The human brain-derived neurotrophic factor (hBDNF) gene was cloned by polymerase chain reaction and the recombinant adeno-associated viral vector inserted with hBDNF gene (AAV-hBDNF) was constructed. Cultured rat hippocampal neurons were treated with Abeta(25-35) and serued as the experimental Abeta-induced neuronal damage model (AD model), and the AD model was infected with AAV-hBDNF to explore neuroprotective effects of expression of BDNF. Cell viability was assayed by MTT. The expression of bcl-2 anti-apoptosis protein was detected by immunocytochemical staining. The change of intracellular free Ca ion ([Ca2+]i) was measured by laser scanning confocal microscopy. The results showed that BDNF had protective effects against A-induced neuronal damage. The expression of the bcl-2 anti-apoptosis protein was raised significantly and the balance of [Ca2+]i was maintained in the AAv-hBDNF treatment group as compared with AD model group. These data suggested that recombinant AAV mediated a stable expression of hBDNF in cultured hippocampal neurons and resulted in significant neuron protective effects in AD model. The BDNF may reduce neuron apoptosis through increasing the expression of the bcl-2 anti-apoptosis protein and inhibiting intracellular calcium overload. The viral vector-mediated gene expression of BDNF may pave the way of a novel therapeutic strategy for the treatment of neurodegenerative diseases such as Alzheimer's disease.

The human brain-derived neurotrophic factor (hBDNF) gene was cloned by polymerase chain reaction and the recombinant adeno-associated viral vector inserted with hBDNF gene (AAV-hBDNF) was constructed. Cultured rat hippocampal neurons were treated with Aβ25-35 and serued as the experimental Aβ-induced neuronal damage model (AD model), and the AD model was infected with AAV-hBDNF to explore neuroprotective effects of expression of BDNF. Cell viability was assayed by MTT. The expression of bcl-2 anti-apoptosis protein was detected by immunocytochemical staining. The change of intracellular free Ca ion ([Ca2+]i) was measured by laser scanning confocal microscopy. The results showed that BDNF had protective effects against Aβ-induced neuronal damage. The expression of the bcl-2 anti-apoptosis protein was raised significantly and the balance of [Ca2+]i was maintained in the AAV-hBDNF treatment group as compared with AD model group. These data suggested that recombinant AAV mediated a stable expression of hBDNF in cultured hippocampal neurons and resulted in significant neuron protective effects in AD model. The BDNF may reduce neuron apoptosis through increasing the expression of the bcl-2 anti-apoptosis protein and inhibiting intracellular calcium overload. The viral vector-mediated gene expression of BDNF may pave the way of a novel therapeutic strategy for the treatment of neurodegenerative diseases such as Alzheimer's disease.

Objective To investigate the relationship between hypoxia-related genes and ischemic stroke,and to screen for molecular markers related to hypoxia. Methods First,GSE16561 and GSE58294 datasets and hypoxia-related gene sets were downloaded to screen for hypoxia-related differentially expressed genes. After functional annotation,these hypoxia-related differentially expressed genes were analyzed by the least absolute shrinkage and selection operator least absolute shrinkage and selection operator(LASSO) regression and support vector machine algorithm to screen for immune-related biomarkers,and differential immune cells were analyzed between the ischemic stroke group and the control group. Finally,transcription factors and miRNAs were predicted. Results A total of 13 hypoxia-related differentially expressed genes were screened,which were enriched mainly in the processes of glucose transmembrane transport,monosaccharide transmembrane transport,and mitophagy regulation. Based on the LASSO regression and support vector machine algorithm analyses,eight hypoxia-related biomarkers were identified,which were significantly correlated with most immune cells. Furthermore,the network of 209 transcription factors corresponding to 8 genes and 25 miRNAs corresponding to 6 genes were predicted. Conclusion Eight hypoxia-related biomarkers are identified,which may be used as potential therapeutic targets for inhibiting brain injury after stroke. In addition,glucose metabolism and immune mechanisms may play an important role in the occurrence and progression of ischemic stroke.

