Comparison of values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus testing in screening of high-grade cervical lesions / 肿瘤研究与临床

Objective:To investigate the screening values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus (HR-HPV) testing for high-grade cervical lesions. Methods:The clinical data of 622 patients who underwent cervical thin-layer liquid-based cytology (TCT) and HR-HPV testing in General Hospital of Taiyuan Iron and Steel (Group) Co., Ltd. from March 2019 to July 2021 were retrospectively analyzed. The remaining cytological specimens were detected by P16/Ki-67 double staining and P16 INK4α single staining. Among them, 334 patients with TCT results suggesting atypical squamous cell of undetermined significance (ASCUS) and above and HPV-positive underwent colposcopy pathological biopsy. Using pathological results as reference, the positive predictive value, sensitivity, specificity and accuracy of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of high-grade squamous intraepithelial neoplasia (HSIL) and cervical cancer were compared. Results:Taking the results of histopathology as references, combined with the results of TCT, 31 of 622 patients were HSIL, of which 22 (71.0%) were positive for P16/Ki-67 double staining, 23 (74.2%) were positive for P16 INK4α single staining, and 25 (80.6%) were positive for HR-HPV testing; 4 cases were cervical cancer, and the positive rates of the three detection methods were all 100.0% (4/4). Among 622 patients, the positive rates of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of HSIL and cervical cancer were 13.99% (87/622), 25.40% (158/622) and 21.38% (133/622); the positive predictive values were 29.89%, 17.09% and 21.08%; the accuracies were 91.19%, 78.94% and 83.28%; the specificities were 89.77%, 77.98% and 82.46%; the sensitivities were 74.29%, 77.14% and 82.86%. The positive rate, positive predictive value, specificity and accuracy of P16/Ki-67 double staining were higher than those of P16 INK4α single staining and HR-HPV testing, and the differences were statistically significant ( z values were -5.062 and -3.418, 2.328 and 2.450, 5.436 and 3.570, 6.043 and 4.161, all P < 0.05); the sensitivity of HR-HPV testing was higher than that of P16/Ki-67 double staining and P16 INK4α single staining, but the differences were not statistically significant ( z values were -0.890 and 1.017, both P > 0.05). Conclusions:HR-HPV testing is more suitable for primary cervical lesion screening; P16/Ki-67 double staining can be used as a potential combined cell screening tool or an effective triage tool; P16 INK4α single staining has certain limitations.

Study on the effect of costunolide on sensitivity of chronic myeloid leukemia K562/ADR cells to doxorubicin via p38-MAPK pathway / 白血病·淋巴瘤

Objective:To explore the effect of costunolide on sensitivity of chronic myeloid leukemia cell line K562/ADR to doxorubicin and its mechanism.Methods:K562/ADR cells in the logarithmic phase were used, and the cells were treated with different concentrations of costunolide, doxorubicin or costunolide combined with doxorubicin for 72 h. The cell proliferation was detected by CCK-8 method, the cell proliferation rate was calculated, and the half inhibitory concentration (IC 50) of the two drugs was obtained. The cells were treated with 10 μmol/L costunolide, 10 μmol/L doxorubicin or costunolide combined with doxorubicin for 48 h, the apoptotic rate was detected by flow cytometry, and the expression level of p38-MAPK pathway related proteins was detected by Western blot. Results:The cell proliferation rate in the costunolide combined with doxorubicin group was lower than that in the corresponding concentration of the two drugs alone groups, and the differences were statistically significant both ( P < 0.05). The IC 50 of doxorubicin for K562/ADR cells was (13.50±0.86) μmol/L, and costunolide was (7.30±0.55) μmol/L ( t = 7.044, P = 0.002). The results of flow cytometry showed that the apoptosis rate of K562/ADR cells in the 10 μmol/L costunolide combined with 10 μmol/L doxorubicin group was higher than that of the blank control group, costunolide alone group and doxorubicin alone group, and the differences were statistically significant [(19.68±3.21)% vs. (2.96±0.87)%, (9.34±2.89)%, (9.18±2.13)%, all P<0.01]. Compared with the 10 μmol/L costunolide alone group and the 10 μmol/L doxorubicin alone group, the expression of the apoptosis inhibitor protein bcl-2 in the two-drug combination group was down-regulated, and the expressions of bad, p-p38, cleaved-caspase-3 and cleaved-PARP proteins were up-regulated. Conclusion:Costunolide can enhance the inhibitory effect of doxorubicin on the proliferation of K562/ADR cells and promote doxorubicin-induced apoptosis, which may reverse the drug resistance of K562/ADR cells by regulating the p38-MAPK pathway.

