Objective:To investigate the expression of Cyclin D 1?p16?p27 and PCNA in glioma with different pathological classification, and to explore the correlationship among Cyclin D 1?p16?p27 and PCNA.Methods:Cyclin D 1?p16?p27 and PCNA protein expressions in 12 normal brain tissues and 58 glioma cases were examined by immunohistochemical staining and hemiquantity analysis.Results:With the increasing of glioma malignancy, the expression level of Cyclin D 1 protein increased gradually (P

OBJECTIVE:To establish a method for simultaneous determination of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate in Xiaoer huadu powder.METHODS:HPLC method was adopted.The separation was performed on Waters SunFireTM-C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 238 nm,and column temperature was 25 ℃.The sample size was 10 μL.RESULTS:The linear ranges of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate were 8.808-88.08 μg/mL(r=0.999 8),1.778-17.78 μg/mL(r=0.999 6),2.533-25.33 μg/mL(r=0.999 9),respectively.LOQ were 4.404,0.889,2.533 μg/mL;LOD were 1.101,0.445,1.267 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 95.08％-99.61％ (RSD=1.77％,n =9),96.93％-99.94％ (RSD=0.92％,n=9),98.33％-102.05％ (RSD=1.27％,n=9).CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for simultaneous determination of paeoniflorin,berberine hydrochloride and ammonium glycyrrhizinate in Xiaoer huadu powder.

OBJECTIVE:To observe clinical efficacy and safety of alprostadil combined with salvia ligustrazine in the treat-ment of aged patrents with unstable angina pectoris. METHODS:A total of 150 patients with unstable angina pectoris department of our hospital during Oct. 2011-Mar. 2015 were randomly divided into alprostadil group,salvia ligustrazine group and combination group according to random number table,with 50 cases in each group. Three groups received routine treatment. Alprostadil group additionally received Alprostadil injection 100 μg added into 0.9％ Sodium chloride injection 250 mL,ivgtt,qd,on the basis of routine treatment. Salvia ligustrazine group additionally received Salvia ligustrazine injection 10 mL added into 0.9％ Sodium chlo-ride injection 250 mL,ivgtt,qd,on the basis of routine treatment. Combination group additionally received constant dose of Al-prostadil injection and Salvia ligustrazine injection. Hemorheological indexes (high shear whole blood viscosity,low shear whole blood viscosity,plasma viscosity,hematocrit,fibrinogen),cardiac function indexes(LVEF,SV,LVEFD,LVST),serum CRP, NO,ET,SOD and clinical efficacies were observed in 2 groups before and after treatment;the occurrence of ADR was compared between 2 groups. RESULTS:Before treatment,there was no statistical significance in hemorheological indexes,cardiac function indexes or serum CRP,NO,ET,SOD level between 2 groups (P>0.05). After treatment,plasma viscosity,the whole blood high and low shear viscosity,hematocrit,fibrinogen,serum CRP and ET levels of 3 groups were decreased significantly,while LVEF,SV,serum levels of NO and SOD were increased significantly,combination group was significantly better than alprostadil group and salvia ligustrazine group,with statistical significance (P<0.05). There was no statistical significance in above indexes between alprostadil group and salvia ligustrazine group (P<0.05). Total response rate of combination group was 90.00％,which was significantly higher than 74.00％of alprostadil group and 72.00％of salvia ligustrazine group,with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups (P>0.05). CONCLUSIONS:Alprostadil combined with salvia ligustrazine can effectively reduce the blood viscosity of patients with unstable angina pectoris,improve cardi-ac function and endothelial function,reduce myocardial ischemia injury and show significant therapeutic efficacy and safety without increasing the incidence of ADR.

Objective: To establish the quality standard for Qiankun Liquid.Methods: Fructus Lycii、Fructus Cnidii、Radix Ginseng in Qiankun Liquid were identified by TLC, and the content of icariin was determined by HPLC.Results: Fructus Lycii、Fructus Cnidii、Radix Ginseng could be identified by TLC. Icariin showed a good linear relationship at a range of 0.126～1.008?g, r =0.9999. Conclusion: The method is accurate and can be used for the quality control of Qiankun Liquid.

