Diabetic retinopathy(DR), the most common ocular complication of diabetes, is one of the major causes of vision impairment and even blindness among the working population as well as middle-aged and elderly individuals. In the diabetic microvascular system, hyperglycemia damages retinal endothelial cells, enhances vascular permeability and disrupts the blood-retinal barrier through different mechanisms, all of which result in endothelial dysfunction. Retinal vascular dysfunction caused by multifactors, such as peroxisome proliferator-activated receptor-y disruption, oxidative stress, inflammation, increased advanced glycation end products and their receptors, and microRNA dysregulation can cause vascular endothelial damage, accelerate retinal endothelial dysfunction, lead to the progression of DR. Therefore, the available date and the contributors in the pathophysiology of DR are reviewed with a special emphasis on the retinal endothelial dysfunction, for a better understanding of the molecular cellular mechanism in the development of DR, to analyze the challenges in the treatment of DR and to provide new ideas and strategies for the clinical management and treatment of DR.

Objective:To analyze the F10 gene mutations in a Chinese pedigree affected with the deficiency of the hereditary coagulation factor X (FX), resulting from a new deletion mutation, and to study the associated molecular pathogenesis.Methods:Next generation sequencing (NGS) was performed to screen the genetic mutations in the proband which were then verified by Sanger sequencing. The FX activity (FX∶C) of probands and their family members was detected using the blood clotting method, and the mutation sites of the family members were analyzed using Sanger sequencing. The pathogenicity of the mutation site was predicted by using the online bioinformatics software, Mutation Taster. The SWISS-MODEL software was used for stimulating the three-dimensional models of the wild-type and mutant proteins for analyzing the influence of the mutation site on the structure and function of the proteins, and for analyzing the difference between the catalytic residues of the wild-type and the mutant proteins. The level of the F10 gene mRNA was quantitatively analyzed by qRT-PCR (quantitative reverse transcription polymerase chain reaction) method by constructing plasmids, transfecting human embryonic kidney 293T cells (HEK 293T), and analyzing the splicing of the mutated site by RT-PCR method. The levels of FⅩ∶Ag in cell lysates and cell culture media (both inside and outside the cells) were detected by the ELISA (enzyme linked immunosorbent assay) method.Results:A medium-grade factor X deficiency with a 36.42% FⅩ∶C ratio was detected in the proband by the coagulation method. NGS analysis demonstrated a heterozygous deletion mutation in exon 8:c.902_919del (p.Ala301_Glu306del) in the proband. Sanger sequencing analysis indicated that some members of the family (mother and grandfather) were also carriers of the corresponding deletion mutation. Online bioinformatics software predicted the pathogenic nature of the c.902_919del mutation, with a pathogenic score of 0.999. The 3D protein structure model analysis indicated that the c.902_919del mutation resulted in the disappearance of a segment of β-fold in the protein structure, thereby shortening the preceding segment of the β-fold and a subsequent loss of hydrogen bonds between adjacent amino acids with no significant difference in the side chain conformation of the key catalytic residues compared to the wild-type. mRNA splicing analysis indicated the absence of alternative splicing changes in the mutation, and qRT-PCR results indicated the absence of a statistically significant difference between the mRNA levels of F10 gene and wild-type mRNA in cells expressing c.902_919del mutant. The ELISA results indicated that there was no statistically significant difference in the FX∶Ag levels of the mutant cell culture medium and the lysate.Conclusions:In this pedigree, the heterozygous mutation in exon 8 of F10 gene (c.902_919del, p.Ala301_Glu306del) caused the hereditary factor Ⅹ deficiency.

