OBJECTIVE: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of daidzein, daidzin, genistein and genistin in Semen Sojae Praeparatum.

Glycerol is the main byproduct in ethanol production by Saccharomyces cerevisiae. In order to improve ethanol yield and the substrate conversion, a cassette about 4.5 kb for gene homologous recombination, gpd2Δ::PGK1(PT)-POS5-HyBR, was constructed and transformed into the haploid strain S. cerevisiae S1 (MATa) to replace the GPD2 gene by POS5 gene. The NADH kinase gene POS5 was successfully over expressed in the recombinant strain S. cerevisiae S3. Comparing with the parent strain, the recombinant strain S. cerevisiae S3 exhibited an 8% increase in ethanol production and a 33.64% decrease in glycerol production in the conical flask fermentation with an initiatory glucose concentration of 150 g/L. Overexpression of NADH kinase gene seems effective in reducing glycerol production and increasing ethanol yield.
Ethanol ; chemistry ; Fermentation ; Glycerol ; chemistry ; Industrial Microbiology ; Mitochondrial Proteins ; genetics ; metabolism ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; metabolism

Based on collections and researches of Pesudostellaria heterophylla germplasm resources from different areas of China, by using Shibing SB-4 provenance as materials, the new variety "Shitai No.1" was bred by mass selection, small plot variety comparative test, regional variety comparative test and field trial planting. Compared with "Qian taizishen No.1" and P. heterophylla land races. The disease and lodging resistance, root yield, polysaccharide content and the first grade rate of "Shitai No.1" have obvious advantages. In addition, it is relatively stable of yield in "Shitai No.1" in different places. It is demonstrated that "Shitai No.1" is a fine variety that adapt to the producing areas of P. heterophylla in Guizhou province, it is worthy to be promoted.

OBJECTIVE：To estab lish fingerprint of Duzhong butiansu pill s，analyze its chemical pattern recognition ，and determine the contents of 7 components in Duzhong butiansu pills ，so as to provide reference for the quality control of the preparation. METHODS ：HPLC method was adopted. The determination was performed on Pntulips BP-C 18 Plus column with 0.2％ phosphoric acid water-acetonitrile as mobile phase （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 330 nm，and column temperature was 35 ℃. The sample size was 20 μL. With paeonol as the reference，the HPLC fingerprints of 12 batches of Duzhong butiansu pills （S1-S12） were established with Similarity Evaluation System for TCM Chromatographic Fingerprint （2012 edition）； common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference substance. SPSS 21.0 and SIMCA 13.0 software were used for cluster analysis and principal component analysis ，and 22 common peaks were evaluated. The contents of the identified components in 12 batches of samples was determined by the above HPLC method. RESULTS ：A total of 22 common peaks were identified in the HPLC fingerprint of 12 batches of Duzhong butiansu pills ，and the similarity was no loss than 0.960. There were 7 chemical components identified ，which were gallic acid （peak 1），chlorogenic acid （peak 3），liquiritoside（peak 6），hyperoside （peak 7），verbascoside（peak 8），icariin（peak 14）and paeonol （peak 15）. Among the 12 batches of samples ，S1，S3-S5，S7， S9 and S 11 were classified as one category ，S2，S10 and S 124Y091 were clustered into one category ，S6 was one category and S was one category. The 22 common peaks were divided into three principal components. The characteristic value （15.130） and contribution rate （68.775％） of principal component 1 were the largest ，and the score coefficients of peak 3（0.305）and peak 4（0.298）were the highest. Among 12 batches of samples，the cont ents of above 7 components were 18.196 231.951 3，0.000 6-0.049 4，0.234 8-0.415 9，0.039 5-0.079 1，0.053 5-0.249 3，0.000 5-0.000 8，0.646 4-1.146 9 mg/g，respectively. CONCLUSIONS：HPLC fingerprint of Duzhong butiansu pills is established successfully. Twelve batches of samples are clustered into 4 category. Peak 3（chlorogenic acid ）and peak 4（unknown）may be the important factors causing the difference of samples. The content of gallic acid is the highest among the 7 components.

