Mechanism of circZNF609 targeting miR-153 to regulate the proliferation and apoptosis of diffuse large B-cell lymphoma / 中华肿瘤杂志

Objective: To investigate the molecular mechanism of circZNF609 targeting miR-153 to regulate the proliferation and apoptosis of diffuse large B-cell lymphoma. Methods: Fifty cases of lymphoma tissue from patients with diffuse large B-cell lymphoma who were diagnosed and treated in the First Affiliated Hospital of Zhengzhou University from July 2018 to December 2019 were collected. Thirty cases of normal lymph node tissues that were confirmed to be reactive hyperplasia by pathological diagnosis during the same period were selected as controls. Real time quantitative polymerase chain reaction (PCR) was used to detect the expression of circZNF609 in diffuse large B-cell lymphoma tissues and control hyperplasia lymph nodes. Diffuse large B-cell lymphoma OCI-LY19 cells were divided into control group (blank control), si-con group (transfected with siRNA control), si-ZNF609 group (transfected with circZNF609 siRNA), and si-ZNF609+ Anti-NC group (co-transfected with circZNF609 siRNA and inhibitor control) and si-ZNF609+ Anti-miR-153 group (co-transfected with circZNF609 siRNA and miR-153 inhibitor). Cell counting kit-8 (CCK-8) was used to detected proliferation, flow cytometry was used to detect cell cycle and apoptosis. Western blot was used to detect the protein expressions of C-caspase-3, cyclin D1, p21. The luciferase reporter system was used to identifie the relationship between circZNF609 and miR-153. Results: The expression level of circZNF609 in diffuse large B-cell lymphoma tissue was (1.44±0.22), higher than (0.37±0.14) in the control tissues (P<0.001). The cell survival rate of the si-ZNF609 group was (51.74±6.39)%, lower than (100.00±10.23)% of the control group and the (99.64±11.67)% of the si-con group (P<0.001). The proportion of cells in the G(0)/G(1) phase was (63.25±4.11)%, higher than (48.62±4.32)% of the control group and (47.12±3.20)% of the si-con group (P<0.001), the apoptosis rate was (13.36±1.42)%, higher than (3.65±0.47)% of the control group and (3.84±0.62)% of the si-con group (P<0.05). The expression levels of C-caspase-3 and p21 protein were (0.85±0.09) and (0.90±0.08), higher than (0.38±0.04) and (0.65±0.07) in the control group and (0.39±0.05) and (0.66±0.05) in the si-con group (P<0.001). The expression level of cyclin D1 protein was (0.40±0.03), lower than (0.52±0.06) of the control group and (0.53±0.04) of the si-con group (all P<0.001). CircZNF609 and miR-153 are mutually targeted. The cell survival rate of the si-ZNF609+ Anti-miR-153 group was (169.92±13.25)%, higher than (100.00±9.68)% of the si-ZNF609+ Anti-NC group (P<0.001), the ratio of cells in G(0)/G(1) phase and apoptosis rate were (52.01±3.62)% and (8.20±0.87)%, respectively, lower than (64.51±5.17)% and (14.03±1.17)% in the si-ZNF609+ Anti-NC group (P<0.001). The protein expression levels of C-caspase-3 and p21 were (0.42±0.06) and (0.52±0.06), lower than (0.80±0.07) and (0.92±0.10) of the si-ZNF609+ Anti-NC group (P<0.001). The protein expression level of cyclin D1 was (0.68±0.07), higher than (0.39±0.04) in the si-ZNF609+ Anti-NC group (P<0.001). Conclusion: Down-regulation of circZNF609 inhibits the proliferation of diffuse large B-cell lymphoma OCI-LY19 cells and induces apoptosis by targeting miR-153.

