Objective:To investigate the knowledge of Sixth Chinese national consensus report on the management of Helicobacter pylori infection ( treatment excluded) (hereinafter referred to as sixth national consensus) and 2022 Chinese national clinical practice guideline on Helicobacter pylori eradication treatment (hereinafter referred to as the guideline)among medical staff from general hospitals in Hainan. Methods:From February 20 to May 7, 2023, a questionnaire survey on the diagnosis and treatment of Helicobacter pylori ( H. pylori) infection was conducted among 1 463 medical staff from 15 general hospitals in Hainan Province. The questionnaire was drawn up according to the sixth national consensus and the guideline, covering knowledge of 6 sections, induding H. pylori related diseases, detection of H. pylori, eradication, prevention and influence factors of eradication of H. pylori, etc. Chi-square test was used for statistical analysis. Results:A total of 1 463 valid questionnaires were collected with the effective responsive rate of 100.00%.The 1 463 subjects included 225 gastroenterologists and 1 238 other medical staff(including 503 physicians from other departments, 264 surgeons and 471 medical technologists and pharmacists). About 78.67%(177/225)of gastroenterologists agreed that the overall infection rate of H. pylori in China was more than 20%, the awareness rate was higher than that of other medical staff (physicians from other departments 65.41%(329/503), surgeons 61.74%(163/264), medical technologists and pharmacists 60.30%(284/471); the following datas were sorted by this position), and the difference was statistically significant ( χ2=30.97, P<0.001). About 51.11%(115/225) of gastroenterologists considered that H. pylori serological antibody test could not be used as a diagnostic method for current infection, the awareness rate was higher than that of other medical staff(22.07%(111/503), 14.02%(37/264), 12.31%(58/471)), and the difference was statistically significant( χ2 =152.66, P<0.001). Proton pump inhibitor and potassium-competitive acid blocker should be discontinued for 2 weeks, and antibiotics and bismuth should be discontinued for 4 weeks before urea breath test, and the awareness rates of gastroenterologists were higher than those of other medical staff (38.67%(87/225) vs. 23.26%(117/503), 19.70%(52/264), 18.47%(87/471); 60.89%(137/225) vs. 26.64%(134/503), 25.76%(68/264), 23.78%(112/471)), and the differences were statistically significant ( χ2 =133.70 and 165.51, both P<0.001). For refractory H. pylori infection, 98.67%(222/225)of gastroenterologists agreed with the individualized diagnosis and treatment of H. pylori infection should be guided by bacterial culture, antibiotic susceptibility test or drug resistance gene test, and the awareness rate was higher than that of other medical staff (91.85%(462/503), 93.56%(247/264), 93.21%(439/471)), and the difference was statistically significant( χ2=20.55, P=0.002). About 70.67% (159/225) of gastroenterologists recommended a bismuth containing quadruple regimen, 80.44% (181/225) supported a 10 to 14 day H. pylori eradication course, and the awareness rates were higher than other medical staff (46.92%(236/503), 33.33%(88/264), 32.91%(155/471); 67.20%(338/503), 59.09%(156/264), 53.93%(254/471)), and the differences were statistically significant ( χ2=111.25 and 59.99, both P<0.001). The understanding rates of the sixth national consensus and the guideline in gastroenterologists was 85.33% (192/225), which was higher than that of other medical staff (64.21%(323/503), 66.67%(176/264), 57.96%(273/471)), and the difference was statistically significant ( χ2=85.47, P<0.001). Conclusions:Gastroenterologists from general hospitals in Hainan Province have a better understanding of the sixth national consensus and the guideline than other medical staff. However, there is still a lack of deep understanding of the sixth national consensus and the guideline, and it is necessary to further strengthen the learning and application of the sixth national consensus and the guideline.

Objective Aiming at improving biomechanical strength of the anastomotic stoma as well as reducing tissue thermal damage, a novel radiofrequency (RF) tissue welding electrode was developed. Methods A novel electrode with a hollow structure on the surface ( the plum electrode) was designed and the ring electrode was used as control group to conduct the welding of intestinal tissues based on RF energy. Biomechanical properties of anastomotic stoma were studied by shear test and burst pressure test. The tissue thermal damage during welding was investigated by finite element electro-thermal-mechanical multi-field coupling simulation analysis and thermocouple probe, and the tissue microstructures were also studied. Results Under 120 W RF energy, 8 s welding duration and 20 kPa compression pressure, the anastomotic stoma had the optimal biomechanical properties. Compared with the ring electrode group, biomechanical strength of the anastomotic stoma in plum electrode group was higher, with the shear strength and burst pressure increasing from (9. 7±1. 47) N, (84. 0±5. 99) mmHg to (11. 1±1. 71) N, (89. 4±6. 60) mmHg, respectively. There was a significant reduction in tissue thermal damage, and intact and fully fused stomas could be formed in anastomotic area. Conclusions The proposed novel electrode could improve biomechanical strength of the anastomosis as well as reduce tissue hermal damage, thus achieve better fusion. The research result provide references for realizing the seamless connection of human lumen tissues

Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.

