Effect of Nucleolin on Lymphoma Proliferation by Regulating Thymidine Kinase 1 / 中国实验血液学杂志

OBJECTIVE@#To investigate the mechanism of nucleolin (NCL) involved in lymphoma proliferation by regulating thymidine kinase 1 (TK1).@*METHODS@#Twenty-three patients with diffuse large B-cell lymphoma (DLBCL) were selected and divided into initial treatment group (14 cases) and relapsed/refractory group (9 cases). Serum TK1 and C23 protein in peripheral blood mononuclear cells were detected. Cell models of CA46-NCL-KD (CA46-NCL-knockdown) and CA46-NCL-KNC (CA46-NCL-knockdown negative control) were established by lentivirus vector mediated transfection in Burkitt lymphoma cell line CA46. The half maximal inhibitory concentration (IC50) of CA46-NCL-KD, CA46-NCL-KNC, and CA46 to adriamycin were detected by cell proliferation assay (MTS). The expression of NCL mRNA and protein in CA46-NCL-KD and CA46-NCL-KNC cells were dectected by Q-PCR and Western blot, respectively. The cell cycle of CA46-NCL-KD, CA46-NCL-KNC, and CA46 cells were detected by flow cytometry. The expression of TK1 protein in CA46-NCL-KD and CA46-NCL-KNC cells was detected by an enhanced chemiluminescence (ECL) dot blot assay.@*RESULTS@#The level of serum TK1 in the initial treatment group was 0.43(0-30-1.01) pmol/L, which was lower than 10.56(2.19-14.99) pmol/L in the relapsed/refractory group (P<0-01), and the relative expression level of NCL protein in peripheral blood was also significantly lower. The IC50 of CA46-C23-KD cells to adriamycin was (0.147±0.02) μg/ml, which was significantly lower than (0.301±0.04) μg/ml of CA46-C23-KNC cells and (0.338±0.05) μg/ml of CA46 cells (P<0.05). Compared with CA46-NCL-KNC cells, the expression of NCL mRNA and protein, TK1 protein decreased in CA46-NCL-KD cells, and the proportion of S phase and G2/M phase also decreased, while G0/G1 phase increased in cell cycle.@*CONCLUSION@#The increased expression of NCL in DLBCL and CA46 cells indicates low sensitivity to drug. NCL may participate in regulation of lymphoma proliferation by affecting TK1 expression, thereby affecting the drug sensitivity.

Incidence and influencing factors of parastomal hernia in patients with permanent colostomy / 中华消化外科杂志