OBJECTIVE:To observe the effects of Gegenqinlian colon positioning tablet(GGQLJC)on colon tissue PPAR-γ, NF-κB p65 protein expressions of model rabbits with damp-heat type ulcerative colitis(UC). METHODS:56 rabbits were random-ly divided into normal group(normal saline),model group(normal saline),sulfasalazine tablet(SASP)group(positive control, 0.300 g/kg),Gegenqinlian tablet (GGQL) group (0.225 g/kg) and GGQLJC high-dose,medium-dose,low-dose groups (1.036, 0.518,0.259 g/kg),8 in each group. Except for normal group,rabbits in other groups were cultured for damp-heat-type UC mod-el,intragastrically administrated in the second day of last administration,once a day,for 14 d. Disease activity index(DAI),co-lonic mucosal damage index (CMDI),histological damage (TDI) were scored;colon,spleen and thymus indexes were deter-mined;PPAR-γ,NF-κB p65 protein expressions in colon tissue were detected. RESULTS:Compared with normal group,DAI, CMDI,TDI scores and spleen index,colon index,NF-κB p65 protein level in colon tissue in model group were significantly in-creased(P<0.01);thymus index,PPAR-γprotein level in colon tissue were significantly reduced(P<0.01). Compared with mod-el group,above-mentioned indexes in each administration group were significantly improved (P<0.05 or P<0.01). Compared with GGQL group,DAI and TDI scores,spleen index,colon index,NF-κB p65 protein level in colon tissue in SASP group, GGQLJC high-dose,medium-dose groups were significantly decreased (P<0.05);PPAR-γ protein level in colon tissue in SASP group,GGQLJC high-dose,medium-dose groups were significantly increased (P<0.05 or P<0.01). CONCLUSIONS:GGQLJC has certain improvement effects on model rabbits with damp-heat type UC,which is superior to GGQL. The mechanism may be re-lated to increasing PPAR-γprotein level and decreasing NF-κB p65 protein level in colon tissue.

OBJECTIVE:To establish a method for simultaneous determination of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride in Shenbai shuxin granules.METHODS:HPLC method was adopted.The separation was performed on Waters sunfire-C18 column with mobile phase consisted of acetonitrile-0.05 ％ trifluoroacetic acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 288 nm,and the column temperature was 30 ℃.The sample size was 5 μL.RESULTS:The linear ranges of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride were 0.040 01-1.600 46 μg(r=0.999 9),0.013 84-0.553 7 μg(r=0.999 9),0.049 32-1.972 94 μg(r=0.999 6),0.014 46-0.578 6 μg(r=0.999 8).The limits of quantitation were 7.68,2.66,4.74,1.38 ng,and the limits of detection were 1.92,0.66,2.36,0.69 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 98.846％-100.762％ (RSD=0.77％,n=6),96.632％-99.463％ (RSD=0.98％,n=6),98.541％-100.432％ (RSD=0.82 ％,n =6),98.607 ％-101.521％ (RSD =1.11％,n =6),respectively.CONCLUSIONS:The method is simple,accurate and precise.It can be used for 4 components in Shenbai shuxin granules.

Objective The potential mechanism of hepatotoxicity induced by rhubarb was preliminarily explored by network pharmacology and verified by cell experiments.Methods Based on network pharmacology,component collection and target prediction are carried out through multiple databases.PPI network construction,GO enrichment analysis and KEGG pathway analysis were combined with software to systematically predict the mechanism of hepatotoxicity induced by rhubarb.The pathway information predicted by network pharmacology was verified by primary hepatocyte experiments and Western blot experiments.Results The results of network pharmacology showed that RH was the main component of hepatotoxicity induced by rhubarb.Seventeen core targets of hepatotoxicity induced by rhubarb were obtained.KEGG results suggested that DNA damage and apoptosis were one of the key mechanisms of hepatotoxicity induced by rhubarb.The results of primary hepatocytes and Western blot showed that RH could inhibit the viability of primary hepatocytes in a time-dose dependent manner.ABT and SFP can significantly reduce the toxicity of RH on primary liver cells in mice,and RFP can increase the toxicity of RH to mouse primary liver cells.Upregulation of γ-H2AX and PARP-1 protein in primary liver cells of mice after treatment with different concentrations of RH.Conclusion RH in rhubarb can significantly inhibit the viability of mouse primary hepatocytes,and its toxicity to mouse primary hepatocytes is mainly caused by the metabolic activation of RH by CYP 2C9.RH can activate PARP-1 protein,phosphorylate H2AX,induce DNA damage and apoptosis in mouse primary hepatocytes.

OBJECTIVETo study the pattern of CGG repeat instability within germline cells derived from two male fetuses affected with Fragile X syndrome (FXS).

METHODSThe length and methylation status of CGG repeats within the testes of a fetus carrying a full FXS mutation and another fetus carrying mosaicism FXS mutation were analyzed with Southern blotting and AmplideX FMR1 PCR. Immunohistochemistry was also applied for the measurement of FMR1 protein (FMRP) expression within the testes.

RESULTSFor the fetus carrying the full mutation, Southern blotting analysis of the PCR product has detected an expected band representing the full mutation in its brain and a premutation band of > 160 CGG repeats in its testis. Whereas the pattern of premutation/full mutation in mosaic testis was similar to that in peripheral blood and no sign of contracted fragment was found other than a band of about 160 CGG repeats. Immunohistochemistry assay with a FMRP-specific antibody demonstrated a number of FMRP-positive germ cells, which suggested a contraction from full mutation to premutation alleles.