Preventive effect of anticoagulant therapy and catheter ablation on dementia in patients with atrial fibrillation / 国际脑血管病杂志

Atrial fibrillation (AF) is the most common type of arrhythmia, and its prevalence increases with age. Studies have shown that AF is associated with an increased risk of dementia. There is increasing evidence of potential treatment strategies to prevent dementia in patients with AF. This article reviews the preventive effects of anticoagulant therapy and catheter ablation on dementia in patients with AF.

Autophagy activation attenuates the neurotoxicity of local anesthetics by decreasing caspase-3 activity in rats / A ativação autofágica atenua a neurotoxicidade dos anestésicos locais ao diminuir a atividade da caspase‐ 3 em ratos

Abstract Background and objectives The mechanisms by which local anesthetics cause neurotoxicity are very complicated. Apoptosis and autophagy are highly coordinated mechanisms that maintain cellular homeostasis against stress. Studies have shown that autophagy activation serves as a protective mechanism in vitro. However, whether it also plays the same role in vivo is unclear. The aim of this study was to explore the role of autophagy in local anesthetic-induced neurotoxicity and to elucidate the mechanism of neurotoxicity in an intrathecally injected rat model. Methods Eighteen healthy adult male Sprague-Dawley rats were randomly divided into three groups. Before receiving an intrathecal injection of 1% bupivacaine, each rat received an intraperitoneal injection of vehicle or rapamycin (1 mg.kg-1) once a day for 3 days. The pathological changes were examined by Haematoxylin and Eosin (HE) staining. Apoptosis was analysed by TdT-mediated dUTP Nick-End Labelling (TUNEL) staining. Caspase-3, Beclin1 and LC3 expression was examined by Immunohistochemical (IHC) staining. Beclin1 and LC3 expression and the LC3-II/LC3-I ratio were detected by western blot analysis. Results After bupivacaine was injected intrathecally, pathological damage occurred in spinal cord neurons, and the levels of apoptosis and caspase-3 increased. Enhancement of autophagy with rapamycin markedly alleviated the pathological changes and decreased the levels of apoptosis and caspase-3 while increasing the expression of LC3 and Beclin1 and the ratio of LC3-II to LC3-I. Conclusions Enhancement of autophagy decreases caspase-3-dependent apoptosis and improves neuronal survivalin vivo. Activation of autophagy may be a potential therapeutic strategy for local anaesthetic-induced neurotoxicity.

Resumo Introdução e objetivos Os mecanismos de neurotoxicidade dos anestésicos locais são complexos. A apoptose e a autofagia são mecanismos altamente organizados que mantêm a homeostase celular durante o estresse. Estudos revelam que a ativação da autofagia atua como mecanismo de proteção in vitro. Não está claro se a autofagia também desempenha essa função in vivo. O objetivo deste estudo foi analisar o papel da autofagia na neurotoxicidade induzida por anestésico local e esclarecer o mecanismo dessa neurotoxicidade utilizando um modelo de injeção intratecal em ratos. Métodos Dezoito ratos Sprague‐Dawley machos adultos saudáveis foram divididos aleatoriamente em três grupos. Antes de receber a injeção intratecal de bupivacaína a 1%, cada rato recebeu injeção intraperitoneal de veículo ou rapamicina (1 mg.kg‐1) uma vez ao dia durante 3 dias. As alterações patológicas foram examinadas por coloração com Hematoxilina e Eosina (HE). A apoptose foi analisada por coloração com o método dUTP Nick‐End Labeling (TUNEL) mediado por TdT. A expressão de caspase‐3, Beclin1 e LC3 foram examinadas por coloração Imunohistoquímica (IHQ). A expressão de Beclin1 e LC3 e a razão LC3‐II/LC3‐I foram detectadas por análise de western blot. Resultados Após a injeção intratecal de bupivacaína, ocorreu lesão patológica nos neurônios da medula espinhal e os níveis de apoptose e caspase‐3 aumentaram. A ativação da autofagia causada pela rapamicina mitigou de forma expressiva as alterações patológicas e diminuiu os níveis de apoptose e caspase‐3, aumentando a expressão de LC3 e Beclin1 e a razão LC3‐II/LC3‐I. Conclusões O aumento da autofagia diminui a apoptose dependente da caspase‐3 e melhora a sobrevivência neuronal in vivo. A ativação da autofagia pode ser uma estratégia terapêutica potencial para a neurotoxicidade induzida por anestésicos locais.