Objective:To investigate the incidence and influencing factors of parastomal hernia in patients with permanent colostomy.Methods:The retrospective cohort study was conduc-ted. The clinicopathological data of 72 patients with permanent colostomy in the Beijing Friendship Hospital of Capital Medical University from January 2016 to June 2020 were collected. There were 50 males and 22 females, aged (66±12)years. Observations indicators: (1) follow-up; (2) analysis of factors affecting the incidence of parastomal hernia; (3) comparison of the incidence of parastomal hernia in patients with different age. Follow-up was conducted using outpatient examination. Patients were followed up once every 12 months after surgery to detect the incidence of parastomal hernia up to September 2021. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(range). Count data were expressed as absolute numbers and percentages, and comparison between groups was conducted using the chi-square test. Univariate analysis was conducted using the corresponding statistical methods based on data type. Multivariate analysis was conducted using the Logistic regression model. Kaplan-Meier method was used to draw the parastomal hernia occurrence curve and calculate the incidence rate of parastomal hernia and Log-rank test was used to analyze the incidence of parastomal hernia. Results:(1) Follow-up. All 72 patients were followed up for 23(range, 12?76)months. During the follow-up, there were 31 patients developed parastomal hernia, with the incidence as 20.8%(15/72), 36.1%(26/72) and 43.1%(31/72) at postoperative 1 year, postoperative 2 year and postoperative 5 year, respectively. Of the 31 patients with parastomal hernia, there were 21 cases of type Ⅰ, 3 cases of type Ⅱ and 7 cases of type Ⅲ. Patients with parastomal hernia recovered with conservative treatment. (2) Analysis of factors affecting the incidence of parastomal hernia. Results of univariate analysis showed that age, subcutaneous fat thickness and rectus abdominis thickness were related factors affecting the incidence of parastomal hernia ( χ2=7.98, t=?2.95, 2.02, P<0.05). Results of multivariate analysis showed that age, subcutaneous fat thickness and rectus abdominis thickness were independent factors affecting the incidence of parastomal hernia ( odds ratio=4.07, 3.19, 0.07, 95% confidence interval as 1.46?11.32, 1.43?7.09, 0.01?0.84, P<0.05). (3) Comparison of the incidence of parastomal hernia in patients with different age. Of the 72 patients, there were 37 cases with age <65 years and 35 cases with age >65 years. Of the 31 patients with parastomal hernia, there were 10 cases with age<65 years and all of them with type Ⅰ parastomal hernia, and the incidence of parastomal hernia in postoperative 1 year and postoperative 2 year was 13.5%(5/37) and 27.0%(10/37), respectively. There were 21 cases with age ≥65 years and cases with type Ⅰ, type Ⅱ and type Ⅲ parastomal hernia were 11, 3 and 7, respectively. The postoperative 1 year and postoperative 2 year incidence of parastomal hernia in the 21 cases was 28.6%(10/35) and 45.7%(16/35), respectively. There was a significant difference in the incidence of parastomal hernia between patients<65 years and ≥65 years ( χ2=9.28, P<0.05). Conclusion:Age, subcutaneous fat thickness and rectus abdominis thickness are independent factors affecting the incidence of parastomal hernia.

PLK1 Expression in Mantle Cell Lymphoma and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To explore the expression level of PLK1 in mantle cell lymphoma(MCL), and the effect of silencing PLK1 gene by RNA interference on the cell proliferation, apoptosis, and cell cycle.@*METHODS@#S-P immunohistochemistry technique was used to detect the expression of PLK1 in tissues of 42 patients with MCL and 30 patients with reactive proliferative lymphodenitis(RPL), their expression levels were compared and analyzed. The Jeko-1 cells were transfected with lentivirus contaiming PLK-1 shRNA, then the mRNA and protein expression of PLK-1 was detected by real-time guantitative PCR and Western blot nespectively, and the silencing efficacy of PLK-1 shRNA was identificd. The cell proliferation was detected by CCK method, the cell apoptosis was detected by Annexin V/PI double staining, the cell cycle was detected by PI single staining, the changes of apoptosis-related proteins BAX, BCL-2 and Caspase 3 were detected by Western blot.@*RESULTS@#The positive expression rate of PLK-1 in tissue of MCL patients was 66.67%(28/42), which was significanfly higher than 20%(6/30) in tissue of RPL patients (P＜0.05). The PLK-1 positive expression correlated with B symptom, IPI score, Ann-Arbor stage(P＜0.05). After infection of Jeko-1 cells with lentivirus containing PLK-1 shRNA for 72 hours, the mRNA and protein expressions of PLK-1 were significantly down-regulated(P＜0.05), the proliferation rate of cells in group of PLK-1 shRNA was significanly lower than that in control and Neg shRNA groups(P＜0.05); the apoptosis rate of cells in PLK-1 shRNA group was (27.42±3.44)%, which was significantly higher than that in control group (1.23±0.42)% and Neg shRNA group (2.07±0.58) % (P＜0.05). The cell cycle analysis showed that the cell ratio in G/M phase of PLK-1 shRNA group was (27.21±3.59) %, which was higher than that in control group (13.28±2.63)% and Neg shRNA group (14.34±2.37) %. The detection of apoptosis-related proteins showed that the expression of BAX was up-regulated, the expression of BCL-2 was down-regnlated and the expression of caspase 3 was up-regulated.@*CONCLUSION@#The PLK-l overexpression appears in tissue of MCL patients. The silencing PLK-1 gene can inhibit the proliferation of Jeko-1 cells, induce the apopotosis of Jeko-1 cells and arrestes cell cycle in G/M phase.