Objective To evaluate the optimum compatibility of nabufine mixed with flurbiprofen for patient-controlled intravenous analgesia (PCIA) after gynecological laparoscopic surgery.Methods A total of 210 patients,aged 18-64 yr,with body mass index of 18-30 kg/m2,of American Society of Anesthesiologist physical status Ⅰ or Ⅱ,scheduled for gynecological laparoscopic surgery under general anesthesia,were divided into 4 groups using a random number table method:sufentanil 2.0 μg/kg+flurbiprofen axetil 2.0 mg/kg group (SF group,n =55),nalbuphine 1.5 mg/kg+flurbiprofen axetil 2.0 mg/kg group (N1 F group,n=49),nalbuphine 2.0 mg/kg+flurbiprofen axetil 2.0 mg/kg group (N2F group,n =55) and nalbuphine 3.0 mg/kg +flurbiprofen axetil 2.0 mg/kg group (N3F group,n=51).PCIA solution was prepared correspondingly after surgery in each group.The PCA pump was set up to deliver a 1 ml bolus dose with a 15-min lockout interval and background infusion at 2.0 ml/h.Nalbuphine 5 mg or sufentanil 5 μg was intravenously injected as a rescue analgesic to maintain visual analogue scale score at rest ＜4 at 48 h after surgery in SF and N1 F-N3F groups.Ramsay sedation scores were recorded on admission to post-anesthesia care unit (T1),at the time of post-anesthesia care unit discharge (T2) and at 6,24 and 48 h after surgery (T3-5).The total pressing times of PCIA in 0-6 h,6-24 h and 24-48 h periods after surgery and requirement for rescue analgesics were recorded.The incidence of adverse reactions such as nausea and vomiting,drowsiness and shivering within 48 h after surgery was also recorded.Results Compared with group SF,the incidence of nausea and vomiting was significantly decreased in N1 F and N2F groups,the requirement for rescue analgesics was significantly decreased,and the total pressing times of PCIA was reduced in N2F and N3 F groups,and Ramsay sedation scores at T3,4 were significantly increased in group N3F (P＜0.05).Compared with group N1 F,the requirement for rescue analgesics was significantly decreased,and the total pressing times of PCIA was reduced in N2F and N3F groups,and the incidence of nausea and vomiting and Ramsay sedation scores at T3,4 were significantly increased in group N3F (P＜0.05).Compared with group N2F,the incidence of nausea and vomiting was significantly increased (P＜ 0.05),and no significant change was found in the requirement for rescue analgesics,total pressing times of PCIA or Ramsay sedation scores in group N3F (P＞0.05).Conclusion Nabufine 2.0 mg/kg mixed with flurbiprofen 2.0 mg/kg is the optimum compatibility when used for PCIA after gynecological laparoscopic surgery.

Objective:To summarize the clinical characteristics of adenovirus (HAdVs) pneumonia in children.Methods:The clinical manifestations, laboratory results, imaging features and treatment courses of 541 children with HAdVs pneumonia treated in Shenzhen Children′s Hospital from January 2017 to August 2019 were retrospectively studied.Results:The number of children diagnosed with HAdVs pneumonia increased year by year.A total of 541 patients were included in this study (214 females and 327 males, median age 31 months). The majority of patients (89.3%, 483/541 cases) were less than 6 years of age.Cough (98.3%, 532/541 cases) was the most common symptom, followed by fever (93.7%, 507/541 cases), tachypnea (36.6%, 198/541 cases) and wheeting (32.5%, 176/541 cases). Thirteen point three percent patients (72/541 cases) had existing primary diseases.Severe HAdVs pneumonia was observed in 33.8% of patients (183/541 cases), the most common intrapulmonary complication were pleural effusion(18.6%, 101/541 cases) and respiratory failure (7.0%, 38/541 cases). In co-infection cases, 46.9% of the patients (254/541 cases) suffered from co-infection with mycoplasma pneumoniae, 21.6% of the patients (117/541 cases) suffered from bacterial co-infection.Ninety-eight point five percent of the patients (533/541 cases) were cured, and 1.5% of the patients (8/541 cases) died or discharged voluntarily.Convulsions [ P=0.026, OR=7.312 (95% CI: 1.267-42.191)] and bacterial co-infections [ P=0.030, OR=2.724 (95% CI: 1.101-6.741)] were independently correlated with the occurrence of severe HAdVs pneumonia. Conclusions:Pre-school aged children are at higher risks of developing HAdVs pneumonia.The majority of patients presents with fever and cough as primary manifestations.Tachypnea and wheezing are also common clinical features of HAdVs pneumonia patients.Approximately a third of children with HAdVs pneumonia may develop into severe pneumonia.The presentation of convulsion and the exis-tence of bacterial co-infections may help to predict the development of severe HAdVs pneumonia in pediatric patients.