ObjectiveTo compare the efficacy and safety of acupuncture combined with auricular acupoints patches and nicotine transdermal patch in treating moderate to severe nicotine dependence. MethodsIn a rando-mized controlled trial， 64 quit smoking voluntary subjects with moderate to severe nicotine dependence were randomly divided at a ratio of 1∶1 into a treatment group and a control group， with 32 cases in each group. The treatment group was given acupuncture combined with auricular acupoints patches， twice weekly， four weeks as a course for two courses. The control group was given nicotine transdermal patch， one patch per day for 24 hours， 8 weeks. The cure rate was assessed after treatment and at follow-up （the 16th week after treatment）.The daily smoking volume， exhaled carbon monoxide （CO） value， Nicotine Dependence Scale （FTND）， Minnesota Nicotine Withdrawal Symptoms Scale （MNWS）， and Pittsburgh Sleepiness Index Inventory （PSQI） were evaluated before and after treatment and at follow-up， and adverse effects were recorded. ResultsIn terms of the cure rate， there were both six cured cases （20%） after treatment and at follow-up in the treatment group， while in the control group， seven （23.3%） and five （16.7%） patients were cured after treatment and at follow-up， respectively， with no statistically significant differences between the two groups both after treatment and at follow-up （P＞0.05）. The daily smoking volume and exhaled CO value significantly decreased after treatment and at follow-up in both groups （P＜0.05）， but were not significantly different between the groups after treatment and at follow-up （P＞0.05）. After treatment and at follow-up， FTND， MNWS， and PSQI scores were significantly reduced in both groups compared with those before treatment （P＜0.05）. There was no statistically significant difference in the FTND scale scores between the two groups after treatment and at follow-up （P＞0.05）， while the MNWS and PSQI scale scores were lower in the treatment group than in the control group （P＜0.05）. ConclusionAcupuncture combined with auricular acupoint patches for moderate to severe nicotine dependence has comparable effect with the first-line drug nicotine patch in terms of increasing the cure rate and decreasing the degree of nicotine dependence， and is superior to nicotine patch in terms of relieving withdrawal symptoms and improving sleep， with stable long-term effect.

Objective:To explore the effects of multidisciplinary pain management in patients with cesarean scar pregnancy (CSP) undergoing uterine artery embolization (UAE) .Methods:Totally 140 CSP patients who underwent UAE in Nantong Maternity and Child Health Care Hospital, Nantong University from January 2019 to March 2021 were selected by convenient sampling and divided into the observation group and control group according to the random number table, with 70 patients in each group. Patients in the control group received routine care, while patients in the observation group underwent multidisciplinary pain management. The stress response-related indicators, complication rates, and pain control satisfaction were compared between the two groups.Results:The heart rate, systolic blood pressure, visual analog pain score, and anxiety score of the observation group were lower than those of the control group, and the Brinell comfort score was higher than that of the control group, with statistically significant differences ( P<0.05) . The total incidence of complications in the observation group was 8.57% (6/70) , which was lower than 21.43% (15/70) in the control group, and the difference was statistically significant ( P<0.05) . The total satisfaction with pain control in the observation group was 91.43% (64/70) , which was higher than 75.71% (53/70) in the control group, and the difference was statistically significant ( P<0.05) . Conclusions:The multidisciplinary pain management is beneficial to relieve the degree of stress response of patients, reduce the occurrence of complications, and improve the satisfaction with pain control.

