Objective To investigate the correlation between the clinical pathological factors and the expression of EGFR, HER2 and HER3 in non-small cell lung cancer tissues. Methods The expression of EGFR, HER2 and HER3 in non-small cell lung cancer tissues was detected by MaxVision immunohistochemical. The relationship between the protein expression and the clinical pathologic characteristics was studied. The correlation between the protein expression and the survival was also retrospectively analyzed. Results Increased expression of EGFR, HER2 and HER3 was detected in the NSCLC tissues and their rates were 45.9 % (73/156), 30.8 % (49/156) and 21.4 % (37/156), respectively. There were significant associations between the expression of these proteins and lymph node metastasis, tumor differentiation degree and TNM staging. Conclusions Overexpression of EGFR, HER2, HER3 or all three proteins is a predictor of poor prognosis. Moreover, combined detection of all 3 markers may work better than the individual detection in the prediction of the prognosis to guide the clinical treatment.

BACKGROUND:The repair method of proximal humeral comminuted fractures in the elderly remains controversial. OBJECTIVE:To compare the recovery of shoulder joint function using the hemiarthroplasty of shoulder and locking proximal humeral plate fixation for complex proximal humeral fracture in the elderly. METHODS:A total of 22 elderly patients with complex proximal humeral fracture, who were treated from October 2012 to October 2014, were retrospectively analyzed. 12 cases underwent hemiarthroplasty, and 10 cases received locking proximal humeral plate fixation. Al patients were folowed up after treatment. X-ray films were rechecked during the folow-up. The recovery of shoulder function was evaluated with Neer score. RESULTS AND CONCLUSION:During final folow-up, no prosthetic loosening or avascular necrosis of humeral head was found in the shoulder hemiarthroplasty group, and the Neer score was (81±5) points. In the locking proximal humeral plate fixation group, there was fixator loosening in four cases and avascular necrosis of humeral head in three cases, and the Neer score was (69±5) points (P < 0.05). During repair, mean operation time was shorter in the shoulder hemiarthroplasty group than in the locking proximal humeral plate fixation group. The amount of bleeding was higher in the shoulder hemiarthroplasty group than in the locking proximal humeral plate fixation group (P > 0.05). These data suggest that locking proximal humeral plate fixation and shoulder hemiarthroplasty are effective repair methods for complex proximal humerus fractures, but the hemiarthroplasty had the advantage of early functional exercise and good recovery of shoulder joint.

Present study was focused on the chemical constituents of the stems and leaves of Salvia yunnanensis C . H. Wright and their anti-angiogeneic activities. The compounds were isolated by column chromatography over silica gel and Sephadex LH-20, and other isolation techniques. Their structures were elucidated on the basis of spectral analysis and chemical evidences. Their anti-angiogeneic activities were evaluated by the chicken chorioallantoic membrane (CAM) neovascularisation model. Seven compounds were separated and identified as ( + ) -spathulenol( 1), 5,7,4'-trihydroxyflavanone(2) , beta-amyrin(3), 3 beta-hydroxy-12-ursene(4), 2alpha,3 beta-dihydroxyursa-12-en-28-oic acid(5), ursolic acid (6) and 3-oxo-12-ursen-28-oic acid (7). Compounds 1, 2, 5 and 6 were obtained from this plant for the first time. Compounds 5 (an oleanane compound) and 6 (an ursane compound) could inhibit angiogenesis significantly in a dose-dependent manner.
Angiogenesis Inhibitors ; chemistry ; pharmacology ; Animals ; Chorioallantoic Membrane ; drug effects ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Salvia ; chemistry

