AIM:To synthesize a safe , efficient and targeted nanoparticulate carrier for siRNA delivery to pan-creatic cancer cells .METHODS: Iron oxide nanocrystal with carboxylic acid group-polyethyleneimine ( IONP-PEI ) was synthesized and investigated as a nonviral carrier of siRNA to the pancreatic cells .The size, surface and charge using zeta potential were characterized .The perfect charge ratio between amino groups of IONP-PEI and phosphate groups of siRNA ( N/P) was determined by the transfection efficiency detection , gel retardation assay and MTS assay .An antibody-directed nonviral vector , scFvCD44v6-IONP-PEI nanoparticle attaching to the cancer-associated CD44v6 single-chain variable frag-ment, was constructed as a cancer-targeting nanocarrier for siRNA delivery .Prussian blue staining and immunofluorescent staining were performed to detect the distribution of scFv CD44v6-IONP-PEI/siRNA complexes in the cells .The transfection efficiency , fluorescence intensity and the expression of KRAS at mRNA and protein levels in the cells transfected by IONP -PEI/siRNA and scFv CD44v6-IONP-PEI/siRNA were detected by flow cytometry , fluorescence microscopy , real-time PCR and Western blotting, respectively.RESULTS:The mass ratio of IONP to PEI was 0.75.The suitable ratio of N/P was 20. The averaged size and surface zeta potential of IONP-PEI/siRNA in deionized water were (51.3 ±2.2)nm (diameter) and (21.73 ±8.07)mV, respectively.Red fluorescence was seen in both targeting and nontargeting groups , which clearly re-vealed the intracellular distribution of siRNA and delivery agents .Transfection efficiencies in targeting and nontargeting groups were (89.75 ±1.81)%and (59.87 ±4.52)%, respectively.Down-regulation of the KRAS mRNA in Panc-1 cells transfected with siKRAS by scFvCD44v6-IONP-PEI and IONP-PEI was up to (34.02 ±6.15)%and (51.09 ±6.70)%, re-spectively .The protein level of KRAS was lower in targeting group than that in nontargeting group .CONCLUSION:scFvCD44v6-IONP-PEI is a safe and efficient nanoparticulate carrier for gene delivery .It is more effective to transfer siRNA into the cells and mediate gene silencing effect in vitro than the nontargeting group .

Objective To compare the clinical application of Da Vinci robot surgical system (RSS) with traditional open surgery (TOS).Methods From Feb 2015 to Jul 2016,48 cases of upper abdominal surgical disease patients were divided into RSS group (23 cases) and TOS group (25 patients) randomly.Results The anesthesia time [(194 ±16)min vs.(181 ±11)min,t=3.262,P=0.002] and operation time [(167 ± 14) min vs.(158 ± 14) min,t =2.292,P =0.027] were much longer in the RSS,while the blood loss during operation significantly less than the TOS [(128 ± 62) ml vs.(190 ± 86) ml,t =-2.886,P =0.006].The RSS has obvious advantages in 24 h-drainage [(69 ± 27) ml vs.(114 ± 54) ml.t =-3.680,P =0.001],time to out-of-bed activity [(27.7 ± 8.0) h vs.(35.7 ± 9.9) h,t =-3.067,P =0.004],BPS,postoperative exhausting time[(27.2 ±5.9)h vs.(32.8 ±8.3)h,t =-2.690,P =0.01] and length of hospital stay [(10.4 ± 1.8) d vs.(11.8 ± 1.9) d,t =-2.600,P =0.013].But the total hospital cost was higher in RSS [(117 000 ± 10 000) yuan vs.(77 000 ± 8 000) yuan,t =15.087,P =0.000)].Conclusions The RSS is a much minimally invasive surgery,reducing blood loss and postoperative pain,promoting rapid recovery,shortening hospital stay.