Correlation Between Peripheral Blood Biomarkers and Efficacy of PD-1/PD-L1 Inhibitors Treatment on Lung Cancer / 肿瘤防治研究

Objective To investigate the predictive and guiding significance of peripheral blood biomarkers on the therapeutic effects of PD-1/PD-L1 inhibitor treatment on lung cancer patients. Methods We collected the data of 200 lung cancer patients treated with PD-1/PD-L1 inhibitors treatment, including clinical indicators, peripheral blood indicators, efficacy indicators and survival indicators. Results The DCR of patients with non-hepatic metastasis, immune combined chemotherapy, NLR≤2.81 and LDH≤202.5 u/L was higher (P < 0.05). The AUC value of NLR combined with LDH predicting DCR was 0.698 (P < 0.05). Univariate analysis showed that non-hepatic metastasis, first-line immunotherapy, immunotherapy combined with chemotherapy and LDH≤202.5 u/L were related to PFS (P < 0.05). Multivariate analysis showed that the patients with non-hepatic metastasis and LDH≤202.5 u/L had longer PFS (P < 0.05). The significant decrease of NLR and LDH after two cycles of immunotherapy indicated the effectiveness of immunotherapy (P < 0.05). Conclusion NLR≤2.81, LDH≤202.5 u/L, non-hepatic metastasis and immunotherapy combined chemotherapy are positively correlated with immunotherapy efficacy. Non-hepatic metastasis and LDH≤202.5 u/L are independent prognostic factors of the patients treated with immunotherapy. The changes of peripheral blood NLR and LDH are related to the efficacy of PD-1/PD-L1 inhibitors treatment.

Effect of boifilm cleaning agents and muctienzyme cheaning agents in the cleaning of tubular in thstruments / 现代临床护理

Objective To investigate the effect of biofilm cleaning agents and multienzyme cleaning agents in the cleaning of tubular instruments. Methods About 200 pieces of intramedullary nail used in tibial fracture and intramedullary nailing were selected and divided into multienzyme cleaning group and biomembrane cleaning group according to the order of recovery. According to the random number table method, each group had 100 pieces. The biofilm cleaning group was cleaned with biological membrane cleaning agent, and the multienzyme cleaning group was cleaned with multienzyme cleaning agent. The cleaning effect of luminal instruments was observed by means of microscopy, dipstick test and ATP bioluminescence assay. Result The cleaning quality and biofilm removal effect of the biofilm cleaning group were better than those of the multienzyme cleaning group (P<0.05). Conclusion Cleaning the luminal instruments with biomembrane cleaning agent can improve the cleaning quality and prevent the formation of bacterial biofilm.

Experimental study on the interaction between hepatoma cells and hepatic stellate cells / 中国医师杂志

Objective To investigate the effects of the interaction between human hepatoma cells and hepatic stellate cells on their growth state,and study its role of interaction on the progression of hepatocellular carcinoma.Methods Human hepatoma cell line HepG2 and hepatic stellate cell line hepatic stallate cells (HSC)-T6 were used and the methods including methyl thiazolyl tetrazolium (MTT) assay,flow cytometry (FCM) analysis,immunohistochemistry,and electron microscopy were employed in this experiment.The effects of conditioned medium (CM) of HepG2 on the activation and proliferation of HSC were explored.The effects of activated HSC CM on HepG2 proliferation were investigated.The uhrastructural changes of the two co-cultured cells were observed.Results MTT assay result showed that HepG2/HSC CM could promote HSC/HepG2 proliferation.FCM result demonstrated that HepG2/HSC CM could influence the cell cycle distribution in HSC/HepG2.Immunohistochemistry exhibited that after the treatment of HepG2/HSC CM,the expression ofα-smooth muscle actin (α-SMA) in HSC and proliferating cell nuclear antigen (PCNA) in HepG2 were increased.When HepG2 and HSC were co-cultured,the ultrastructure of HSC displayed an activated feature.Conclusions HepG2 cells can induce the activation and proliferation of HSC,and the activated HSC can also stimulate the proliferation of HepG2.Interaction between hepatoma cells and hepatic stellate cells may play an important role in the progression of hepatocellular carcinoma.