Effect of asiaticoside on hyperoxia-induced bronchopulmonary dysplasia in neonatal rats and related mechanism / 中国当代儿科杂志

OBJECTIVE@#To study the protective effect of asiaticoside against hyperoxia-induced bronchopulmonary dysplasia in neonatal rats based on the microRNA-155 (miR-155)/suppressor of cytokine signaling-1 (SOCS1) axis.@*METHODS@#Neonatal rats were randomly divided into a control group, a model group, a low-dose asiaticoside group (10 mg/kg), a middle-dose asiaticoside group (25 mg/kg), a high-dose asiaticoside group (50 mg/kg), and a budesonide group (1.5 mg/kg), with 12 rats in each group. All rats except those in the control group were exposed to a high concentration of oxygen for 14 days to establish a neonatal rat model of bronchopulmonary dysplasia. The low-, middle-, and high-dose asiaticoside groups were given asiaticoside at different doses by gavage, and those in the budesonide group were given budesonide aerosol treatment. Hematoxylin and eosin staining was used to observe lung tissue development and measure radial alveolar count (RAC) and mean linear intercept (MLI). Superoxide dismutase (SOD) and malondialdehyde (MDA) detection kits were used to measure the levels of SOD and MDA in lung tissue. ELISA was used to measure the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Quantitative real-time PCR was used to measure the mRNA expression of miR-155 and SOCS1 in lung tissue. Western blotting was used to measure the protein expression of SOCS1 in lung tissue.@*RESULTS@#Compared with the control group, the model group had the symptoms of bronchopulmonary dysplasia such as a disordered structure of lung tissue, enlargement of alveolar fusion, uneven alveolar septa, enlargement of average alveolar space, and a reduction in alveolar number. The model group also had significant increases in MLI, MDA level in lung tissue, serum levels of IL-6 and TNF-α, and miR-155 level in lung tissue (P0.05).@*CONCLUSIONS@#Asiaticoside can alleviate inflammation injury induced by hyperoxia in neonatal rats and improve the symptoms of bronchopulmonary dysplasia in a dose-dependent manner, possibly by down-regulating the expression of miR-155 and up-regulating the expression of SOCS1.

Biopharmaceuticals classification system (BCS) of periplocin, periplocymarin and periplogenin / 中草药

Objective To investigate the biopharmaceutical classification of periplocin, periplocymarin and periplogenin by biopharmaceuticals classification system (BCS). Methods The solubility and permeability of periplocin, periplocymarin and periplogenin were studied by using experimental and computer prediction software Pipeline Pilot 8.5 and ChemDraw. These three substances were sorted based on experimental and predicted values by BCS classification method. Results According to the test results, periplocin, periplocymarin and periplogenin were classified as BCS III, which are different from the classification results predicted by different software. The permeability results based on ClgP was consistent with the experimental results; The solubility based on lgCs and the permeability based on AlgP and lgD were opposite to the experimental results. Conclusion The periplocin, periplocymarin and periplogenin are BCS III drugs, the solubility is decreased in turn, but they are still highly soluble components. The permeability is the key factor affecting its absorption. The predicted value of biopharmaceutical properties of active ingredients of A-type cardiac glycosides based on the chemical structure are significant different with the test results, for the BCS study and the in vitro and in vivo correlation evaluation of oral absorption of traditional Chinese medicine preparations containing such ingredients, it is recommended to use a variety of methods to correct the data and increase the reliability of the results.