Objective To investigate the expression of PD-1 on CD4 +and CD8 + T cells in patients with HBeAg-positive chronic hepatitis B(CHB)treated with telbivudine.Methods Fifty-six HBeAg-positive CHB patients admitted in the First Affiliated Hospital of Zhengzhou University during January 201 3 and June 201 4 were enrolled in this study.The expression of PD-1 on CD4 +and CD8 + T cells was detected with flow cytometry at baseline,24,48 and 72 wks after telbivudine treatment.The relationship of PD-1 expression with alanine aminotransferase (ALT)level,HBeAg seroconversion and HBV DNA loads was analyzed.t test and completely random variance analysis were used to analyze the data.Results The PD-1 expression on CD4 + and CD8 + T cells at baseline was higher in patients with low ALT levels compared to those with high ALT levels(t =1 2.20 and 9.69,both P <0.01 ),while higher levels of PD-1 expression was also observed in patients with high HBV DNA load (≥5 lgIU /mL)compared to those with low HBV DNA load (t =4.39 and 4.85,both P <0.01 ).PD-1 levels on CD4 + and CD8 + T cells presented a declining trend after telbivudine treatment(F =6.98 and 8.97,both P <0.01 ),PD-1 expression in patients with HBeAg seroconversion showed lower levels compared with baseline values (t =1 8.45 and 1 8.01 ,both P <0.01 ). Conclusion In HBeAg-positive CHB patients,the expression of PD-1 on CD4 + and CD8 + T lymphocytes shows a decreasing trend during the treatment with telbivudine,indicating that antiviral therapy may alleviate the immunosuppression in these patients.

Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.

Objective To evaluate the effect of methylene blue (MB) on hydrogen peroxide (H2O2)-induced apoptosis in macrophages through mitochondria-dependent pathway in mice.Methods Mouse peritoneal macrophage line RAW264.7 cells were cultured in DMEM culture medium containing 10％ fetal bovine serum.Cells were divided into 6 groups (n =24 each) using a random number table method:control group (group C),H2O2 group,prophylactic different concentrations of MB groups (MB1,2 groups) and therapeutic different concentrations of MB groups (MB3.4 groups).H2O2 50 μmol/L was added to the culture medium in group H2O2.MB was added to the culture medium with the final concentrations of 0.1 μmol/L (in MB1 and MB3 groups) and 1.0 μmol/L (in MB2 and MB4 groups) at 30 min before adding H2O2 in MB1.2 groups and 30 min after adding H2O2 in MB3.4 groups.At 24 h of culture or incubation in each group,the cell survival rate was measured by methyl thiazolyl tetrazolium assay,the activity of reactive oxygen species (ROS) in cells was determined with the fluorescent probe,the lactate dehydrogenase (LDH) activity in supernatant was detected by spectrophotometry,the activity of superoxide dismutase (SOD) in cells was detected by colorimetric method,mitochondrial membrane potential (MMP) was measured using rhodamine 123 staining,the content of ATP was determined by an ATP bioluminescent method,the expression of pro-caspase-3 and spliceosomes P20 protein and P 18 protein was detected by Western blot,and cell apoptosis was detected using flow cytometry.Results Compared with group C,the cell survival rate,SOD activity and contents of MMP and ATP were significantly decreased,the ROS activity and activity of LDH in supernatant were increased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was up-regulated,and early and late apoptosis rates were increased in the other five groups (P＜0.05).Compared with group H2O2,the cell survival rate,SOD activity and contents of MMP and ATP were significantly increased,the ROS activity and activity of LDH in supernatant were decreased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was down-regulated,and early and late apoptosis rates were decreased in MB1-4 groups (P＜0.05).Compared with group MB1,the cell survival rate was significantly decreased,and the expression of caspase-3 spliceosome P 18 was down-regulated in group MB2,and the cell survival rate and SOD activity were significantly decreased,and the activity of ROS was increased in group MB3 (P＜0.05).Compared with group MB4,the expression of caspase-3 spliceosome P 18 was significantly down-regulated,early and late apoptosis rates were decreased,and the activity of ROS was increased in group MB2,and the activity of ROS was significantly increased in group MB3 (P＜0.05).Conclusion The mechanism by which MB attenuates H2O2-induced oxidative damage to macrophages is related to inhibiting cell apoptosis in macrophages through mitochondria-dependent pathway in mice.