The analysis of the sacroiliac joint CT examination in the detection rate and missed diagnosis of sacral canal cyst / 中国医师进修杂志

Objective To investigate the detection rate and missed diagnosis reason of sacral canal cyst with sacroiliac joint CT scan.Methods Retrospectively analyzed the changes of sacral canal in 1 286 cases with sacroiliac joint CT scan.CT scanning included bone algorithm reconstruction and standard algorithm reconstruction.Results Among 1 025 patients who had negative sacroiliac joint CT scan were found in 36 cases of sacral canal cyst,the detection rate was 3.5％ (36/1 025).Single cyst in 34 cases and multiple cysts in 2 cases.Cyst in S1 level 5 cases,S1-2 level 4 cases,S2 level 22 cases,S2-3 level 4 cases,S3 level 3 cases.The minimal short diameter of the cysts was 0.5 cm and the maximal diameter was 3.2 cm,average 1.2 cm.CT scan showed a round uniform cystic low density,and combined with sacral canal expansion in 19 cases.CT scan used bone algorithm reconstruction in 29 cases,used standard and bone algorithm reconstruction in 7 cases.Clinical findings and correctly diagnosed in 11 cases,accounted for 30.6％ (11/36),about 69.4％ (25/36) cases were missed diagnosis.Conclusions About 3.5％ patients are found sacral canal cyst who have negative sacroiliac joint CT scan.Both the standard and bone algorithm reconstruction should be used to avoid the missed diagnosis of sacral canal cyst.

Chemical constituents and pharmacologic actions of Cynomorium plants / 中国天然药物

The stem of Cynomorium songaricum is a traditional Chinese medicine reputed to have tonic effects. C. coccineum growing in northern Africa and the Mediterranean region is regarded in Arabian medical practice as the "treasure of drugs". The major constituents of Cynomorium plants have been revealed to be phenolic compounds, steroids, triterpenes, etc. Pharmacologic studies showed that the Cynomorium plants had antioxidant, immunity-improving, anti-diabetic, neuroprotective, and other bioactivities. Some chemical constituents in Cynomorium plants are unstable, implying that the chemical components of the herbal medicines produced under different conditions may be variable. This review covers the literature published until December, 2011 and describes the pharmacologic effects and secondary metabolites of Cynomorium species.

Influence of different scoring methods on preventive anticoagulant therapy after risk stratification in atrial fibrillation patients / 中国基层医药

Objective To explore which scoring methods can prevent the ischemic stroke better.Methods 80 padents with atrial fibrillation were randomly divided into two groups,the patients in A group took anticoagulant therapy using CHADS2 score;the patients in B group took anticoagulant therapy using CHADS2-VAS score;follow-up the ischemic stnoke,death and major bleeding event rate.Results 7 cases of ischemic stroke in A group,1 case in B group,the difference was statistically significant ( P ＜ 0.05 ).Conclusion According to CHA2DS2-VAS score for risk stratification,anticoagulant therapy was safe and effective,and did not increase bleeding event rate.

Investigation on the mechanisms of p15INK4B gene demethylation by valproate in Molt-4 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the antitumour effects of sodium valproate (VPA) on the proliferation, differentiation and cell cycle of Molt-4 cell and to investigate its demethylation mechanisms.</p><p><b>METHODS</b>After Molt-4 cells trated with VPA at different concentrations, cell viability and growth curve were assessed by MTT assay. Cell cycle changes were analyzed by flow cytometry. The expression level of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR.</p><p><b>RESULTS</b>VPA significantly inhibited the proliferation of Molt-4 cells. After 48 h culture with 5.0 mmol/L VPA, the percentages of Molt-4 cells in G(0)/G(1) phase was (66.87 ± 3.31)% and in S phase was (8.47 ± 2.56)%, while in control group, the cells in G(0)/G(1) phase increased and in S phase decreased significantly. The p15 gene in Molt-4 cells failed to express due to its hypermethylation. The expression level of p15 gene mRNA increased significantly after exposure to VPA for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner. The expression level of DNMT3B decreased at 10.0 mmol/L concentration.</p><p><b>CONCLUSION</b>VPA has a demethylation effect on p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities to recover p15 gene activity, which arrests Molt-4 cell in G(0)/G(1) phase.</p>