Mechanism of angiotensin II (Ang II) on the proliferation of human hepatoma cell line HepG2 cells / 中华肝脏病杂志

Objective@#To study the effect and mechanism of angiotensin (Ang II) on the proliferation of human hepatocellular carcinoma HepG2 cells.@*Methods@#The effects of different concentrations of Ang II's (10-8-10-4 mol/L) on proliferated hepatocellular carcinoma HepG2 cells were detected by CCK-8 assay. The expression of angiotensin II type 1 receptor (AT1) protein and activation of ERK1/2 protein in hepatocellular carcinoma HepG2 cells after processing with Ang II were assayed by Western blot. The cells were pretreated with candesartan (AT1 receptor antagonist), sorafenib (Raf kinase inhibitor) and PD98059 (ERK1/2 inhibitor) for 1.5 h and then Ang II (10-6 mol/L) was added. CCK-8 assay was used to determine whether it could reverse the proliferation of Ang II, and ERK phosphorylation levels were detected by Western blot. The changes in Bcl-2 and c-myc gene expression before and after Ang II processing were detected by Rt-PCR. According to different data, t-test, one-way analysis of variance or SNK method were used for statistical analysis.@*Results@#HepG2 cells treated with different concentrations of Ang II promoted cell proliferation after 24h and 48h. After 24 h, cell vitality was strongest with Ang II concentration 10-5 mol/L and the absorbance value was 0.990 8±0.097 8; and again after 48 h, the cell viability was strongest with Ang II concentration 10-6 mol/L and the absorbance value was 1.302 7 ± 0.030 9. Moreover, the pro-proliferation effect of Ang II on HepG2 cells blocked candesartan, sorafenib and ERK1/2 isolated inhibitors. After treatment with 10-6 mol/L Ang II, Western blot showed that Ang II significantly promoted AT1 receptor expression and phosphorylation of ERK1/2 protein confirmed that Ang II activated the AT1/RAF/ERK1/2 signaling pathway. In addition, Rt-PCR detection showed that the downstream of Bcl-2 and c-myc genes expressions rose significantly when the concentration of Ang II ranged from 10-8 to 10-6 mol/L.@*Conclusion@#Ang II can promote the proliferation of HepG2 cells by activating AT1/Raf /ERK1/2 signaling pathway and enhance the downstream of Bcl-2 and c-myc gene expression.

Inhibitory effect of favipiravir on canine distemper virus replication in vitro / 军事医学

Objective To investigate the inhibitory effect of favipiravir (T-705) on canine distemper virus (CDV) replication in Vero cells and DH82 cells.Methods The growth curves of CDV-11 strains from canine and CDV-3 strains from mink in Vero cells and DH82 cells were determined with indirect immunofluorescence assay and 50％ endpoint titration.The viability of Vero and DH82 cells was determined using the Cell Counting Kit-8.CDV inhibition at different concentrations of T-705 at different time points was measured .Results Cytotoxicity data showed that there was a moderate decline of viability in Vero cells after T-705 treatment, but no apparent cytotoxicity in DH 82 cells.T-705 significantly inhibited the replication of CDV-3 and CDV-11 in both Vero cells and DH82 cells in the test range of 2.441-1250 μg/ml. T-705 exhibited effective and stable antiviral activity when given at different time points post virus challenge .Conclusion Our results demonstrate that T-705 has effective antiviral activity and may be a promising anti-CDV drug candidate .

Effect of duck Tembusu virus infection on exosomes of BHK-21 cells / 中国免疫学杂志

Objective:To explore the effect of duck Tembusu virus infection on secretion of exosomes in BHK-21 cells and the pathogenesis of the Tembusu virus.Methods:The exosomes were collected and purified from the culture supernatant of BHK-21 cells infected with duck Tembusu virus AH-F10 strain and the control BHK-21 cells by PEG precipitation method respectively.The purified exosomes were identified by electron microscopy,Western blot assay and mass spectrometry.Results: The classical exosome particle morphology was observed with hyperchromic cup-shaped vesicles and average particle size of 30-160 nm in diameter under transmission electron microscopy.The mean size of the exosome from the infected cells were bigger than the mean size of the exosome from the unin-fected.Western blot assay demonstrated that CD9 and CD63 were detected in purified exosomes as exosome marker molecula.A total of 106 proteins were identified by mass spectrometry assay,84 proteins of infected BHK-21 cells exosome,49 proteins of the uninfected, and the infected and the uninfected BHK-21 share 27 common proteins on exosomes.Conclusion:Duck Tembusu virus infection affect the exosome secretion of cells in connection to the particle size and protein molecular composition.This experiment can lay the foundation for further research of Tembusu virus infection and pathogenesis.

