Objective:To discuss the role of M2 macrophages in epithelial-mesenchymal transition(EMT)and cisplatin(DDP)resistance in the non-small cell lung cancer(NSCLC),and to clarify the regulatory mechanism of nuclear factor κB(NF-κB)signaling pathway.Methods:The human monocytic leukemia THP-1 cells were selected and differentiated into M0 macrophages by phorbol myristate acetate(PMA)induction,followed by M2 macrophage polarization through interleukin(IL)-4 and IL-13 stimulation.Western blotting and immunofluorescence methods were used to detect the protein expression levels of CD163,CD86,and arginase-1(Arg-1)in M0 and M2 macrophages.The human NSCLC A549 cells were co-cultured non-contactly with M0 or M2 macrophages in Transwell chambers,and the cells were divided into A549+M0 group(A549 cells co-cultured with M0 macrophages),A549+M2 group(A549 cells co-cultured with M2 macrophages),and A549+M2+BAY11-7082 group(A549 cells co-cultured with M2 macrophages and treated with 10 mmol·L-1 NF-κB inhibitor BAY11-7082).Wound healing assay was used to detect the wound healing rate of the A549 cells in various groups;Transwell assay was used to detect the number of invasion A549 cells in various groups;cell counting kit-8(CCK-8)assay was used to detect the inhibitory rate of proliferation and half maximal inhibitory concentration(IC50)value of the A549 cells after treated with DDP in the co-culture system;Western blotting method was used to detect the expression levels of vimentin,E-cadherin,N-cadherin,transcription factor Snail,phosphorylated P65(p-P65),P-glycoprotein(P-gp),and programmed death-ligand 1(PD-L1)proteins in the A549 cells in various groups.Results:The Western blotting results showed that compared with M0 group,the expression levels of CD163 and Arg-1 proteins in the macrophages in M2 group were significantly increased(P<0.05),while the expression level of CD86 protein was significantly decreased(P<0.05).The immunofluorescence results showed that compared with M0 group,the expression of CD163 protein in the macrophages in M2 group was enhanced and the expression of CD86 protein was weakened.The wound healing assay results showed that at 24 and 48 h of culture,compared with A549+M0 group,the wound healing rate of the A549 cells in A549+M2 group was significantly increased(P<0.05);in the co-culture system,compared with A549+M0 group,the wound healing rate of the A549 cells in A549+M2 group was significantly increased(P<0.05);compared with A549+M2 group,the wound healing rate of the A549 cells in A549+M2+BAY11-7082 group was significantly decreased(P<0.05).The Transwell assay results showed that compared with A549+M0 group,the number of invasion A549 cells in A549+M2 group was significantly increased(P<0.05);compared with A549+M2 group,the number of invasion A549 cells in A549+M2+BAY11-7082 group was significantly decreased(P<0.05);in the co-culture system,compared with A549+M0 group,the number of invasion A549 cells in A549+M2 group was significantly increased(P<0.05).The CCK-8 assay results showed that after treated with 2.50,5.00,10.00,20.00,and 40.00 mg·L-1 DDP,compared with A549+M0 group,the inhibitory rate of proliferation of the A549 cells in A549+M2 group was significantly decreased(P<0.05 or P<0.01),and the IC50 value was significantly increased(P<0.01);in the co-culture system,compared with A549+M0 group,the inhibitory rate of proliferation of the A549 cells in A549+M2 group was significantly decreased(P<0.05 or P<0.01),and the IC50 value was significantly increased(P<0.01);compared with A549+M2 group,the inhibitory rate of proliferation of the A549 cells in A549+M2+BAY11-7082 group was significantly increased(P<0.05),and the IC50 value was significantly decreased(P<0.05).The Western blotting results showed that compared with A549+M0 group,the expression level of E-cadherin proteins in the A549 cells in A549+M2 group was significantly decreased(P<0.05),while the expression levels of N-cadherin,vimentin,and Snail proteins were significantly increased(P<0.05);in the co-culture system,compared with A549+M0 group,the expression levels of vimentin,Snail,N-cadherin,and p-P65 proteins in the A549 cells in A549+M2 group were significantly increased(P<0.05),while the expression level of E-cadherin proteins was significantly decreased(P<0.05);compared with A549+M2 group,the expression levels of vimentin,N-cadherin,and p-P65 proteins in the A549 cells in A549+M2+BAY11-7082 group were significantly decreased(P<0.05),while the expression level of E-cadherin proteins was significantly increased(P<0.05);compared with A549+M0 group,the expression levels of P-gp and PD-L1 proteins in the A549 cells in A549+M2 group were significantly increased(P<0.05);in the co-culture system,compared with A549+M0 group,the expression levels of P-gp and PD-L1 proteins in the A549 cells in A549+M2 group were significantly increased(P<0.05);compared with A549+M2 group,the expression levels of P-gp and PD-L1 proteins in the A549 cells in A549+M2+BAY11-7082 group were significantly decreased(P<0.05).Conclusion:The M2 macrophages can regulate EMT in the NSCLC cells to promote the invasion and metastasis of tumor,and modulate the expressions of P-gp and PD-L1 to enhance DDP resistance,which is associated with the NF-κB signaling pathway.