Objective:To investigate the correlation of CD8 positive tumor-infiltrating lymphocytes (CD8 + TIL) density and programmed-death receptor ligand 1 (PD-L1) expression in rectal cancer with clinicopathological characteristics and prognosis of patients after neoadjuvant chemoradiotherapy. Methods:The clinicopathological data of 166 patients with locally advanced rectal cancer (LARC) who received neoadjuvant therapy before surgery in the Beijing Chao-Yang Hospital, Capital Medical University from January 2015 to December 2018 were retrospectively analyzed. CD8 + TIL density and PD-L1 expression were detected by using immunohistochemistry. The correlation of CD8 + TIL density and PD-L1 expression with clinicopathological characteristics of patients after neoadjuvant chemoradiotherapy was analyzed. Kaplan-Meier method was used to analyze the disease-free survival (DFS) and Cox regression risk model was used to make univariate and multivariate analysis of the influencing factors for DFS. Results:Among 166 LARC patients, 81 cases (48.8%) had high density of CD8 + TIL, 85 cases (51.2%) had low density of CD8 + TIL; 63 cases (38.0%) had PD-L1 expression, and 103 cases (62.0%) had non-expression of CD8 + TIL. The expression rate of PD-L1 in CD8 + TIL high density group was higher than that in CD8 + TIL low density group [50.6% (41/81) vs. 25.9%(22/85), χ2 = 10.78, P < 0.001]. According to the density of CD8 + TIL and PD-L1 expression, immunophenotype was divided among 4 groups; the 3-year DFS rate of the CD8 + TIL high density /PD-L1 expression group was 87.1%, which was higher than that of the other groups (CD8 + TIL low density /PD-L1 expression group was 72.8%, CD8 + TIL high density /PD-L1 non-expression group was 67.0%, CD8 + TIL low density /PD-L1 non-expression group was 64.3%), and the difference was statistically significant ( P < 0.05). Univariate analysis showed that tumor differentiation degree, TNM stage, CD8 + TIL density, PD-L1 expression and CD8 + TIL density /PD-L1 expression were correlated with the DFS of patients (all P < 0.05). Multivariate analysis results showed that CD8 + TIL high density /PD-L1 expression was an independent protective factor for DFS ( HR = 0.049, 95% CI 0.005-0.497, P = 0.011), while TNM stage 3 was an independent risk factor for DFS ( HR = 2.752,95% CI 1.300-5.825, P = 0.008). Conclusions:In LARC after neoadjuvant therapy, CD8 + TIL density is positively correlated with the expression of PD-L1, and the high density of CD8 + TIL/PD-L1 expression is an independent influencing factor for good prognosis, suggesting that these patients may benefit from the immunotherapy.

Objective:To explore the effect of peripheral blood T lymphocyte subsets on locally advanced rectal cancer after neoadjuvant chemoradiotherapy.Methods:108 patients were included at the Department of General Surgery of Beijing Chaoyang Hospital from Jun 2016 to Jun 2017. Peripheral blood was collected within one week before neoadjuvant therapy and one week before rectal surgery. Flow cytometry was applied to detect the CD3 + 、CD4 + 、CD8 + 、CD45RA + 、CD45RO + expression. Receiver operating characteristic (ROC) curve analysis was performed to determine the best cut-off value of the ratio of lymphocytes. A logistic regression model was obtained in multivariate analysis. Results:The values of CD3 + , CD4 + and CD8 + T lymphocytes in peripheral blood of the patients decreased compared with that before neoadjuvant treatment (all P<0.05). There was no significant decrease in the proportion of CD4 + , CD8 + , CD45RA + T and CD45RO + lymphocytes in patients′ peripheral blood (all P>0.05). The CD45RO in peripheral blood decreases during neoadjuvant therapy for locally advanced rectal cancer, and it is associated with better tumor regression( P<0.05). The best cut-off value for the ratio changes of CD45RO was 1.07. The ratio changes of CD45RO were the only significant factor for tumor regression in multivariate analysis ( P=0.005, OR=26.867, 95% CI: 1.530-471.635). Conclusion:The percentage of peripheral blood CD45RO may predict the sensitivity of neoadjuvant therapy in rectal cancer patients.