Preliminary Clinical Application of a Novel Locking Stylet in Cardiac Lead Extraction / 中国循环杂志

Objective: To explore the safety and efficacy of a novel lead locking device (LLD) in the procedure of cardiac lead extraction for heart rhythm implants. Methods: A total of 6 patients using LLD for cardiac lead extraction in our hospital were retrospectively reviewed. Clinical parameters, the reason of cardiac lead extraction, lead locking stylet condition, outcome of lead extraction and operative complications were summarized. Results: There were 6 patients including 1 female with the median age at 62.5 years. LLD was used and 13 cardiac leads were extracted including 1 scrap electrode wire and 12 functional electrode wire. Among those, LLD was successfully inserted and locked on the top of 11/13 (85%) leads for whole procedure and 2 (15%) leads were not locked for whole procedure; 12 (92 %) leads were completely removed and 1 (8%) lead was partially removed. No severe complications occurred. Conclusion: The novel LLD may safely and effectively extract electrode lead which is beneficial for complete cardiac lead extraction.

The role of PEP-1-SOD1 fusion protein on ischemia-reperfusion injury in isolated perfused rat hearts / 中华心血管病杂志

<p><b>OBJECTIVE</b>The transduction efficiency of the purified PEP-1-SOD1 fusion protein and the effects of PEP-1-SOD1 fusion protein on ischemia reperfusion injury in the isolated perfused rat hearts were investigated.</p><p><b>METHODS</b>The constructed pET15b-SOD1 and pET15b-PEP-1-SOD1 were transformed into BL21 (DE3) for expression and purification of SOD1 and PEP-1-SOD1, respectively. Isolated perfused rat hearts were subjected to 60 min of global ischemia and 30 min of reperfusion and treated with vehicle, 100 micromol/L SOD1 and 25, 50, 100 micromol/L PEP-1-SOD1, respectively. The transduction efficiency was evaluated with immunofluorescent microscopy and Western blot. The enzyme activity of the transduced PEP-1-SOD1 was measured with commercial SOD detection kit. The MDA content in myocardial tissue and the CK activity in coronary exudate at 15 min after reperfusion were also measured. Cardiomyocyte apoptosis was detected with TUNEL. The infarct size was determined in isolated hearts 60 min after reperfusion with TTC staining.</p><p><b>RESULTS</b>Immunofluorescent microscopy and Western blot demonstrated PEP-1-SOD1 was transduced into myocardial tissue in a dose-dependent manner, whereas SOD1 could not be detected in SOD1 group. SOD activity in control, SOD1 group, 25, 50, 100 micromol/L PEP-1-SOD1 groups was (10.06 +/- 0.77) U/mg prot, (10.59 +/- 0.71) U/mg prot, (32.29 +/- 1.42) U/mg prot, (43.16 +/- 1.16) U/mg prot, (55.14 +/- 1.59) U/mg prot, respectively. MDA content in corresponding groups was (1.48 +/- 0.19) nmol/mg prot, (1.39 +/- 0.11) nmol/mg prot, (1.01 +/- 0.14) nmol/mg prot, (0.73 +/- 0.13) nmol/mg prot, (0.50 +/- 0.06) nmol/mg prot, respectively. CK activity in corresponding groups was (1.73 +/- 0.58) U/mg prot,(1.68 +/- 0.14) U/mg prot,(1.40 +/- 0.28) U/mg prot,(0.97 +/- 0.39) U/mg prot, (0.61 +/- 0.56) U/mg prot, respectively. Cardiomyocyte apoptotic index in corresponding groups was (17.25 +/- 0.75)%, (16.63 +/- 1.07)%, (11.50 +/- 0.57) U/mg prot, (6.50 +/- 0.63) U/mg prot, (4.13 +/- 0.52)%, repectively. The percentage of myocardial infarction area was (55.13 +/- 2.18)%, (52.13 +/- 2.59)%, (33.88 +/- 2.06)%, (25.50 +/- 2.16)%, (15.38 +/- 1.14)%, respectively. Compared with control group and SOD1 group, all P < 0.01 These results demonstrated the enzyme activity of the transduced PEP-1-SOD1 was significantly increased in a dose-dependent manner and the MDA content, CK activity, the cardiomyocyte apoptotic index and the infarct size was decreased siginificantly in PEP-1-SOD1 pretreatment groups compared with SOD1 group.</p><p><b>CONCLUSION</b>The native, biologically active form of PEP-SOD1 fusion protein could be effectively transduced into the isolated rat hearts subjecting ischemia reperfusion injury in a dose-dependent manner. The transduced PEP-1-SOD1 has protective effects on ischemia reperfusion injury in the isolated rat hearts.</p>