Objective:To quantitatively evaluate left ventricular myocardial work in patients with type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD) by pressure strain loop (PSL) technique.Methods:A total of 103 T2DM patients diagnosed at the Heping Hospital Affiliated to Changzhi Medical College from April 2022 to May 2023 were selected. They were divided into three groups based on their fatty liver ultrasound findings: group A (34 cases without NAFLD), group B (37 cases with mild NAFLD), and group C (32 cases with moderate to severe NAFLD). All patients underwent routine echocardiography, which measured the end diastolic left ventricular diameter (LVIDd), end systolic left ventricular diameter (LVIDs), end diastolic interventricular septum (IVSTd), left ventricular posterior wall thickness (LVPWTd), and end systolic left atrial diameter (LAD). The biplane Simpson method was used to measure left ventricular ejection fraction (LVEF), end diastolic left ventricular volume (LVEDV), and end systolic left ventricular volume (LVESV). PSL technology was applied to analyze the overall longitudinal strain (GLS), global work index (GWI), global effective work (GCW), global ineffective work (GWW), and overall work efficiency (GWE) of the left ventricular myocardium. We compared the conventional echocardiography parameters and myocardial work parameters among three groups, and analyzed the correlation between myocardial work parameters and GLS and glycosylated hemoglobin (HbA 1c). Results:The differences in body mass index (BMI), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) between group B and group A were statistically significant (all P<0.05); There was a statistically significant difference in BMI, HbA 1c, and low density lipoprotein cholesterol (LDL-C) between group C and group B (all P<0.001); The differences in BMI, HbA 1c, ALT, AST, triglyceride (TC), high density lipoprotein cholesterol (HDL-C), and LDL-C between group C and group A were statistically significant (all P<0.05). The GWI and GCW in group B were lower than those in group A (all P<0.05), while the absolute GLS, GCW, GWI, and GWE in group C were lower than those in group A, and GWW was higher than those in group A (all P<0.05); The absolute values of GLS, GCW, GWI, and GWE in group C were lower than those in group B ( P<0.05). The correlation analysis results showed that GLS and HbA 1c were negatively correlated with GCW and GWI ( r=-0.571, -0.533, -0.273, -0.223, P<0.05). Conclusions:PSL technology can quantitatively evaluate left ventricular myocardial work in patients with T2DM combined with NAFLD, and can detect early impairment of left ventricular systolic function in patients. It has clinical reference value in evaluating overall and local cardiac function.

Objective To compare the value of color Doppler echocardiography in the diagnosis of coarctation of the aorta (COA) in comparison with CT angiography (CTA). Methods Ninety-two patients with COA were retrospectively analyzed that were confirmed by surgery from January 2009 to September 2013 and diagnosed by color Doppler echocardiography and CTA. Results Among 92 COA patients conifrmed by surgery, 87 patients were diagnosed correctly by echocardiography while 91 patients were diagnosed by CTA, and the accuracy of echocardiography and CTA were 95%(87/92) and 99%(91/92), respectively without significant difference (χ2=1.550, P=0.213). According to surgical findings, there were 222 anatomic abnormalities including 110 intra-cardiac, 13 heart-vessel connection and 99 vessel abnormalities. For 110 intra-cardiac abnormalities, echocardiography detected all abnormalities while CTA found 92 with 18 missed including 11 valve or valve ring lesions, 3 patent foramen ovale, 2 atrial septal defect, 1 ventricular septal defect and 1 endocardial cushion defect. The accuracy of echocardiography for intra-cardiac abnormalities was signiifcantly higher than that of CTA (100%, 110/110 vs 84%, 92/110,χ2=18.801, P=0.000). CTA and echocardiography detected all abnormalities of heart-vessel connection. For 99 vessel abnormalities, CTA showed an accuracy of 98%(97/99) with 2 missed lesions including 1 collateral circulation between the aorta and the pulmonary artery and 1 membranous narrow of the aorta, while echocardiography misdiagnosed 1 COA as interruption of aortic arch and missed 15 lesions, including 4 COA, 3 collateral arteries, 2 patent ductus arteriosus, 2 aberrant right subclavian artery, 2 brachiocephalic vein abnormality, 1 partial anomalous pulmonary venous drainage and 1 persistent left superior vena cava. The accuracy of CTA for detection of vessel abnormalities was signiifcantly higher than that of echocardiography (98%, 97/99 vs 84%, 83/99,χ2=11.350, P=0.000). Conclusions Both CTA and echocardiography are effective diagnostic modalities for COA and CTA is superior to echocardiography for detecting vessel anomalies whereas echocardiography is superior to CTA for detecting intra-cardiac abnormalities. The combination of both modalities will provide more accurate information for clinical diagnosis.