Objective:To investigate the effects and mechanism of metformin on the wound healing of full-thickness skin defects in diabetic rats.Methods:This study was an experimental study. Eighteen 8-week-old male Sprague Dawley rats were divided into control group, diabetes group, and diabetes+metformin group according to complete random grouping method, with 6 rats in each group. The latter two groups of rats were used to create diabetic models, and then four circular full-thickness skin defect wounds with a diameter of 5 mm were made on the back of 18 rats. Metformin F-127 hydrogel was applied only to the wounds of rats in diabetes+metformin group. The wound healing status on post injury day (POD) 7 and 13 was observed and the wound healing rate was calculated. The wound tissue on POD 7 and 13 was collected for hematoxylin-eosin staining to measure the length of re-epithelialized epidermis and calculate the change rates in diameters of epidermal and dermal wounds, for immunohistochemical staining to detect the relative expressions of keratin 10 and proliferating cell nuclear antigen (PCNA), and for Western blotting to detect the protein expressions of keratin 10 and PCNA. The sample size in all the above experiments was 8 except that in the last experiment was 3. The correlations between the relative expressions of keratin 10 and PCNA in wound tissue in three groups of rats and their wound healing rates, and the correlation between the relative expressions of keratin 10 and PCNA in wound tissue were analyzed.Results:On POD 7, the wound healing rates of rats in diabetes group and diabetes+metformin group were 81.48% (77.89%, 85.53%) and 93.04% (92.51%, 94.24%), which were significantly lower than 100% (97.17%, 100%) in control group (with Z values of 2.37 and -3.36, respectively, P<0.05); the wound healing rate of rats in diabetes+metformin group was significantly higher than that in diabetes group ( Z=3.45, P<0.05). On POD 13, the wound healing rates of rats in control group and diabetes+metformin group were both 100% (100%, 100%), which were significantly higher than 94.47% (90.68%, 99.82%) in diabetes group (with Z values of 2.90 and -2.90, respectively, P<0.05). On POD 7, the change rates in epidermal wound diameter of rats in control group and diabetes+metformin group were significantly higher than that in diabetes group (with Z values of 3.36 and -2.74, respectively, P<0.05). The change rates in dermal wound diameter of rats in the three groups were similar on POD 7 and 13 ( P>0.05). The lengths of re-epithelialized epidermis of rats in control group and diabetes+metformin group on POD 13 were significantly longer than that in diabetes group (with Z values of 3.34 and -2.64, respectively, P<0.05). The relative expressions of keratin 10 in wound tissue of rats in diabetes group on POD 7 and 13 were significantly higher than those in control group (with Z values of -3.36 and -3.26, respectively, P<0.05) and diabetes+metformin group (with Z values of 3.36 and 3.15, respectively, P<0.05), and the relative expression of keratin 10 in wound tissue of rats in diabetes+metformin group on POD 7 was significantly lower than that in control group ( Z=3.05, P<0.05); the relative expressions of PCNA in wound tissue of rats in diabetes group on POD 7 and 13 were significantly lower than those in control group (with both Z values of 3.36, P<0.05) and diabetes+metformin group (with both Z values of -3.36, P<0.05). The protein expressions of keratin 10 in wound tissue of rats in control group and diabetes+metformin group on POD 7 as well as that in diabetes+metformin group on POD 13 were significantly lower than those in diabetes group ( P<0.05), and the protein expressions of PCNA in wound tissue of rats in control group and diabetes+metformin group on POD 7 were significantly higher than that in diabetes group ( P<0.05). There was a significant positive correlation between the relative expression of keratin 10 in wound tissue and the wound healing rate in control group and diabetes+metformin group of rats (with r values of 0.78 and 0.71, respectively, P<0.05), there was a significant negative correlation between the relative expression of PCNA in wound tissue and the wound healing rate in diabetes+metformin group of rats ( r=-0.60, P<0.05), and there was a significant negative correlation between the relative expressions of PCNA and keratin 10 in wound tissue of rats in diabetes group and diabetes+metformin group (with r values of -0.41 and -0.49, respectively, P<0.05). Conclusions:The diabetic rats with full-thickness skin defect wound exhibit delayed healing, accompanied by up-regulation of keratin 10 and down-regulation of PCNA in keratinocytes in the wound tissue. Metformin can promote wound healing in diabetic rats with full-thickness skin defects by down-regulating keratin 10 expression and up-regulating PCNA expression in keratinocytes in the wound tissue, and the wound healing rate was positively correlated with the expression of keratin 10 and negatively correlated with the expression of PCNA.