Objective To investigate the correlation between serum miR-320b and carotid atherosclerosis in patients with acute ischemic stroke.Methods From January 2017 to December 2017,patients with acute ischemic stroke visited the Department of Neurology,the Affiliated Hospital of Yangzhou University were enrolled.According to the findings of carotid artery ultrasonography,they were divided into plaque group and plaque-free group.The baseline clinical data such as demographic data,vascular risk factors,and blood biochemical indicators were collected.Reverse transcription quantitative polymerase chain reaction was used to detect the expression level of serum miR-320b.Multivariatelogistic regression analysis was used to determine the independent risk factors for carotid atherosclerosis.Results A total of 135 patients with acute ischemic stroke were enrolled in this study,including 58 females and 77 males,aged 58.4 ± 10.6 years.There were 85 patients in the plaque group and 50 in the plaque-free group.The total cholesterol (t =5.523,P =0.023) and low-density lipoprotein cholesterol (t =4.415,P =0.044) in the plaque group were significantly higher than those in the plaque-free group,while high-density lipoprotein cholesterol (t =5.849,P=0.017) and serum miR-320b (t =4.331,P=0.039) were significantly lower than those in the plaque-free group.Multivariate logistic regression analysis showed that referring to the highest quartile group,the low serum miR-320b level might be an independent risk factor for carotid atherosclerosis (the first quartile group:odds ratio 2.701,95％ confidence interval 1.154-6.321,P =0.022;the second quartile group:odds ratio 2.521,95％ confidence interval 1.249-5.091,P =0.010;and the third quartile group:odds ratio 1.849,95％ confidence interval 1.041-3.283,P=0.036).Conclusion The low serum miR-320b level might be an independent risk factor for carotid atherosclerosis in patients with acute ischemic stroke.

Objective:To compare the short-term efficacy and perioperative safety of neoadjuvant chemoradiotherapy (nCRT) with total neoadjuvant treatment (TNT) in patients with locally advanced rectal cancer (LARC).Methods:A retrospective cohort analysis was carried out. Inclusion criteria: (1) rectal adenocarcinoma confirmed by pathology with a distance from tumor inferior border to anal verge within 12 cm; (2) clinical stage cT3-4N0 or cT1-4N1-2 diagnosed by magnetic resonance imaging (MRI) or endorectal ultrasonography; (3) a single rectal tumor confirmed by colonoscopy; (4) patients suitable for chemoradiotherapy; (5) no previous history of other tumors. Exclusion criteria: (1)patients with previous rectal cancer surgery and local recurrence; (2) those who did not complete nCRT course; (3) those with distant metastases; (4) those with defective clinicopathological data. According to the above criteria, a total of 134 LARC patients at the Department of General Surgery of Beijing Chaoyang Hospital from January 2016 to January 2019 were enrolled, including 82 males and 52 females, with a male-female ratio of 1.58∶1.00 and mean age of (59.6±11.2) (26-81) years. Based on neoadjuvant regimen, patients were divided into nCRT group ( n=55) and TNT group ( n=79). There were no statistically significant differences in baseline data, such as age, sex, distance from tumor to anal verge, Eastern Cooperative Oncology Group (ECOG) performance status and clinical TNM stage, between the two groups (all P>0.05). All the patients received pelvic intensity-modulated radiotherapy (IMRT) with a total dose of 50.4 Gy in 28 fractions. Patients in nCRT group received oral capecitabine chemotherapy during radiotherapy and underwent surgery 6-8 weeks after chemoradiation. Patients in TNT group received one cycle of induction CapeOX (oxaliplatin and capecitabine) and concurrent chemoradiotherapy, then underwent a radical surgery two weeks after completion of consolidation chemotherapy. The efficacy of neoadjuvant therapy, adverse events of chemoradiotherapy and perioperative safety were compared between the two groups. Results:Patients of two groups completed the course of neoadjuvant therapy. There were no statistically significant differences between nCRT group and TNT group in the incidence of adverse events in neutropenia [7.3% (4/55) vs. 10.1% (8/79)], anemia [3.6% (2/55) vs. 3.8% (3/79)], thrombocytopenia [5.5% (3/55) vs. 7.6% (6/79)], gastrointestinal dysfunction [3.6% (2/55) vs. 6.3% (5/79)] and radiation enteritis [9.1% (5/55) vs. 8.9% (7/79)] (all P>0.05). One hundred and thirty patients completed TME surgery, including 54 patients in nCRT group and 76 patients in the TNT group. Compared with the nCRT group, the proportion of abdominoperineal resection (APR) was higher in the TNT group [31.6% (25/76) vs. 13.0% (7/54), χ 2=9.382, P=0.009]. No statistically significant differences in morbidity of postoperative complication, operation time, intraoperative blood loss and postoperative hospital stay between the two groups were found (all P>0.05). The distal and circumferential margins were negative in all the patients. Seventeen patients in the TNT group 22.4% (17/76) got pathologic complete response (pCR), which was significantly higher than 7.4% (4/54) in nCRT group (χ 2=5.217, P=0.022). There were no statistically significant differences in ypTNM classification, perineural invasion and venous invasion between the two groups (all P>0.05). Conclusion:The pCR of TNT is higher than that of nCRT without increasing the incidence of toxicity and complications of radiotherapy and chemotherapy for patients with locally advanced rectal cancer.