Experimental observation of human adipose mesenchymal stem cells transplantation in the treatment of acute necrotic pancreatitis in rats / 中华胰腺病杂志

Objective To observe the effect of human adipose mesenchymal stem cells (hADMSCs) on pancreatic tissue repair and inflammatory reaction of acute necrotic pancreatitis (ANP) in rat, and explore the possible mechanism.Methods Isolation and purification of hADMSCs and flow cytometry to detect the the surface markers including CD90, CD29, CD34 and CD45 were performed.Eighty SD male rats with the body weight of 170~210 g were randomly divided into 4 groups.There were 8 rats in the control group, 24 rats in other group.Control group underwent no treatment;sham operation group underwent intestinal wall stirring and then abdominal closure;ANP model group was established by open abdominal retrograde injection of sodium taurocholate into bile duct;and in hADMSCs group, DAPI labeled hADMSCs were injected by tail vein into the rat at 12 h after sodium taurocholate injection.The survival of the rats, and gross morphological and pathological changes of the pancreas was observed at 12, 24, and 48 h, and the serum TNF-α, IL-6, IL-10 and amylase were detected.The distribution of hADMSCs in the pancreas, liver and lung was examined in hADMSCs group.Results Rats in control group and sham operation group were all alive.In ANP group, 5 and 11 rats were dead at 24 and 48 h, respectively, and in hADMSCs group 12 rats were dead at 48 h.Compared with ANP group, the difference was not statistically significant (P>0.05).The pathological changes of the pancreas were significantly less severe in hADMSCs group than in ANP group.In hADMSCs group, the amylase at 12, 24 and 48 h was(999±110 )、(1 831±110)、(3 991±130 )U/L;TNF-α level was (62.40±2.35), (80.51±4.51) and (93.46±6.60)ng /L;IL-6 was (60.46±7.34), (80.61±8.40) and(100.58±9.49)ng /L;and these were all significantly lower than those in ANP model group [amylase (2 402±146), (3 292±137) and (5 632±112)U/L;TNF-α(87.13±3.39), (105.41±10.06), (114.57±3.06)ng/L;IL-6 (70.67±10.90)、(107.61±10.53)、(145.34±10.48)U/L], and the differences were all statistically significant (all P<0.05).IL-10 in hADMSCs group was (56.63±6.35), (81.32±5.96), (100.26±6.51)ng/L, which were increased compared with those in ANP model group [(45.26±8.04), (68.25±8.42), (80.38±5.71)ng/L], and the difference was statistically significant (all P<0.05).hADMSCs can migrate to the pancreas, liver, lungs and other damaged tissue, with most in pancreatic tissue, less in lung tissue, and least in liver tissue.Conclusions The mechanism of hADMSCs in repairing pancreatic tissue injury was associated with inhibiting TNF-α and IL-6 secreting and increasing IL-10, thus reducing inflammatory reaction.

Interferon-γ and interleukin-4 detected in serum and saliva from patients with oral lichen planus / 国际口腔科学杂志·英文版

Our previous salivary study had demonstrated an apparent T helper 2 (Th2)-predominance in saliva of oral lichen planus (OLP) patients and suggested a potential of salivary interleukin-4 (IL-4) as a biomarker for monitoring disease severity. To further determine the consistency of Th1/Th2 bias of OLP, this study investigated the expression profile of interferon-γ (IFN-γ) and IL-4 in serum and the relationship of the serum levels of these cytokines with their saliva partners. Sixty ethnic Chinese patients with OLP (40 of the erythematous/ulcerative form and 20 of the reticular form) were recruited for this study, with 40 age-sex-matched healthy volunteers as control group. IFN-γ and IL-4 levels in serum and paired saliva samples were screened by enzyme-linked immunosorbent assay. OLP patient showed a low-level IFN-γ but high-level IL-4 expression profile in both serum and saliva, with a lower IFN-γ/IL-4 ratio. Serum IL-4 level in the erythematous/ulcerative group was significantly higher than that in the reticular group. Serum levels of IFN-γ and IL-4 were significantly and positively correlated with their saliva partners. These results provided more evidence for Th2 cytokine-predominant immune imbalance in OLP, as well as the potential of IL-4 as the biomarker for monitoring severity of OLP.