Objective:To investigate the surgical effect of various circulatory pathways on replantation of severed distal segments of digits.Methods:From June 2017 to June 2023, a total of 137 patients (162 digits) who received digit replantation surgery in the Department of Trauma Orthopaedics and Microsurgery of Guilin People's Hospital were retrospectively studied. The patients were 82 males (97 digits) and 55 females (65 digits) aged 11.0 months to 51.0 years old, with 31.7 years old in average. The injured digits were 36 thumbs, 43 index fingers, 32 middle fingers, 28 ring fingers and 23 little fingers. Causes of injury: 55 digits were of electric saw, 68 of sharp object cut, 26 of twisting crush and 13 of degloving injury. According to the circulatory pathways, 66 patients (88 digits) were assigned to a physiological replantation group (PRG), 50 patients (53 digits) to an arterio-venolisation group (AVG) and 21 patients (21 digits) to a nail bed bloodletting group (NBG). Functional recovery was evaluated according to the Evaluation Standard of Severed Finger Functional of Hand Surgery of Chinese Medical Association. Analysis of variance or chi-square test was employed to analyse the age, gender, time from injury to surgery, surgery time, incidence of vascular compromise of the replanted digit, survival rate, score of functional recovery of the affected digit at 6 months after surgery, and incidence of postoperative complications in each group. P<0.05 was considered statistically significant. Results:All patients were included in the 6-10 months of postoperative follow-up, with an average of 7.2 months. Regarding the score of functional recovery of affected digit according to the Evaluation Standard of Severed Finger Functional of Hand Surgery of Chinese Medical Association, PRG (86.3 points±7.9 points) was higher than that of NBG (78.3 points±11.2 points) and AVG (70.7 points±8.6 points), with statistically significant differences ( P<0.05 for both). In terms of the incidence of vascular compromise, PRG (3 digits for 3.4%) was lower than that of NBG (1 digit for 4.8%) and AVG (6 digits for 11.3%), with statistically significant differences ( P<0.05 for both). Of the survival rate, PRG (81 digits for 92.0%) was superior to that of NBG (18 digits for 85.7%) and AVG (42 digits 79.2%), with statistically significant differences ( P<0.05 for both). While of the incidence of postoperative complications, PRG (12.6%) was lower than that of AVG (17.1%) and NBG (21.2%), with statistically significant differences ( P<0.05 for both). Conclusion:For a severed distal segment of digit, the physiological replantation is preferred. For the digit with undetectable return veins, effective replantation can be achieved by the arterio-venolisation or nail bed exsanguination.