Objective: To compare the osteogenic differentiation abilities of human bone marrow mesenchymal stem cells (hBMSCs) from different sources, and to provide basis for choosing a new source of seed cells in bone tissue engineering. Methods: Jaw bone-marrow-derived mesenchymal stem cells (JMMSCs) were isolated from orthognathic surgical sites and cultured by limited dilution for single cell clone. Long bone-marrow-derived mesenchymal stem cells (BMMSCs) were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method. Flow cytometry was used to detect the surface markers of both cells. Osteogenic ability was assessed by PCR and Western Blot after osteogenic differentiation for the following molecules: Runx2, COL-1 and OCN. Alizarin red staining was used for determining the ability of cell mineralization after osteogenic differentiation. Results : The expressions of cell surface markers CD90 and CD105 were positive in both type of cells, while CD34, CD14 and CD45 were all negative. After 21 days of osteogenic induction, JMMSCs formed significantly more mineralized nodules than BMMSCs. After 7, 14, 21 days of osteogenic induction, JMMSCs expressed more osteogenic-related molecules than BMMSCs. Conclusion The osteogenic differentiation capacity and mineralization ability of JMMSCs are significantly higher than BMMSCs. Jaw bone might be a more suitable source of seed cells in bone tissue engineering compared with long bone.

Objective:To compare acetyltransferase MORF level in periodontal ligament stem cells( PDLSCs) derived from healthy individuals ( H-PDLSCs) with those derived from the individuals with periodontitis ( P-PDLSCs ) . And to determine the effect of MORF on the osteogenic differentiation potential of PDLSCs. Methods: Human H-PDLSCs and P-PDLSCs were cultured and cloned with limited dilution method. H-PDLSCs were stimulated by LPS, TNF-α, IL-β and the mix of the 3 inflammatory factors to imitate inflammatory environment ( IP-PDLSCs ) . Quantitative RT-PCR and Western Blot were applied to examine different expression of MORF in H-PDLSCs and P-PDLSCs. Western Blot was applied to detect expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red staining were applied to determine osteogenic differentiation potential of H-PDLSCs with MORF knock-down. Results:Quantitative RT-PCR and Western Blot showed lower expression of MORF in P-PDLSCs compared with H-PDLSCs( P<0. 05). Western Blot revealed lower expression of MORF in IP-PDLSCs. Quantitative RT-PCR, Western Blot and alizarin red stai-ning indicated osteogenic differentiation potential was inhibited in H-PDLSCs with MORF knockdown(P<0. 05). Conclusion: Peri-odontitis can suppress the expression of MORF in PDLSCs and inhibite the osteogenic differentiation potential of PDLSCs.

Objective:To compare the proliferation and osteogenic differentiation between human bone marrow mesenchymal stem cells from orofacial bone(OMMSCs)and those from long bone(BMMSCs).Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.BMMSCs were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method.The surface markers of the cells were detected by flowcytometry.Single-colony formation,CCK assay and cell circle analyses were conducted.Osteogenic differentiation ability was evaluated by ALP activity test and Alizarin red staining after osteogenic induction culture.Results:The cell surface markers STRO-1 and CD105 of both stem cells were positive,CD34,CD31 and CD45 were negative. OMMSCs generated significantly higher numbers of colonies than BMMSCs.In addition,OMMSCs had a higher proliferation rate and more cells in proliferative(S +G2 )stage than BMMSCs.After osteogenic induction for 3,5,7 and 10 d,OMMSCs showed higher levels of ALP activity.OMMSCs formed significantly more mineralized nodules than BMMSCs after 21-day ostogenic induction.Conclusion:The proliferation and osteogenic differentiation capacity of OMMSCs are higher than those of BMMSCs.

Aim To evaluate the effects of morphine preconditioning ( MPC ) on the expression of microR-NAs ( miRNAs ) induced by hypoxia-reoxygenation (H/R) in H9c2 myocardial cells. Methods H9c2 cells were randomly divided into 3 groups ( n=4 each) as follows:control group ( CON) , hypoxia/ reoxygen-ation group ( H/R ) and morphine preconditioning group ( MPC+H/R) . The cells were cultured in nor-mal condition in CON group. The cells were subjected to 5 h hypoxia followed by 1 h reoxygenation in H/R group and MPC+H/R group. Specifically, the cells in MPC+H/R group were preconditioned with morphine with the final concentration of 1 μmol·L-1 for 10 min before H/R. After the treatment, CCK-8 was used to detect cell viability and chemical colorimetry was used to detect lactate dehydrogenase ( LDH ) activity in the culture medium. Cell apoptosis was assessed by An-nexin-V-FITC/PI flow cytometry. Relative expression of Fas protein was detected by Western blot. The ex-pression of miRNA in myocardial cells was analyzed by quantitative reverse transcription polymerase chain re-action ( qRT-PCR ) . Results Compared with CON group, the cell viability was significantly decreased, while the LDH activity, apoptotic rate and Fas protein expression were dramatically increased in group H/R (P<0. 01). However, MPC significantly increased the cell viability, whereas it decreased the LDH activity, apoptotic rate and Fas protein expression induced by H/R injury ( P < 0. 01 ) . The expressions of miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7 e-5 p were markedly down-regulated by H/R as compared to CON group ( P <0. 05 ) , while MPC inhibited these miRNAs which were significantly down-regulated by H/R group ( P <0. 01 ) . Conclusion Morphine preconditioning might protect H9 c2 myocar-dial cells against H/R injury by regulating the expres-sion of miRNAs such as miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7e-5p.

