, the expand width was the heavier one side, divided into 3 subgroups: ①Expansion in group A (lateral ventricle width 10.0-12.0 mm) were 88 cases (69.8%, 88/126).②Expansion in group B (lateral ventricle width 12.1-15.0 mm) were 29 cases (23.0%, 29/126). ③Expansion of group C (lateral ventricle width> 15.0 mm) were 9 cases (7.12%, 9/126). All 176 cases were followed up after birth at the 3rd, 6th, 12th, 18th month (corrected age was used for premature babies), and Gesell developmental schedules (GDS) were used to evaluate the neurobehavioral development. Results (1) The MRI results after birth:21 cases were followed up by MRI after birth. In group A, 11 cases had MRI and 9 were normal (the ventricular width<10.0 mm after birth) , the other 2 cases were stable (the ventricular width measured first time after birth was ≥10.0 mm, but the difference was within 2.0 mm from the MRI before birth). In group B, 4 cases had MRI, 1 was normal, 1 was stable, and 2 cases were getting better (the ventricular width measured first time after birth was ≥10.0 mm, but the width decreased more than 2.0 mm from the MRI before birth). In group C, 6 cases had MRI. 3 cases were getting better and 3 cases were stable. (2) Overall GDS results:expansion group after the birth of the 3rd, 6th, 12th, 18th month GDS evaluation results compared with control group, respectively, the differences were not statistically significant (all P>0.05). (3) The GDS results among the subgroups:in each evaluation after birth, there were no statistically significant differences between group A and the control group (all P>0.05). The GDS results of group B at the 3rd and 6th month were lower than those of the control group (P<0.05); while there were no statistically significant differences between the 2 goups at the 12th and 18th month (P>0.05). And for group C, statistically significant differences were found compared to the control group at each follow-up time (all P<0.05). (4) GDS results at different times after birth in the expansion group:there was no statistically significant difference between the results at the 3rd and 6th month (P>0.05). But when the result at the 3rd month was compared to the results of the 12th or 18th month, the differences were statistically significant (P<0.05). GDS result of 6th months after birth compared with 12th and 18th months, respectively, there were no statistically significant differences (P>0.05). There was no statistically significant difference between the results at the 12th and 18th month (P>0.05). (5) The GDS results in unilateral and bilateral ventricle expansion:at the 18th month, among the 98 unilateral cases, 86 (87.8%, 86/98) had normal GDS results(>85 scores);8 (8.2%, 8/98) had borderline results (75-85 scores);4 (4.1%, 4/98) had delayed results (<75 scores). Among the 28 bilateral cases, 23 (82.1%, 23/28) had normal GDS results;3 (10.7%, 3/28) had borderline results; 2 (7.1%, 2/28) had delayed results. There was no statistically significant difference (P>0.05). Conclusions Among the simple expansion of lateral ventricle, those whose ventricular width are≤12.0 mm may not need clinical treatment. If the width is between 12.1 to 15.0 mm, closely follow-up and targeted rehabilitation training after birth are recommended. When the width is more than 15.0 mm, the risk of the central nervous system function delay is significantly increased, and early intervention might improve the prognosis.

Objective To evaluate the uncertainty of the pseudo-ginsenoside GQ (PGQ) concentration in human plasma by HPLC-MS/MS.Methods The whole process of PGQ determination by HPLC-MS/MS in human plasma was evaluated and the uncertainty caused by repeatability,weighing,standard solution preparation,biological sample preparation,extraction recovery process,recovery,instrument precision and calibration curve fitting were evaluated,respectively.The combined and expanded uncertainty values were both calculated.Results The expanded uncertainty values for low (15.16 ng·mL-1),medium (2 516.67 ng·mL-1) and high (3 902.00 ng·mL-1) levels of PGQ were 1.39,177.74 and 262.69 ng·mL-1,respectively (P =95 ％,k =2).Conclusion The uncertainty of the PGQ determination in human plasma by HPLC-MS/MS is mainly caused by recovery,repeatabihty and sample preparation at low concentration,by sample preparation and recovery at medium and high concentration.

Objective To investigate the effects of ginsenoside Rg3 (GS-Rg3) on the hypertrophic scars of rabbit ears and provide experimental foundation for study on its inhibition on the hypertrophic scars. Methods Hypertrophic scars were proved on 24 white rabbits,of which the whole level of the skin was excised for 2 cm?2 cm,4-6 points for each ear,controlled by itself. GS-Rg3 0.1 mL(concentration 3 g?L-1) was injected into experimental group and the same volume of saline solution into control group,once every three days regionally. The scar tissues were collected 2,4 and 6 weeks after the injection respectively,the thickness of the scar,structure under the microscope,and the expressions of PCNA,Bcl-2 and Bax were observed. Results In control group,three weeks after the epithelization of the wound,the thickness of the hypertrophic tissue was 3-4 times of ventro ear skin. Under microscope,the dermis was hyperplasia and got thicker,consisted with amount of fibroblast cells,collagen and vessels,the collagen was untidy,nodule or vortex,and the cartilage could be observed in some region.In experimental group, six weeks after the injection,the skin got thinner,the collagen became neath and the quantity of the vessels decreased. In the hypertrophic scars,there was high expression of PCNA,the percent of positive cells was higher (39.55%?6.07%) compared normal tissue (11.18%?1.71%).In GS-Rg3 group,the expression of Bcl-2 was gradually decreased two weeks after injection and obviously decreased six weeks later,there was significant difference compared with before injection (P