Objective:To evaluate the modified efficacy of thoracic paravertebral block (TPVB) combined with general anesthesia in the patients undergoing laparoscopic radical nephrectomy.Methods:Eighty patients, aged 38-64 yr, with body mass index of 18-24 kg/m 2, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective laparoscopic radical nephrectomy, were selected and randomly divided into 2 groups ( n=40 each) using a random number table method: general anesthesia group (group GA) and TPVB combined with general anesthesia group (group TPVB+ GA). A paravertebral catheter was placed at T 8 and T 10 under ultrasound guidance before induction of anesthesia, and 0.5% ropivacaine 10 ml was administered via the catheter in group TPVB+ GA.Anesthesia was induced with propofol, sufentanil, etomidate and rocuronium and maintained by intravenous infusion of propofol and remifentanil.Patient-controlled intravenous analgesia was performed with sufentanil, ketorolac tromethamine and tropisetron at the end of surgery.When postoperative visual analog scale score≥4, tramadol 50 mg was intravenously injected as rescue analgesic.Immediately before anesthesia induction (T 0), at 5 min after establishing pneumoperitoneum (T 1), at 2 h of pneumoperitoneum (T 2), and immediately after the end of pneumoperitoneum (T 3), and at 24 h after operation (T 4), venous blood samples were collected for determination of plasma norepinephrine concentrations (by enzyme-linked immunosorbent assay), plasma cortisol level (using radioimmunoassay), and blood glucose concentrations were measured.The intraoperative consumption of sufentanil and remifentanil was recorded.The intraoperative hypertension, hypotension, and bradycardia were recorded, and the nausea and vomiting, pruritus, and requirement for rescue analgesia occurred within 24 h after surgery were recorded. Results:Compared with group GA, the plasma concentrations of norepinephrine, cortisol and blood glucose were significantly decreased at T 1-4, the intraoperative consumption of sufentanil and remifentanil was reduced, and the postoperative requirement for rescue analgesia was decreased in group TPVB+ GA ( P<0.05). There was no significant difference in the incidence of intraoperative and postoperative adverse reactions between the two groups ( P>0.05). Conclusion:TPVB combined with general anesthesia is helpful in carrying out the anesthetic model of low-consumption opioids and is more helpful in inhibiting intraoperative and postoperative stress responses and postoperative pain responses than general anesthesia alone when used for laparoscopic radical nephrectomy.

OBJECTIVE：To struc turally modify shikimic acid ，and to investigate the reversal effects of its derivatives on paclitaxel-resistant human breast cancer cells MCF- 7/PTX. METHODS ：Using shikimic acid as the lead structure ，1-position carboxyl group was structurally modified to synthesize a series of shikimic acid derivatives through esterification ，amidation， hydrogenation and reduction ，etc. Using non-drug resistant cells MCF- 7 as reference ，MTT assay was used to screen derivatives with inhibitory activity as well as half-inhibitory concentration （IC50）and reversal index （RI）of derivatives to MCF- 7/PTX. With the drug resistance-related transgelin 2 as the target ，the molecular docking of the active derivatives with the drug resistance-related protein was carried out by using Glide 1.0 computer-aided design software. RESULTS ：Totally 15 derivatives were obtained （T1-T15）， of which T 4-T15 were obtained for the first time. MTT assay showed that （3R， 4S， 5R） -N-benzyl-3， 4， 5-trihydroxy-1-cyclohexene-1-formamide（T7），（3R，4S，5R）-N-（3，4，5-trihydroxy-1-cyclohexenylmethyl）-benzylamine（T14）， （3R，4S，5R）-3，4-O-isopropyl-5-O-acetyl-1-cyclohexene-1-methyl formate （T15）inhibited MCF- 7 and MCF- 7/PTX cells to a certain extent ；IC50 values of T 7，T14 and T 15 combined with pacliaxel to MCF- 7/PTX cells were significantly lower than that in negative control （Paclitaxel alone ）group（P＜0.05）. RIs of T 14 and T 15 were higher ，and RIs of the highest dose were 8.8 and 9.3， which were equivalent to positive control verapamil （10.8）. Th e results of molecular docking showed that the hydroxyl groups at positions 3，4 of T 7 could form multiple hydrogen bonds with ； Arg625 and Asp 627 in the catalytic region of transgelin 2. In addition to the hydrogen bond mentioned above at T 7，the mail：batistuta28@126.com secondary amine side chain at position 1 of T 14 could also form hydrogen bond with Glu 657 of transgelin 2. When the hydroxyl group on the T 15 mother nucleus was derived from the donor group ，the binding of the hydroxyl group to transgelin 2 was closer and the inhibition was enhanced. CONCLUSIONS ： The derivatives T 7，T14 and T 15 have certain reverse activity to paclitaxel-resistant human breast cancer cells. The polyhydroxy structure of the mother nucleus is the main structural region of the hydrogen bond between shikimic acid and its derivatives and transgelin 2. The derivation of its power supply group or the introduction of secondary amines and hydrophobic groups into the 1-carboxyl group of shikimic acid is benifit for enhancing the drug resistance reversal effect of derivative .