Objective To evaluate the role of microRNA-133b-Sp (miR-133b-Sp) in apoptosis hypoxia/reoxygenation (H/R) induced by in rat cardiomyocytes.Methods Rat myocardial cell line H9c2 was cultured in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum.The cells were seeded in 96-well or 6-well plates and randomly divided into 4 groups (n=64 wells each):control group (group C);group H/R;miR-133b-5p mimic + H/R group (group M+H/R);miR-133b-Sp negative control + H/R group (group NC+H/R).The cells were exposed to 95％ N2-5％ CO2for 5 h at 37 ℃ followed by 1 h reoxygenation in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum in all the groups except group C.The cells were cultured in normal culture atmosphere in group C.In M+H/R and NC+H/R groups,the cells were transfected with miR-133b-5p mimic (final concentration 30 nmol/L) and miR-133b-5p negative control (final concentration 30 nmol/L),respectively,for 24 h before H/R.Total RNA was extracted from cells to detect the expression of miR-133b-5p using quantitative real-time PCR.The cell viability (by CCK-8) and lactic dehydrogenase (LDH) activity in the culture medium were detected.Cell apoptosis was assessed by Annexin V/PI flow cytometry.Apoptotic rate was calculated.Result Compared with group C,the cell viability was significantly decreased,and the LDH activity and apoptotic rate were increased in H/R,M+H/R and NC+H/R groups,the expression of miR-133b-5p was down-regulated in H/R and NC+H/R groups,and the expression of miR-133b-Sp was up-regulated in group M+H/R.Compared with group H/R,the cell viability was significanttly increased,the LDH activity and apoptotic rate were decreased,and the expression of miR-133b-5p was up-regulated in group M+H/R,and no significant change was found in the parameters mentioned above in group NC+H/R.Conclusion H/R in rat cardiomyocytes can induce cell apoptosis possibility through down-regulating the expression of miR-133b-5p

Objective To evaluate the effect of morphine preconditioning on the expression of miR-133b-Sp and Fas in rat cardiomyocytes subjected to hypoxia/reoxygenation (H/R).Methods Cardiomyocytes were isolated from healthy adult male Sprague-Dawley rats by using Langendorff perfusion.The cells were seeded into 24-well plates or 60 mm diameter dishes and randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),group H/R,and morphine preconditioning group (group MPC).The cells in group C were cultured in normal culture atmosphere.In H/R and MPC groups,the cells were exposed to 95％ N2-5％ CO2 for 90 min followed by 120 min reoxygenation.In group MPC,the cells were cultured for 10 min in serum-free DMEM liquid culture medium containing morphine 1 μmol/L,and then were cultured for 30 min in morphine-free DMEM liquid culture medium before hypoxia.At 120 min of reoxygenation,the cells in 24-well plates were selected to detect the cell viability (by MTT),lactate dehydrogenase (LDH) activity in the culture medium,and cell apoptosis (by Hoechst 33234 staining).Apoptosis rate was calculated.Total RNA and protein were extracted from the cells in 60 mm dishes to detect the expression of miR-133b-5p and Fas mRNA (by quantitative real-time PCR) and Fas protein (by Western blot).Results Compared with C group,the cell viability was significantly decreased,LDH activity and apoptosis rate were increased,the expression of miR-133b-Sp was down-regulated,and the expression of Fas mRNA and protein was up-regulated in H/R group.Compared with H/R group,the cell viability was significantly increased,LDH activity and apoptosis rate were decreased,the expression of miR-133b-5p was up-regulatcd,and the expression of Fas mRNA and protein was down-regulated in MPC group.Conclusion The mechanism by which morphine preconditioning reduces H/R injury to rat cardiomyocytesis related to up-regulation of the expression of miR-133b-Sp and down-regulation of the expression of Fas.

Objective To study the clinical features and angiographic findings of moyamoya disease (MMD) as well as their relationship. Methods A total of 22 MMD patients received routine digital substraction angiography (DSA). The clinical manifestations and angiographic findings were analyzed. Results Clinical manifestations varied and each patient often had multiple symptoms, including cerebral infarction in 9 patients with an average age of 23.6 (13-39 years) and cerebral hemorrhage in 7 patients with an average age of 31.2 (28-46 years). Angiographic examination found that all the diseased sides showed MMD blood vessels. The patients who received encephalo-myo-arterio-synangiosis (EMAS) had better prognosis than those without receiving the treatment. Conclusion Cerebral infarction is frequent in children and adolescents with MMD, whereas cerebral hemorrhage is common in adults. DSA is a golden criterion for diagnosing MMD. Surgical treatment, EMAS blood supply reconstruction in particular, should be prescribed.