Inhibitory effect of VPA on multiple myeloma U266 cell proliferation and regulation of histone acetylation / 中国实验血液学杂志

This study was aimed to investigate the effects of sodium valproate (VPA) on the proliferation and regulation of histone acetylation of multiple myeloma cell line U266. U266 cells were treated with VPA. Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). The expression level of HDAC1 mRNA was detected by RT-PCR, and the protein levels of HDAC1 and histone H3, H4 acetylation was detected by Western blot. The results showed that the VPA inhibited the proliferation of U266 cells in concentration-and time-dependent manners.After exposure to different concentrations of VPA for 48 hours, the proportion of G(0)/G(1) cells increased, while the proportion of S phase cells decreased. The cell cycle was arrested obviously in G(0)/G(1) phase (p < 0.05). The expression of HDAC1 mRNA was inhibited, and the protein level of HDAC1 was down-regulated, while the histone H3/H4 acetylation was up-regulated in U266 cells. It is concluded that the VPA can inhibit cell proliferation of U266 and induce G(0)/G(1) phase arrest. The increase of histone H3/H4 acetylation resulting from inhibiting HDAC1 by VPA might be considered as a possible mechanism.

Synergistic effects of VPA and As2O3 on Molt-4 cells in vitro and its possible mechanisms / 中国实验血液学杂志

This study was purposed to investigate the synergistic effects of sodium valproate (VPA) and As2O3 on the proliferation of Molt-4 cells in vitro and its possible mechanisms. Cell viability and growth curve were assessed by the MTT assay. The synergistic activity in combination of 2 drugs was determined by the Q format. The expression levels of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and DNMT 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR. The results indicated that the VPA and As2O3 both inhibited proliferation of Molt-4 cells. The combination of two drugs showed an additive effect (values of Q were>or=0.85). The inhibitory rate in combination of 5 mmol/L of VPA with 10 micromol/L of As2O3 was (70.31+/-2.54)%. The p15 gene in Molt-4 cell line failed to express due to its hypermethylation. The level of p15 gene mRNA expression increased significantly after exposure to VPA in combination with As2O3 for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner, whereas DNMT3A had no significant differences from the control. The level of expression of DNMT3B seemed to decrease at 10 mmol/L concentration. There were significant differences between the combination of the two drugs and the control group. The gray value of methylated bands decreased after the treatment of VPA alone and in combination with As2O3 in a dose-dependent manner. It is concluded that VPA induces demethylation of p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities, which up-regulates the p15 gene, recovers its activity. The combination of VPA with As2O3 has the synergistic additive effect on the inhibition of cell viability, so that VPA can reduce the side effect of As2O3 on liver function, which would be verified in the clinical practice.

Effect of wolfberry fruit and epimedium on DNA synthesis of the aging-youth 2BS fusion cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of water extracts of Wolfberry fruit (WB) and Epimedium (EM) on DNA synthesis of the aging-youth 2BS fusion cells.</p><p><b>METHODS</b>Human embryonic lung diploid fibroblasts 2BS national standard strain, were used as an aging model. Cell denucleation and cell fusion techniques were applied to observe the effect of WB and EM on DNA synthesis of 2BS fusion cells.</p><p><b>RESULTS</b>In the 0.025 (V/V) WB or EM water extract containing media, 2BS cells could be continuously cultured for 61.0 +/- 2.9 passages and 56.0 +/- 2.6 passages respectively, while in the control group it was only 49.0 +/- 2.6 passages (P < 0.01). After treatment with WB and EM separately for 2 hrs, the aging 2BS cells were denucleated and fused with young 2BS cells. The [3H]TdR incorporation percentage in these treated cells was significantly higher than that in the untreated control cells (P < 0.01).</p><p><b>CONCLUSION</b>Both WB and EM can accelerate the DNA synthesis rate of the aging youth 2BS fusion cells and prolong the life span of 2BS cells.</p>