Alcoholic Beverages ; adverse effects ; Animals ; Carcinoma, Hepatocellular ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Suppressor Protein p53 ; metabolism

BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.

METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.

RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.

CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.

Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction

Objective: To discuss the safety and effectiveness of the improved technique by comparing the effects of low temperature bone cement infusion before and after the improvement in the percutaneous vertebroplasty (PVP). Methods: The clinical data of 170 patients (184 vertebrae) with osteoporotic vertebral compression fracture who met the selection criteria between January 2016 and January 2018 were retrospectively analyzed. All patients were treated with PVP by low-temperature bone cement perfusion technology. According to the technical improvement or not, the patients were divided into two groups: the group before the technical improvement (group A, 95 cases) and the group after the technical improvement (group B, 75 cases). In group A, the patients were treated by keeping the temperature of bone cement at 0℃ and parallel puncture; in group B, the patients were treated by increasing the temperature of bone cement or reducing the time of bone cement in ice salt water and cross puncture. There was no significant difference in gender, age, disease duration, T value of bone mineral density, operative segment, and preoperative vertebral compression rate, visual analogue scale (VAS) score between the two groups ( P>0.05). CT examination was performed immediately after operation, and the leakage rate of bone cement was calculated. The amount of bone cement perfusion and the proportion of bone cement in contact with the upper and lower endplates at the same time were compared between the two groups. The vertebral compression rate was calculated and the VAS score was used to evaluate the pain before operation, at immediate after operation, and last follow-up. Results: There was no complication such as incision infection, spinal nerve injury, or pulmonary embolism in both groups. There was no significant difference in the amount of bone cement perfusion between groups A and B ( t=0.175, P=0.861). There were 38 vertebral bodies (36.89%) in group A and 49 vertebral bodies (60.49%) in group B exposed to bone cement contacting with the upper and lower endplates at the same time, showing significant difference ( χ2=10.132, P=0.001). Bone cement leakage occurred in 19 vertebral bodies (18.45%) in group A and 6 vertebral bodies (7.41%) in group B, also showing significant difference ( χ2=4.706, P=0.030). The patients in group A and group B were followed up (13.3±1.2) months and (11.5±1.1) months, respectively. The vertebral compression rates of the two groups at immediate after operation were significantly lower than those before operation ( P<0.05), but the vertebral compression rate of group A at last follow-up was significantly higher than that at immediate after operation ( P<0.05), and there was no significant difference in group B between at immediate after operation and at last follow-up ( P>0.05). The VAS scores of the two groups at immediate after operation were significantly lower than those before operation ( P<0.05); but the VAS scores of group A at last follow-up were significantly higher than those at immediate after operation ( P<0.05) and there was no siginificant difference in group B ( P>0.05). There was no significant difference in VAS scores between the two groups at immediate after operation ( t=0.380, P=0.705); but at last follow-up, VAS score in group B was significantly lower than that in group A ( t=3.627, P=0.000). Conclusion: The improved advanced low-temperature bone cement perfusion technology during PVP by increasing the viscosity of bone cement combined with cross-puncture technology, can reduce bone cement leakage, improve the distribution of bone cement in the vertebral body, and reduce the risk of vertebral collapse, and achieve better effectiveness.

AIM: To evaluate the effects of visual perceptual learning on visual representation and binocular vision function of children with amblyopia.

METHODS: A total of 94 cases(94 eyes)children with amblyopia admitted to the ophthalmology clinic of our hospital from January 2014 to December 2015 were selected for the study and were grouped according to the degrees of amblyopia, and they were given visual perceptual learning for 3mo. The clinical efficacy and perceptual level of eye position and poor visual acuity before and after treatment were compared among groups, and Pearson correlation coefficient was used to analyze the relationship between difference of poor visual acuity and difference of perceptual eye position before and after treatment in children with amblyopia.