Objective:To compare the short-term efficacy and perioperative safety of neoadjuvant chemoradiotherapy (nCRT) with total neoadjuvant treatment (TNT) in patients with locally advanced rectal cancer (LARC).Methods:A retrospective cohort analysis was carried out. Inclusion criteria: (1) rectal adenocarcinoma confirmed by pathology with a distance from tumor inferior border to anal verge within 12 cm; (2) clinical stage cT3-4N0 or cT1-4N1-2 diagnosed by magnetic resonance imaging (MRI) or endorectal ultrasonography; (3) a single rectal tumor confirmed by colonoscopy; (4) patients suitable for chemoradiotherapy; (5) no previous history of other tumors. Exclusion criteria: (1)patients with previous rectal cancer surgery and local recurrence; (2) those who did not complete nCRT course; (3) those with distant metastases; (4) those with defective clinicopathological data. According to the above criteria, a total of 134 LARC patients at the Department of General Surgery of Beijing Chaoyang Hospital from January 2016 to January 2019 were enrolled, including 82 males and 52 females, with a male-female ratio of 1.58∶1.00 and mean age of (59.6±11.2) (26-81) years. Based on neoadjuvant regimen, patients were divided into nCRT group ( n=55) and TNT group ( n=79). There were no statistically significant differences in baseline data, such as age, sex, distance from tumor to anal verge, Eastern Cooperative Oncology Group (ECOG) performance status and clinical TNM stage, between the two groups (all P>0.05). All the patients received pelvic intensity-modulated radiotherapy (IMRT) with a total dose of 50.4 Gy in 28 fractions. Patients in nCRT group received oral capecitabine chemotherapy during radiotherapy and underwent surgery 6-8 weeks after chemoradiation. Patients in TNT group received one cycle of induction CapeOX (oxaliplatin and capecitabine) and concurrent chemoradiotherapy, then underwent a radical surgery two weeks after completion of consolidation chemotherapy. The efficacy of neoadjuvant therapy, adverse events of chemoradiotherapy and perioperative safety were compared between the two groups. Results:Patients of two groups completed the course of neoadjuvant therapy. There were no statistically significant differences between nCRT group and TNT group in the incidence of adverse events in neutropenia [7.3% (4/55) vs. 10.1% (8/79)], anemia [3.6% (2/55) vs. 3.8% (3/79)], thrombocytopenia [5.5% (3/55) vs. 7.6% (6/79)], gastrointestinal dysfunction [3.6% (2/55) vs. 6.3% (5/79)] and radiation enteritis [9.1% (5/55) vs. 8.9% (7/79)] (all P>0.05). One hundred and thirty patients completed TME surgery, including 54 patients in nCRT group and 76 patients in the TNT group. Compared with the nCRT group, the proportion of abdominoperineal resection (APR) was higher in the TNT group [31.6% (25/76) vs. 13.0% (7/54), χ 2=9.382, P=0.009]. No statistically significant differences in morbidity of postoperative complication, operation time, intraoperative blood loss and postoperative hospital stay between the two groups were found (all P>0.05). The distal and circumferential margins were negative in all the patients. Seventeen patients in the TNT group 22.4% (17/76) got pathologic complete response (pCR), which was significantly higher than 7.4% (4/54) in nCRT group (χ 2=5.217, P=0.022). There were no statistically significant differences in ypTNM classification, perineural invasion and venous invasion between the two groups (all P>0.05). Conclusion:The pCR of TNT is higher than that of nCRT without increasing the incidence of toxicity and complications of radiotherapy and chemotherapy for patients with locally advanced rectal cancer.