Effect of Different Proportions of Mixed Blood Exchange Transfusion on Blood Internal Environment in Neonates with Hemolytic Disease / 实用儿科临床杂志

Objective To explore the effect of different proportions of mixed blood exchange transfusion on blood circulation in neonates with hemolytic disease.Methods Thirty-one newborn infants with hemolytic disease were treated by peripheral arteriovenous synchronization of exchange transfusion with different proportions mixed blood.AB type plasma was mixed with O type red blood cell(RBC) washing.The proportion for the treatment group was 1:1(the O type RBCs 2 U:the AB type plasma 200 mL),by exchange transfusion of haplotypes,in accordance with 80?mL/kg;the proportion for control group was 2:1(the O type RBC 4 U:the AB type plasma 200 mL),by exchange transfusion of double in accordance with 150-180 mL/kg.The indicators were detected,such as the exchange rate of neonatal serum bilirubin,RBC,hemoglobin(Hb),hematocrit(HCT),and the exchange transfusion quantity and days of hospitalization before and after the exchange transfusion were analyzed.Results The exchange rate of serum bilirubin of treatment group and control group was (44.92?3.99)% and (45.69?5.06)%,respectively,there was no significant difference between 2 groups(P=0.639),there was no significant difference of hospitalization days[(8.13?1.13) d vs(8.19?0.91) d]between 2 groups(P=0.884).After exchange transfusion in treatment group,the average level of the RBC,Hb and HCT were increased(P

Store-operated Ca2+ channels in rat colonic smooth muscle cells / 中国应用生理学杂志

<p><b>AIM</b>To study whether store-operated Ca2+ channel (SOC) is present in rat colonic smooth muscle cells.</p><p><b>METHODS</b>Intracellular Ca2+ ([Ca2+]i) changes induced by thapsigargin- or caffeine-activated SOC channel were measured in enzymatically dissociated rat colonic smooth muscle cells with the fluorescent indicator Fura-2/AM.</p><p><b>RESULTS</b>In the absence of external Ca2+ , the sarco-endoplasmic reticulum Ca2+ pump inhibitor thapsigargin (1 micromol/L) and ryanodine receptor (RyR) activator caffeine both transiently elevated [Ca2+]i from (68.32 +/- 3.43) nmol/L to (240.85 +/- 12.65 ) nmol/L, (481.25 +/- 34.77) nmol/L. A subsequent reintroduction of Ca2+ into the extracellular solution resulted in [Ca2+]i further elevated to (457.55 +/- 19.80) nmol/L, (1005.93 +/- 54.62) nmol/L; (643.88 +/- 34.65) nmol/L, (920.16 +/- 43.25) nmol/L, respectively. And the elevated response was blocked by lanthanum (1 mmol/L), but was insensitive to L-type voltage calcium channels blocker verapamil and membrane depolarization.</p><p><b>CONCLUSION</b>SOC activated by store depletion are present in rat colonic smooth muscle cells. And we further prove the existence of such Ca2+ channels in excitable cells.</p>

Capacitative Ca²⁺ entry is involved in ACh-induced distal colon smooth muscle contraction in rats / 生理学报