Objective To establish an isolation method for solid GTC in peripheral blood using EpCAM antibody-linked nanobeads and evaluate the sensitivity of the method and its application significance. Methods Five, ten, twenty, fifty and one hundred MCF7 (breast cancer), KYSE70 (esophageal cancer), BxPC-3 (pancreatic cancer) and 9811P (stomach cancer) cells were added into 7. 5 ml erythrocyte lysed peripheral blood obtained from healthy volunteers respectively. EpCAM antibodylinked nanobeads were used to enrich cancer cells. The recovery rates of the in vitro added cancer cells were evaluated by fluorescence microscopy. Then, the untreated thirty cases of esophageal cancer (six cases at stage Ⅰ and Ⅱ, twenty-four cases at stage Ⅲ and Ⅳ), thirty-five cases of breast cancer (fifteen cases at stage Ⅰ and Ⅱ , twenty cases at stage Ⅲ and Ⅳ), thirty cases of pancreatic cancer (five cases at stage Ⅰ and Ⅱ , twenty-five cases at stage Ⅲ and Ⅳ), thirty-three gastric cancer (thirteen cases for stage Ⅰ and Ⅱ ,twenty cases at stage Ⅲ and Ⅳ) were enrolled to enrich the peripheral blood CTC. Thirty healthy volunteers and thirty gastritis patients served as two groups of control. Meanwhile the enriched CTC was identified by IF and HE staining. FISH was used to analyze the copy number of chromosome 8 and chromosome 20 in two hundred esophageal cancer, breast cancer, pancreatic caner and gastric cancer CTC. Results After DAPI staining and mixing with 7.5 ml peripheral blood from healthy donors, the average cell recovery rates of KYSE70, MCF7, BxPC-3 and 9811P cells evaluated under fluorescence microscope were 87%, 87%, 86% and 88% (within group), and the recovery rates of 5 gradient dilution levels were 88%, 85%, 87%, 88% and 87% (intergroup). With a high sensitivity, this method was able to isolate one cancer cell in 107 white blood cells of peripheral blood. The positive rates of more than 2 CTC in the peripheral blood detected by this method were 50% (15/30) of esophageal cancer, 63% (22/35) of breast cancer, 70% (21/30) of pancreatic cancer and 61% (20/33) gastric cancer patients respectively,but no CTC was detected in the peripheral blood of healthy volunteers and gastritis patients (P = 0. 000).The aneusomy of chromosome 8 and chromosome 20 were found in 80% esophageal cancer, 75% breast cancer, 65% pancreatic cancer and 59% gastric cancer. Conclusions The CTC isolation technique with EpCAM antibody-linked nanobeads is sensitive and accurate. The aneusomy of chromosome 8 and 20 is frequent in CTC from esophageal cancer, breast cancer, pancreatic cancer and gastric cancer.

Anopheles minimus was infected with Plasmodium yoelii yoelii under adequate condition. The malaria parasite could develop normally and produce mature oocysts containing sporozoites. Large numbers of free active sporozoites were found in the haemocele but most of them failed to penetrate into the mosquito's salivary glands. Thus the sporogonic cycle of the parasite could not be completed.But in one mosquito, a few sporozoites were found in its salivary glands. It seems that there is an individual difference in the susceptibility of the mosquitoes and this throws some light on the possibility of selecting a susceptible strain of A. minimus to establish the A. minimus-P. yoelii system for malaria research.