Objective:To explore the application value of whole-process ultrasound-guided percutaneous portal vein puncture islet transplantation.Methods:From October 2018 to May 2021, 16 diabetics underwent whole-process ultrasound-guided percutaneous portal vein puncture islet transplantation at First Affiliated Hospital of Sun Yat-sen University.The whole process was guided by ultrasound for completing percutaneous portal vein puncture catheterization, islet infusion monitoring, bleeding prevention and ablation hemostasis after bleeding.Results:Ten patients [8 males and 2 females with a mean age of(45.9±21.1)years]underwent 16 islet transplants, including one islet(5 cases), two islets(4 cases)and three islets(1 case). A single puncture was successfully performed without damage to other extrahepatic organs, persistent portal hypertension, portal vein embolism or infection.Bleeding at liver puncture site occurred in 3 cases and ultrasound radiofrequency ablation was performed for immediate hemostasis.Among them, postoperative blood glucose stabilized at 4~12 mmol/l post-operation.And 5 cases(31.3%)achieved insulin independence for>2 months and 10 cases(62.5%)lowered insulin dosage by>50% as compared with preoperative level.The level of fasting C-peptide recovered or was higher than normal in 10 cases(62.5%)and became obviously elevated in the remainders.In 11 cases(68.8%)of them, liver transaminase was briefly and mildly elevated post-operation, and no other complications were observed.Conclusions:The whole-process ultrasound-guided percutaneous portal vein islet transplantation is both safe and feseasible.It avoids the injury of transplanted kidney caused by contrast agent and radiological radiation to operator and patient.It is a method of islet transplantation worth a wider popularization.

BACKGROUND:Increasing evidence suggests that N6-methyladenosine(m6A)regulators are closely associated with osteoarthritis and are considered to be a new direction in the prevention and treatment of osteoarthritis,but their specific mechanism of action is unknown. OBJECTIVE:To conduct a bioinformatics analysis of the osteoarthritis gene microarray dataset in order to explore the role of m6A in osteoarthritis and analyze the pathogenesis of osteoarthritis. METHODS:The m6A regulators associated with osteoarthritis and their expression were first extracted from the GSE1919 dataset in the GEO database using R software,and then the results were analyzed by gene difference analysis and GO and KEGG enrichment analyses.Subsequently,the results of protein-protein interaction network topology analysis and machine learning results were intersected to obtain the m6A Hub regulators,which were validated by in vitro cellular experiments. RESULTS AND CONCLUSION:A total of 16 osteoarthritis-related m6A regulators were extracted and 11 m6A differential regulators,including ZC3H13,YTHDC1,YTHDF3 and HNRNPC,were obtained by differential analysis.GO enrichment analysis showed that osteoarthritis-related m6A differential regulators played a role in the biological processes such as mRNA transport,RNA catabolism,and regulation of insulin-like growth factor receptor signaling pathway.(3)KEGG enrichment analysis showed that the differential regulators were mainly involved in the p53,interleukin-17 and AMPK signaling pathways.The combined protein-protein interaction network topology analysis and machine learning results obtained the m6A Hub regulator-YTHDC1.(5)The results of in vitro cellular experiments showed that there was a significant difference in the expression of m6A key regulator between the control and experimental groups(P<0.05).To conclude,YTHDC1 is closely related to the development of osteoarthritis,which is expected to be a molecular target of m6A for the treatment of osteoarthritis.

Objective:To investigate the impact of BMI1 expression in OSCC on the recruitment and differentiation of tumor-associat-ed macrophages(TAMs).Methods:BMI1 expression in 519 cases of OSCC tissues and 44 normal controls was analyzed using online datasets of GEPIA 2.0,and validated in 3 cases of OSCC samples and controls by qRT-PCR and western blotting.The function of BMI1/NF-κB axis during OSCC carcinogenesis was investigated by CCK8 assays,wound healing test and transwell assays.Macrophage phenotypes and recruitment were determined using qRT-PCR and western blotting following coculture of the cells with human monocyte cells(THP-1)by OSCC conditioned medium.Moreover,a cell line-derived xenograft(CDX)model was used to detect the effect of BMI1 on tumor growth in vivo.Results:Compared with the normal tissues and cells,the expression level of BMI1 in OSCC tissues and cells was significantly upregulated.BMI1 knockdown impaired the proliferation,migration,and invasion abilities of OSCC cell lines in NF-κB-dependent manner.Furthermore,OSCC cells with high BMI1 expression inhibited the migration of THP-1 cells,promoted M2-like macrophage polarization through NF-κB pathway in vitro.Xenograft experiments further confirmed the inhibitory effect of BMI1 knockdown on the tumorigenesis ability of OSCC cells in vivo.Conclusion:BMI1 promotes M2-like polarization by regulating NF-κB and may be used as a potential therapeutic target for antitumor immunity.