Contraction of smooth muscle cells is triggered by an increase in cytosolic Ca(2+) upon agonist stimulation. Ca(2+) influx across the plasma membrane constitutes a major component of the agonist-induced response in smooth muscle cells. Traditionally, voltage-operated Ca(2+) channel (VOCC) is considered as the channel mediating the Ca(2+) entry. However, this view has been challenged by recent discoveries, which demonstrated that other types of ion channels, such as store-operated and/or receptor-operated Ca(2+) channels (SOCC and/or ROCC), also participate in Ca(2+) response induced by agonists in smooth muscle cells. SOCC is defined as the channel activated in response to the depletion of the internal Ca(2+) stores, an event secondary to G protein coupled receptor or receptor tyrosine kinase stimulation. The Ca(2+) flow mediated by SOCC is termed as capacitative Ca(2+) entry (CCE). Previous study from other group has demonstrated that VOCC played a predominant role in ACh-induced contraction of distal colon smooth muscle in guinea pig. However, whether SOCC participates in the agonist-induced contractile response in this particular tissue is unknown. The present study was performed to investigate the role of CCE in ACh-induced mechanical activity of distal colon smooth muscle in rats. The contractile function of the smooth muscle was assessed by measuring isometric force of isolated rat distal colon rings. We showed that both high extracellular K(+) (40 mmol/L) and ACh (5 mumol/L) evoked striking contraction of the smooth muscle. The contractile responses were almost abolished by removal of extracellular Ca(2+) with ethylene glycol-bis(2-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA), suggesting a critical contribution of extracellular source of Ca(2+) to the contraction. Verapamil (5 mumol/L), an L-type VOCC blocker, significantly attenuated, but didn't completely eliminate the high K(+)- and ACh-induced contraction (74% and 41% for high K(+) and ACh, respectively), indicating that additional channels might be involved in the contractile mechanism. Furthermore, ACh only induced transient contractions in the absence of extracellular Ca(2+). Readmission of Ca(2+) into the extracellular compartment resulted in a significant and sustained increase in the tension of the smooth muscle. This response was not affected by verapamil (5 mumol/L) and Cd(2+) (5 mumol/L), both of which efficiently block VOCC at the doses. However, La(3+), a known inhibitor of SOCC, significantly suppressed the Ca(2+) readdition-induced contraction in a dose-dependent manner. On the basis of these results, we conclude that contraction of smooth muscle in the distal colon is regulated by multiple Ca(2+) channels. In addition to VOCC-mediated Ca(2+) influx, SOCC-mediated CCE participates in agonist-induced contractile response of distal colon smooth muscle in rats.

Bombesin-mediated non-cholinergic late slow excitatory postsynaptic potentials in guinea pig inferior mesenteric ganglion in vitro / 生理学报