RESULTS: The total clinical effective rate of children with mild amblyopia was higher than that of children with moderate or severe amblyopia(P<0.05), and there was no significant difference in the total clinical effective rate between children with moderate amblyopia and children with severe amblyopia(P>0.05). The perception level of eye position after treatment of children with amblyopia was significantly lower than that before treatment(P<0.05). The perception level of eye position after treatment in children with mild, moderate or severe amblyopia was significantly decreased(P<0.05), and the improvement of perception level of eye position in children with mild or moderate amblyopia was more significant than that in children with severe amblyopia(P<0.05). The poor visual acuity of children with amblyopia after treatment was significantly improved compared with that before treatment(P<0.05). The poor visual acuity after treatment in children with mild, moderate or severe amblyopia was significantly improvement(P<0.05), and the improvement of poor visual acuity in children with mild or moderate amblyopia was more significant than that in children with severe amblyopia(P<0.05). The difference of poor visual acuity was positively correlated with difference of perceptual eye position before and after treatment among children with amblyopia(r=0.371, P<0.05).

CONCLUSION: Visual perceptual learning can effectively improve the visual representation and binocular vision function of children with amblyopia, and it is suitable for the clinical treatment of children with amblyopia.

OBJECTIVETo investigate the clinical significance of p16 protein non-expression and p16 gene inactivation by deletions and hypermethylation in nasopharyngeal carcinoma.

METHODSImmunohistochemical study for p16 protein was carried out in 90 cases of non-keratinizing carcinoma (NKC) of nasopharynx. P16 gene deletions and hypermethylation were also analyzed by polymerase chain reaction (PCR) and methylation-specific PCR in 23 randomly selected NKC cases.

RESULTSAmong the 90 NKC cases studied, 42 cases (46.7%) were negative for p16 protein. The non-expression rate of p16 protein also correlated with the 5-year survival rate. The non-expression rate was 60.0% in patients who died within 5 years, in contrast to 20.0% in those alive for over 5 years after diagnosis. The non-expression rate of p16 protein in cases with or without distant metastasis was 81.8% and 41.8% respectively (P < 0.05), while that in cases with or without local invasion into skull base was 41.7% and 48.5% respectively (P > 0.05). As for molecular analysis, deletion of p16 gene exon 2 was found in 10 of the 23 cases (43.4%) studied, while deletion of p16 gene exon 1 was not detected in these samples. Hypermethylation of p16 gene exon 1 was also noted in 2 of the 23 cases (8.7%). The overall mutation rate of these cases was 52.1%.

CONCLUSIONSThere is a high incidence of p16 protein non-expression, deletion of p16 gene exon 2 and hypermethylation of p16 gene exon 1 in NKC. P16 gene inactivation may thus play an important role in the pathogenesis of NKC, especially in terms of its metastatic potential.

CpG Islands ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; DNA Methylation ; DNA, Neoplasm ; genetics ; Exons ; genetics ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Skull Base Neoplasms ; pathology ; Survival Rate

OBJECTIVETo discuss the value of P53mt and BCL-2 proteins in screening skin carcinoma due to arseniasis.

METHODSP53mt and BCL-2 proteins were detected by immunohistochemical staining. chi(2) test was used to analyze the difference of positive rate between two groups. Screening value of the two biomarkers was also evaluated through the analysis of relative indexes.

RESULTSPositive percentages of P53mt and BCL-2 in carcinoma group were 88.89% and 94.44% respectively, both were higher than those of 36.0% and 66.0% in non-carcinoma group (for P53mt, P < 0.01; for BCL-2, P < 0.05). ORs of P53mt and BCL-2 were 14.22 (2.93 - 68.97) and 8.76 (1.07 - 71.51), respectively. Youden's Index and specificity of P53mt were 0.529 and 64.0%, which were much higher than those of BCL-2. Serial tests improved the value of screening with Youden's Index 0.569, but parallel test lowered it to 0.244.

CONCLUSIONSP53mt and BCL-2 were practical biomarkers to screen skin carcinoma due to arseniasis, and the former was better than the latter. The value of screening can be improved by a series of tests.

Arsenic Poisoning ; complications ; Humans ; Immunohistochemistry ; Mass Screening ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Skin ; chemistry ; pathology ; Skin Neoplasms ; diagnosis ; etiology ; metabolism ; Tumor Suppressor Protein p53 ; metabolism