Objective:To discuss the principle and effect of augmentation rhinoplasty with auricular cartilage and expanded polytetrafluoroethylene.Methods:From January 2018 to January 2020, 161 patients (10 males and 151 females; aged from 19 to 48 years, with an average of 26 years) underwent " auricular cartilage plus expanded polytetrafluoroethylene" augmentation rhinoplasty in Nanfang Hospital of Southern Medical University. The expended polytetrafluoroethylene was carved into a willow leaf shape (I Shape) to fill the nasal dorsum, and the cartilage taking from cymba concha was constructed into an arched bridge shape for the nasal tip shaping. Pre-operative and 1-year post-operative measurements nasal length, nasal height, nasal depth, nasal columella height, nasal tip width, nasofrontal angle, nasolabial angle, survey of satisfaction and complication rate 1-2 years after operation were taken. The statistical analysis of nasal morphological indicators and nasal aesthetic indicators were employed.Results:The nose shape of 161 patients was improved to varying degree. All morphological indicators were improved, and difference was statistically significant ( P<0.05). The nasofrontal angle reached the standard in 90 cases, accounting for 55.9%; The nasolabial angle reached the standard in 143 cases, accounting for 88.8%. 2 cases had prosthesis (ePTFE) deviation and were corrected by surgical repair; 1 case had prosthesis (ePTFE) rejection and was corrected by prosthesis (ePTFE) removal surgery. Conclusions:Corresponding to the anatomical characteristics of the external nose, the prosthesis material is designed and made to correspond to the dorsum shape of the nasal stent. The shape of the alar cartilage, the prefabricated arched bridge shape of the cymba concha cartilage are used to reconstruct the nasal tip, which can effectively elevate the nasal dorsum, improve the protruding degree and rotation degree of the nasal tip, and have good long-term support. The flexibility and activity of the nasal tip are similar to the biological nose.

Objective: To explore the association between urinary metals and lung function among college students, and to provide a theoretical basis for related research on metal exposure and lung function injury. Methods: A total of 45 healthy college students were recruited from North China University of Science and Technology in Caofeidian between 2017-2018. During the four seasons, information was obtained from questionnaires and physical examinations, lung function parameters were assessed, including FVC, FEV1, PEF, FEV1/FVC and FEF 25-75 , and morning urine samples were collected simultaneously. The urinary levels of 15 metals were measured by inductively coupled plasma mass spectrometry (ICP/MS); a Kruskal Wallis H test was used to compare differences in urinary metals during the four seasons; and a mixed effect model was used to assess correlations between urinary metals and lung function. Results: There were significant differences in the levels of urinary chromium, iron, nickel, copper, zinc, arsenic, selenium, selenium, molybdenum, cadmium, antimony and lead from 15 metals over the four seasons ( H =9.79- 20.61 , P <0.05). The differences observed in five lung function parameters over the four seasons were statistically significant ( F =61.72, 45.30, 47.61, 25.47, 35.13, P <0.05). The linear mixed effect model analysis showed that urinary concentrations of vanadium, manganese, iron, cobalt, nickel and antimony were negatively correlated with FEV1( B =0.202, 0.192, 0.181, 0.154, 0.131 , 0.283); urinary concentrations of aluminum, vanadium, manganese, iron, cobalt, nickel, zinc, cadmium, and antimony were negatively correlated with FVC ( B =0.252, 0.290, 0.292, 0.271, 0.201, 0.180, 0.171, 0.163, 0.381); urinary concentrations of manganese and antimony were negatively correlated with PEF ( B =0.291, 0.354)( P <0.05). Conclusion The increase of multiple metal concentrations among college students was related to lung function decline, the long term metal exposure might lead to lung function damage. So environmental metal pollution should be controlled.

NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome mediates inflammation, induces pyroptosis, and regulates periodontal tissue remodeling through the maturation and secretion of its downstream cysteine protease 1 (Caspase-1)-dependent pro-inflammatory cytokines, interleukin (IL)-1β and IL-18. Orthodontic force mediates the aseptic inflammation of periodontal tissues and triggers adaptive alteration of periodontal tissues, thereby promoting the movement and stability of orthodontic teeth. NLRP3 inflammasome plays an important role in orthodontic tooth movement and causes periodontal tissue inflammation and orthodontic inflammatory root resorption in orthodontic patients. Literature review suggests that NLRP3 inflammasome is involved in the activation and differentiation of periodontal ligament fibroblasts, periodontal ligament stem cells, macrophages, osteoblasts, and osteoclasts in orthodontic tooth mobile tissue remodeling. Additionally, it targets the upstream nuclear factor kappa-B signaling pathway; downstream effectors, such as Caspase-1, IL-1β, and IL-18; and the NLRP3 inflammasome components for regulating tooth movement as well as treating and preventing orthodontics-associated periodontitis and orthodontic-induced inflammatory root resorption. Future studies can be focused on the specific mechanism of NLRP3 inflammasome tissue modification during orthodontic tooth movement. This article reviews the effects and regulatory mechanisms of the NLRP3 inflammasome signaling pathway on the corresponding tissue remodeling during orthodontic tooth movement.

Objective:To systematically retrieve the assessment tools for patient care complexity at home and abroad, and compare their release institutions, formation process, applicable population, assessment methods and main contents.Methods:Articles on patient care complexity assessment tools was retrieved through computers in PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure, WanFang Data, China Biomedical Database, and VIP Database. The retrieval time limit was from the establishment of the database to July 2, 2022. Excel 2019 was used to extract the name, release institution, evaluation method, main content and other information of the assessment tools and compare the content characteristics of the assessment tools.Results:A total of 26 assessment tools for patient care complexity were included in this study, involving 37 articles, mainly observational studies. The 26 tools were mainly issued by institutions of higher learning, and most of them were applicable to ordinary inpatients. The assessment contents included the general situation of the patient, severity of the disease, treatment items, mental cognitive function, mental health, socio-economic support and compliance.Conclusions:At home and abroad, the patient care complexity evaluation tools are mainly used to evaluate ordinary adult inpatients. There are few tools to evaluate children, the elderly, Intensive Care Unit (ICU) and other special types of patients, and there is a lack of relevant intervention research. In China, there are few tools for evaluating patient care complexity involving indicators related to mental cognitive function, which suggests that nurses can consider the mental cognitive function of patients when building relevant assessment tools in the future. In addition, the application effect of such tools in patient risk identification and safety management, nursing human resource allocation and other fields should be verified.

Objective To investigate the gut microbial characteristics and risk factors in people with mild cognitive impairment. Methods Twenty-eight people with mild cognitive impairment and 65 age-, gender- as well as education-matched controls were recruited. Carotid artery ultrasonography was conducted and fasting anticoagulant blood and fresh fecal samples were collected on all subjects. Total bacterial DNA was extracted from the stool samples and amplified through real-time quantitative polymerase chain reaction (qPCR). Next generation analysis of 16S rRNA was performed to achieve genomic sequences using the Illumina Hiseq 2500 platform. Results Compared with the control group, mild cognitive impairment group exhibited greater microbial diversity (Bray Curtis distance, Adonis test, P<0.05), which were characterized by enrichment of proteobacteria, Synergistetes, lactobacillus, and depletion of Epulopiscium. Age, creatinine, average CCAIMT, Crouse score, and plaque numbers were associated with cognitive impairment. Multiple logistic regression analysis revealed that the number of hyperechoic plaques (OR=11.511, 95%CI:1.406~94.226, P=0.023) and the abundance of lactobacillus (OR=4.894, 95%CI: 1.090~21.966, P=0.038) were the important risk factors for mild cognitive impairment. Conclusion Higher abundance of lactobacillus in the gut holds potential as novel risk factor of people with mild cognitive impairment.