The effect of bombesin (BOM) on non-cholinergic excitatory synaptic transmission of the guinea pig inferior mesenteric ganglion (IMG) was investigated by intracellular recording. Repetitive stimulation of the colon nerves (1 ms, 25 Hz, 4 s) elicited a burst of action potentials, which was followed by a long-lasting depolarization in 74.3% (52/70) of the IMG neurons. The depolarization was not blocked by nicotinic (d-tubocurarine, 100 micromol/L) and muscarinic (atropine, 1 micromol/L) antagonists, but was eliminated in a low Ca(2+)/high Mg(2+) Krebs solution, indicating that the depolarization was due to the release of non-cholinergic transmitters. Superfusing the ganglia with BOM (10 micromol/L, 1 min) induced a slow depolarization in 66.5% (109/164) neurons tested. The BOM response was not appreciably changed in low Ca(2+)/high Mg(2+) Krebs solution (n=6, P>0.05), suggesting that BOM depolarized the neurons by acting directly on the postsynaptic membrane rather than via a release of other endogenous depolarizing substances. In a total of 102 cells that exhibited late slow excitatory postsynaptic potential (ls-EPSP), superfusion of the ganglia with BOM produced a membrane depolarization in 82 neurons (80%), while the remaining 20 cells (20%) exhibited no response to BOM. In 18 neurons with ls-EPSP, 4 (22%) neurons were sensitive to both BOM and SP; 6 (33%) and 5 (28%) neurons were only sensitive to BOM and SP, respectively. The remaining 3 (17%) neurons were insensitive to both BOM and SP. Membrane resistance (Rm) had no apparent change in 47.3%, 59.5 % of the neurons tested during the ls-EPSP (n=55) and BOM depolarization (n=84), respectively, but had a marked decrease in 38.2%, 27.4%, and a marked increase in the remaining 14.5%, 13.1% of the neurons. However, when the Rm change accompanying ls-EPSP was compared with that accompanying BOM depolarization (n=20) in the same neuron, the changes in Rm were always parallel. Moreover, ls-EPSP (n=6) and BOM depolarization (n=8) were all augmented by conditioning hyperpolarization. The extrapolated values of the reversal potentials of ls-EPSP and BOM depolarization were 46.0+/-8.0 and 50.0+/-7.0 mV (n=8, P>0.05), respectively. In 14 BOM-sensitive neurons, a ls-EPSP was elicited by repetitive colon nerve stimulation. Superfusion of BOM (10 micromol/L) in these cells initially caused a large depolarization and then membrane potential gradually subsided to resting level in the continuous presence of BOM. Stimulation of the presynaptic nerves at this time failed to elicit a detecable ls-EPSP in 2 neurons and induced a much smaller one in 10 cells, while the ls-EPSP in the remaining 2 neurons was not appreciably affected. On the other hand, prolonged superfusion of BOM had no effect on the amplitude and duration of ls-EPSP in 6 BOM-insensititive neurons studied (P>0.05). The amplitude and duration of SP-induced depolarization were not altered by prolonged superfusion of BOM (n=4, P>0.05) Superfusion of tyr(4) D-phe(12) bombesin (1 micromol/L, 10 15 min), a BOM receptor antagonist, did not cause any noticeable changes in passive membrane properties nor block nicotinic f-EPSPs, but markedly suppressed (n=5) or completely abolished (n=11) BOM depolarization in all 16 neurons tested Similarly, tyr(4) D-phe(12) bombesin partially or completely antagonized the ls-EPSP in 9 out of a total of BOM sensitive neurons (n=11). The ls-EPSP elicited in the remaining two neurons was insignificantly affected by this drug. However, following 10 20 min of wash with Krebs solution the ls-EPSP was reversed. In contrast, superfusion of the ganglia with tyr(4) D-phe(12) bombesin did not change the amplitude and duration (P>0.05) of ls-EPSP in 10 BOM-insensitive cells. Similarly, the amplitude and duration of SP-induced depolarization were not appreciably affected by tyr(4) D-phe(12) bombesin (n=6, P>0.05). In conclusion, our results indicate that BOM may be another transmitter mediating the ls-EPSP in the guinea pig IMG and that there is no cross-desensitization of BOM receptors and SP receptors.

Effect of alpha-difluoromethylornithine on growth characteristics and expression of ALT-04ag gene of human lung carcinoma cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effect of polyamine biosynthesis inhibition on growth characteristics of human lung carcinoma cells and its correlation with the expression of human lung carcinoma associated antigen ALT-04ag gene.</p><p><b>METHODS</b>The gene expression was detected by RT-PCR and immunocytochemical tests. The cell growth characteristics were studied by cell growth curves, morphological observation, FCM analysis and DNA electrophoresis.</p><p><b>RESULTS</b>Human lung squamous carcinoma cells L78 treated with 5 mmol/L alpha-difluoromethylornithine (DFMO) for 5 days showed significant growth inhibition and apoptosis induction. The mRNA and protein expressions of ALT-04ag gene in the cells were downregulated, while these changes resulted from DFMO treatment were prevented by provision of DFMO along with exogenous putrescine.</p><p><b>CONCLUSION</b>The effect of polyamine biosynthesis inhibition induced by DFMO restrains the growth characteristics and promotes apoptosis of human lung carcinoma L78 cells, which is associated with down regulation of ALT-04ag gene expression.</p>