Objective: To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism. Methods: Human LEC lines (SRA01/04) were divided into MeCP2-mimic group, MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group, and MeCP2 analog plasmid, blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection, the migration rate of the cells was evaluated by scratching test, and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin, E-cadherin, Vimentin, matrix metallo proteinase (MMP)-9, MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot. Results: After 24 hours of transfection, the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group (F=4 773.00, P<0.00 1). The migrating rate of the cells in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was (57.45±5.20)%, (32.71±10.02)% and (17.77±9.22)%, respectively, showing a significant difference among the three groups (F=124.00, P<0.001), and the migrating rate of the cells in the si-MeCP2 group was significantly lower than that of the MeCP2-mimic group or MeCP2-NC group (both at P<0.001). The relative expressing intensity (absorbance) of Wnt3a in the cells of the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 75.92±6.10, 52.03±5.22 and 28.75±3.39, respectively, with a significant difference among three the groups (F=221.30, P<0.001), and the relative expressing intensity (absorbance) of Wnt3a in the cells was significantly lower in the si-MeCP2-mimic group than that of the MeCP2-NC group and MeCP2-mimic group (both at P<0.001). The relative expressing level of E-cadherin protein was significantly elevated and the expressions of β-catenin, Vimentin, MMP-9 and MMP-7 were significantly reduced in the si-MeCP2 group compared with the MeCP2-mimnic group and MeCP2-NC group (all at P<0.01). The relative expressing level of SFRP5 protein in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 27.19±0.03, 47.54±0.05 and 74.93±0.05, respectively, showing a statistical difference among the three groups (F=183.49, P<0.001), and the relative expressing level of SFRP5 in the si-MeCP2 group was significantly higher than that in the MeCP2-mimic group and MeCP2-NC group (both at P<0.001). Conclusions MeCP2C can promote EMT of human LECs by down-regulating the expression of SFRP5 and therefore activating the Wnt3a/β-catenin signal pathway.

Objective To evaluate the effect of sevoflurane preconditioning on high-mobility group box 1 protein ( HMGB1) ∕Toll-like receptor 4 ( TLR4) ∕nuclear factor kappa B ( NF-κB) signaling pathway during lung ischemia-reperfusion ( I∕R) in rats. Methods Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-250 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group ( group S) , lung I∕R group ( group I∕R) and sevoflu-rane preconditioning group ( group SP ) . The right pulmonary hilum was only isolated but not ligated in group S. Lung I∕R was induced by clamping the right pulmonary hilum for 60 min followed by 120 min of reperfusion in anesthetized rats in group I∕R. In group SP, 2. 1％ sevoflurane was inhaled for 30 min to per-form sevoflurane preconditioning, and the lung I∕R model was established at 10 min after the end of inhala-tion. The rats were sacrificed at 120 min of reperfusion, and the lungs were removed for examination of the pathological changes which were scored and for determination of wet to dry weight ratio ( W∕D ratio) , con-tent of tumor necrosis factor-alpha ( TNF-α) in lung tissues ( by enzyme-linked immunosorbent assay) and expression of HMGB1, TLR4 and NF-κB protein in lung tissues (by Western blot). Results Compared with group S, the pathological scores, W∕D ratio and content of TNF-α were significantly increased, and the expression of HMGB1, TLR4 and NF-κB was up-regulated in I∕R and SP groups ( P<0. 05) . Compared with group I∕R, the pathological scores, W∕D ratio and content of TNF-αwere significantly decreased, and the expression of HMGB1, TLR4 and NF-κB was down-regulated ( P<0. 05) , and the pathological changes of lung tissues were significantly attenuated in group SP . Conclusion Sevoflurane preconditioning reduces lung I∕R injury probably through inhibiting HMGB1∕TLR4∕NF-κB signaling pathway in rats.