CONCLUSIONThis study has clarified the instability pattern of CGG repeat and expression of FMRP protein within the testes of fetuses affected with FXS, confirming that the CGG repeat can contract progressively within the germline. The FMRP expression in the testis is consistent with spermatogonium proliferation, and thus the contraction from full mutation to unmethylated premutations may occur for the requirement of FMRP expression during spermatogenesis. The better understanding of FMRP function during germ cell proliferation may elucidate the mechanism underlying the contraction of full FXS mutation in male germline.

Abortion, Eugenic ; Blotting, Southern ; Brain ; embryology ; metabolism ; DNA Methylation ; Fatal Outcome ; Fetus ; cytology ; metabolism ; Fragile X Mental Retardation Protein ; genetics ; metabolism ; Fragile X Syndrome ; diagnosis ; genetics ; Humans ; Immunohistochemistry ; Male ; Mosaicism ; Mutation ; Polymerase Chain Reaction ; Spermatozoa ; metabolism ; Testis ; cytology ; embryology ; metabolism ; Trinucleotide Repeat Expansion ; genetics

Objective:To explore the efficacy and safety of in-class transition from proteasome inhibitor bortezomib to ixazomib in the treatment of newly-treated patients with multiple myeloma (MM).Methods:The clinical data of 63 newly-treated MM patients in Shenzhen Second People's Hospital from January 2018 to December 2020 were retrospectively analyzed. They were divided into transition group (23 cases) and bortezomib group (40 cases). Both groups were treated with bortezomib-containing regimen as the first-line treatment regimen. In case of intolerable adverse reactions, patients in the transition group were treated with ixazomib instead of bortezomib, while the patients in the bortezomib group did not undergo drug transition. The curative effect and progression-free survival (PFS) were compared between the two groups.Results:In the transition group, the overall response rate (ORR) before in-class transition was 95.7% (22/23), the rate of ≥ very good partial remission (VGPR) was 52.2% (12/23); the ORR after transition was 95.7% (22/23), and the rate of ≥ VGPR was 82.6% (19/23). In the bortezomib group, ORR was 90.0% (36/40), and the rate of ≥ VGPR was 72.5% (29/40). There was no significant difference in ORR and the rate of ≥VGPR between the two groups ( χ2 = 0.64, P=0.424; χ2 = 0.82, P = 0.364). The median number of cycles of PI therapy in the transition group was 9, and the median PFS time was not reached. The median number of cycles of PI therapy in the bortezomib group was 7.5, and the median PFS time was 30.0 months (95% CI 19.1-40.9 months), there was no significant difference in PFS between the two groups ( P = 0.275). In the bortezomib group, 12 patients discontinued bortezomib due to adverse reactions, the median PFS time was 20.0 months (95% CI 12.6-27.4 months), and the PFS of patients who discontinued PI in the transition group and the bortezomib group was compared, the difference was statistically significant ( P = 0.043). In the transition group, 21 patients (21/23, 91.3%) developed peripheral neuropathy, and the incidence of ≥grade 3 adverse reactions was 13.0% (3/23); in the bortezomib group, 22 patients (22/40, 55.0%) developed peripheral neuropathy, and the incidence of ≥grade 3 adverse reactions was 12.5% (5/40). Conclusions:For newly-treated MM patients, the transition from bortezomib to ixazomib can improve the depth of remission and reduce the recurrence caused by the discontinuation of PI.

Objective To compare the CGG-repeat-length and its methylation status in fetal tissues and to explicate the heterogeneity of CGG repeats.Method Multiple tissues from a full mutation (August 2013) and a mosaic aborted fetus of 23-week gestation(May 2012) were collected and genomic DNA from these tissues was extracted.The CGG-repeat-length and methylation status in fetal tissues were determined by a combined strategy of Southern blotting and GC-Rich PCR.FMR1 expression was measured by real time PCR and Western blotting.Result CGG-repeat-length in different tissues of each fetus was similar.A major methylated band in the full mutation range (540 CGG repeats) was detected in the brain,skin,testis and kidney tissues of Case 1.An unmethylated premutation band with 160 CGG repeats,and another two bands with 470 and 1 100 CGG repeats in the full mutation range were shown in the brain,skin,testis,lung,stomach,gut,liver,kidney,heart and blood of Case 2.However,the methylation status of CGG repeats in the mosaic fetus was heterogeneous among different tissues.The lowest premutation ratio was in the brain of the mosaic fetus compared with other tissues,and correspondingly FMR1 expression in its brain was minimum.Conclusion This study clarify the tissue heterogeneity of CGG repeats and provides information for the genetic counseling and clinical diagnosis in fragile X syndrome.Based on the fact that the mosaic fetus' mother is a carrier of full mutation,it is speculated that the maternal CGG repeat has contracted before the differentiation of trilaminar germ disc.