A clinical study on the right inferior hepatic vein / 中华肝胆外科杂志

Objective To observe the anatomy of the inferior right hepatic veins (IRHV).Methods The IRHVs were divided into 3 groups according to the location where they entered into the retrohepatic inferior vena cava at: the upper 1/3, middle or lower 1/3. The incidence, number, caliber, extrahepatic length and the relationship between the major hepatic veins (the right, middle and the left hepatic veins) and the IRHV were observed and measured in 60 adult cadavers. Results The incidence of IRHV was 83.33% with an average diameter of 2.62-18.46(14.32±1.21)mm. Its extrahepatic length was 3.26-47.65 (10.78±7.81)mm. There was a marked negative correlation between the diameter of the IRHV and its number, a marked negative correlation between the diameter of the IRHV and the diameter of the right hepatic vein and a marked positive correlation between the number of the IRHV and the diameter of the right hepatic vein. Conclusions There were high variations in the incidence and anatomy of the IRHV which were related to the diameter of the right hepatic vein. The IRHV was not to be torn during liver resection and should be reconstructed in right liver grafts.

Two Simulation Studies on the Closed and Open Peritoneum in Abdomenal Operations / 医学研究杂志

Objective To discuss two simulation studies on the closed and open peritoneum in abdomenal operations. Meth-ods We selected 120 domestic female rabbits from animal laboratory of Qi Qi Ha’er Medical College at random and divided them into four groups(according to whether the peritoneum was open or not,the degree of peritoneum defect at the right side of incision and the existence of peritoneum hemorrhagic focus ),with 30 cases in each group. Group Ⅰ: no peritoneum suture and making a defect of 4cm?3cm at the right side of the incision; Group Ⅱ: no peritoneum suture and making a defect of 4cm?3cm at the right side of the incision,with a hemorrhagic focus at peritoneum defect;Group Ⅲ: with peritoneum suture; Group Ⅳ:with dense and compact peritoneum suture.And then we analyzed postoperative peritoneum healing progress. Results Observeations of the incision infection by the naked eye were that one case was identified in group Ⅰ,Ⅲ and Ⅳ;and a significant difference of occurrence rate was identified between the fallowing groups: groupⅠandⅡ(P

Surgical treatment of Mirizzi syndrome (a report of 43 cases) / 中国普通外科杂志

Objective To study the pathologic feature and rational diagnosis and treatment of Mirizzi Syndrome. Method The clinical data of 43 cases treated by surgery were retrospectively analysed. Results All the 43 cases underwent operation, including partial cholecystecomy in 8 cases, cholecystectomy in 16cases , cholecystectomy plus common bile duct exploration with T tube drainage in 9 cases, choledochojejunostomy in 10 cases. Of the 43 cases, 36 cases were followed up for 1～5 years. Of them, 29 cases were in excellent, 6 cases in good and 1 case in poor. Conclusions The pathologic type of Mirizzi Syndrome is variant. It is difficult to make a definite diagnoses before operation. So vary imaginal technique should be adopted. Different operative procedures should be used according to patients' pathologic type.

Expression of VEGF mRNA induced by angiotensinⅡ in human hepatic cancer cell line / 中国普通外科杂志

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of vascular endothelial growth factor (VEGF) in human hepatic cancer cell line Hep G2.Methods Cultured Hep G2 cells were treated by AngⅡ with various concentrations and were collected at different time points.Then total RNA was extracted.The expression of VEGF mRNA in cultured Hep G2 was determined by relative quantitative reverse transcription polymerase chain reaction(RT-PCR),and the proliferation was detected by methyl thiazolyl tetrazolium (MTT).Results AngⅡ stimulated the proliferation of HepG2 cells,and enhanced the expression of VEGF mRNA (P