Objective To investigate the effects of perioperative parecoxib sodium on serum surfactant protein A and inflammatory response in elderly patients undergoing video-assisted thoracoscopic pneumonectomy,Methods Sixty-two ASA Ⅰ or Ⅱ elderly patients,aged 65-78 years,weighing 51-79 kg,scheduled for elective video-assisted thoracoscopic pneumonectomy under general anesthesia,were randomly divided into 3 groups:0.3 mg/kg parecoxib sodium group (group P1,n=21),0.6 mg/kg parecoxib sodium group (group P2,n =21) and control group (group C,n =20).The patients were given intravenous parecoxib sodium of 0.3 mg/kg immediately before induction of anesthesia and at 12 h after operation in group P1,and also parecoxib sodium of 0.6mg/kg immediately before induction of anesthesia and at 12 h after operation in group P2,while the equal volume of normal saline was given in group C.Blood samples were taken from the central vein before the induction of anesthesia(T0),after operation(T1),12 h after operation(T2) and 24 h after operation(T3).The concentration of serum surfactant protein A (SP-A),TNF-α,IL-6 and IL-8 were determined by ELASA.The incidence of pulmonary complications at 72 h after operation were also recorded.Results Compared with T0,the concentration of serum SP-A,TNF-α,IL-6 and IL-8 increased significantly in all groups at T1-T3 (P＜0.05).Compared with C group,the concentration of serum SP-A,TNF-α,IL-6 and IL-8 in groups P1 and P2 decreased significantly at T1-T3 (P＜0.05),there were no significant differences between groups P1 and P2.The incidence of postoperative pulmonary complications had no statistically significant differences between the three groups.Conclusion Parecoxib sodium can significantly reduce the concentration of serum SP-A and alleviate the inflammatory response in elderly patients undergoing video-assisted thoracoscopic pneumonectomy.

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway in propofol-induced reduction of lung ischemia-reperfusion (I/R) injury in aged rats.Methods Thirty-two clean healthy male Sprague-Dawley rats,aged 18-22 months,weighing 450-600,were divided into 4 groups (n =8 each) using a random number table:sham operation group (group S),I/R group (group I/R),I/R plus propofol group (group I/R+P) and all-trans retinoic acid (ARTA) plus I/R plus propofol group (group ARTA+I/R+P).Lung I/R was induced by occlusion of the right hilum of lung for 60 min followed by 120 min of reperfusion in anesthetized rats.In group ATRA+I/R+P,Nrf2/ARE signaling pathway blocker ARTA 6 mg/kg was intraperitoneally injected once a day for 3 consecutive days,and the model of lung I/R injury was established at 2 h after the last administration.In group I/R+P and group ARTA+I/R+P,while the model of lung I/R injury was established,propofol 30 mg · kg-1 · h-1 was infused via the caudal vein until 120 min of reperfusion.The rats were sacrificed at 120 min of reperfusion and then the lungs were removed for examination of the pathological changes which were scored and for measurement of wet/dry weight ratio (W/D ratio),superoxide dismutase (SOD) activity (by xanthine oxidase method),malondialdehyde (MDA) content (using thiobarbituric acid method) and expression of Nrf2 and heme oxygenase-1 (HO-1) protein (by Western blot).Results Compared with group S,the pathological scores,W/D ratio and MDA content were significantly increased,and the activity of SOD was decreased in I/R and ATRA+I/R+P groups,and the expression of Nrf-2 and HO-1 was significantly up-regulated in I/R,I/R+P and ATRA+I/R+P groups (P＜0.05).Compared with group I/R,the pathological scores,W/D ratio and MDA content were significantly decreased,the activity of SOD was increased,and the expression of Nrf-2 and HO-1 was up-regulated in group I/R+P (P＜0.05),and no significant change was found in the indexes mentioned above in group ARTA+I/R+P (P＞0.05).Compared with group I/R+P,the pathological scores,W/D ratio and MDA content were significantly increased,the activity of SOD was decreased,and the expression of Nrf-2 and HO-1 was down-regulated in group ARTA+ I/R+P (P＜0.05).Conclusion Nrf2/ARE signaling pathway activation is involved in propofol-induced reduction of lung I/R injury in aged rats.