To establish a rabbit model of proprotein convertase subtilisin/kexin type9 () point mutation with CRISPR/Cas9 gene editing technique. According to the PubMed gene protein data, the PCSK9 protein functional regions of human and rabbit were analyzed by Blast. The 386S (Ser) amino acid functional region of human gene was homologous to the 485S of rabbit gene. Three small guide RNAs and one single-stranded donor oligonucleotide were designed according to the 485S base substitution position and sequence analysis of rabbit gene. The synthetic small guide RNAs, Cas9 mRNA and single-stranded donor oligonucleotide were co-injected into the cytoplasm of rabbit fertilized eggs and the embryos were transferred into the pregnant rabbits. PCR, TA cloning and off-target analysis were performed on the F0 rabbits to identify whether the PCSK9 mutation was successful. Fifteen F0 rabbits were obtained. The sequencing results showed that one of them was PCSK9 point mutation homozygote and two of them were PCSK9 point mutation heterozygotes, and the mutation could be stably inherited. The rabbit model of PCSK9 point mutation was successfully constructed by CRISPR/Cas9 technique, which provides an animal model for exploring the molecular mechanism of impaired PCSK9 function and developing reliable and effective diagnosis and treatment measures.
Animals ; CRISPR-Cas Systems/genetics* ; Mutation ; Point Mutation ; Proprotein Convertase 9/metabolism* ; Rabbits

Objective:To investigate the effect and mechanism of the gastric cancer cells-derived exosome miR-382-5p in-duced by Helicobacter pylori(H.pylori)on the autophagy and polarization of macrophages,providing new clues for further elucidating the carcinogenic mechanism of H.pylori.Methods:Ultracentrifugation and exosome extraction kit were used to extract the exosomes re-leased by the H.pylori stimulated group and the blank control group AGS cells cells,then transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA)and Western blot were employed to identify exosomes.qRT-PCR was used to detect the expres-sion of miR-382-5p in H.pylori induced AGS-derived exosomes.miR-382-5p mimic was transfected into THP-1 macrophages,then the expressions of autophagy markers(LC3Ⅱ,p62,and Beclin-1)were evaluated by Western blot,the number of autophagosomes was detected by immunofluorescence.The expression levels of PTEN protein,downstream proteins PI3K,AKT,mTOR and its phosphory-lated proteins p-PI3K,p-AKT,p-mTOR were detected by Western blot.Flow cytometry was used to detect the expression levels of macrophage phenotypic molecules CD206 and HLA-DR.ELISA was used to detect the secretion of cytokines TNF-α,IL-6,IL-10 and Arginase1 in macrophage supernatants.Results:The extracted exosomes were consistent with exosome morphology and highly ex-pressed the surface marker proteins CD9,CD63 and TSG101.Compared with the blank control group,the expression level of exosom-al miR-382-5p in H.pylori-infected group was significantly increased.miR-382-5p mimic transfection resulted in decreased expression of LC3 Ⅱ and Beclin-1 in macrophages,increased expression of P62 and decreased number of autophagosomes.Moreover,the protein expression level of PTEN was significantly decreased in the miR-382-5p mimic transfection group,while the expression levels of p-PI3K,p-AKT and p-mTOR were significantly increased.miR-382-5p mimic transfection also resulted in increased expression of mac-rophage M2 type marker protein CD206 and decreased expression of M1 type marker protein HLA-DR,as well as increased expres-sions of IL-10 and Arginine1,whereas decreased expression of IL-6 and TNF-α.Pretreatment with the pathway inhibitor BEZ235 par-tially reverses the effects of miR-382-5p on macrophage autophagy and polarization.Conclusion:H.pylori-induced gastric cancer cells-derived exosomal miR-382-5p suppresses macrophage autophagy and induces M2 polarization through down-regulation of PTEN ex-pression and activation of the PI3K/AKT/mTOR signaling pathway.