Objective To investigate the effect and mechanism of miR-20a-5p on human nephroblastoma cell line WiT49 transplanted tumor in nude mice.Methods The gene expression chip was downloaded from GEO database,and the differential gene miR-20a-5p was obtained by GEO2R.The NF-κB gene was positively correlated with the expression of miR-20a-5p through cBioPortal database.The target gene of miR-20a-5p was predicted to be NFKBIB of the NF-κB transcription factor suppressor protein family by targetscan database,and was verified by dual luciferase assay.Nephroblastoma cell line WiT49 was cultured in vitro and transfected into WiT49 cells with lentiviral vectors constructed with miR-20a-5p mimics and its suppressor gene.Twelve nude mice were randomly divided into three groups:WiT49 model group,WIT49-miR-20a-5p overexpression group and WIT49-miR-20a-5p knockdown group.The tumor mass and volume of each group were detected by tumor formation experiment in nude mice.real time fluorescent quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-20a-5p,NFKBIB and NF-κB in each group;CCK-8 cell proliferation assay was used to verify the proliferation of tumor cells in each group.Results miR-20a-5p is highly expressed in nephroblastoma and is positively correlated with the expression of NF-κB.miR-20a-5p and NFKBIB have mutual binding sites and binding effects.In the tumor formation experiment of nude mice,the tumor volume and mass of WIT49-miR-20a-5P overexpression group were significantly increased compared with WiT49 model group,and the difference was statistically significant(P<0.05).In the qRT-PCR test,the expressions of miR-20a-5p and NF-κB in the WIT49-miR-20a-5p overexpression group were higher than those in the WiT49 model group,and NFKBIB expression in the WIT49-miR-20a-5p overexpression group was lower than that in the WiT49 model group,with statistical significance(P<0.05).CCK-8 cell proliferation assay showed that the absorbance of WIT49-miR-20a-5p overexpression group at 24 and 48 hours was higher than that of WiT49 model group,and the absorbance of WIT49-miR-20a-5p knockdown group at 24,48 and 72 hours was lower than that of WiT49 model group,and the difference was statistically significant(P<0.05).Conclusion miR-20a-5p may promote the growth of human nephroblastoma cell WiT49 transplanted tumor in nude mice by regulating NFKBIB activation of NF-κB pathway.

Objective:To investigate the influences and mechanism of extracellular vesicles from dermal papilla cells (DPC-EVs) of mice on human hypertrophic scar fibroblasts (HSFs).Methods:The study was an experimental research. The primary dermal papilla cells (DPCs) of whiskers were extracted from 10 6-week-old male C57BL/6J mice and identified successfully. The DPC-EVs were extracted from the 3 rd to 5 th passage DPCs by ultracentrifugation, and the morphology was observed through transmission electron microscope and the particle diameter was detected by nanoparticle tracking analyzer ( n=3) at 24 h after culture. The 3 rd passage of HSFs were divided into DPC-EV group and phosphate buffer solution (PBS) group, which were cultured with DPC-EVs and PBS, respectively. The cell scratch test was performed and cell migration rate at 24 h after scratching was calculated ( n=5). The cell proliferation levels at 0 (after 12 h of starvation treatment and before adding DPC-EVs or PBS), 24, 48, 72, and 96 h after culture were detected by using cell counting kit 8 ( n=4). The protein expressions of α-smooth muscle actin (α-SMA) and collagen typeⅠ (ColⅠ) in cells at 24 h after culture were detected by immunofluorescence method and Western blotting, and the protein expression of Krüppel-like factor 4 (KLF4) in cells at 24 h after culture was detected by Western blotting. After the 3 rd passage of HSFs were cultured with DPC-EVs for 24 h, the cells were divided into blank control group, KLF4 knockdown group, and KLF4 overexpression group according to the random number table. The cells in blank control group were only routinely cultured for 48 h. The cells in KLF4 knockdown group and KLF4 overexpression group were incubated with KLF4 knockdown virus for 24 h, then the cells in KLF4 knockdown group were routinely cultured for 24 h while the cells in KLF4 overexpression group were incubated with KLF4 overexpression virus for 24 h. The protein expressions of KLF4, α-SMA, and ColⅠ in cells were detected by Western blotting at 48 h after culture. Results:At 24 h after culture, the extracted DPC-EVs showed vesicular structure with an average particle diameter of 108.8 nm. At 24 h after scratching, the migration rate of HSFs in PBS group was (54±10)%, which was significantly higher than (29±8)% in DPC-EV group ( t=4.37, P<0.05). At 48, 72, and 96 h after culture, the proliferation levels of HSFs in DPC-EV group were significantly lower than those in PBS group (with t values of 4.06, 5.76, and 6.41, respectively, P<0.05). At 24 h after culture, the protein expressions of α-SMA and ColⅠ of HSFs in DPC-EV group were significantly lower than those in PBS group, while the protein expression of KLF4 was significantly higher than that in PBS group. At 48 h after culture, compared with those in blank control group, the protein expression of KLF4 of HSFs in KLF4 knockdown group was down-regulated, while the protein expressions of α-SMA and ColⅠ were both up-regulated; compared with those in KLF4 knockdown group, the protein expression of KLF4 of HSFs in KLF4 overexpression group was up-regulated, while the protein expressions of ColⅠ and α-SMA were down-regulated. Conclusions:The DPC-EVs of mice can inhibit the proliferation and migration of human HSFs and significantly inhibit the expressions of fibrosis markers α-SMA and ColⅠ in human HSFs by activating KLF4.

Objective To investigate the risk factors of Hirschsprung's associated enterocolitis (HAEC) to provide a basis for the diagnosis and treatment of Hirschsprung's disease (HSCR),and offer the guidance for reducing the postoperative complications occurrence and postoperative rehabilitation. Methods The case data of the children patients with HSCR treated by initial surgery in the pediatric surgery department of this hospital from January 2015 to January 2023 were analyzed retrospectively.The risk factors of HAEC occurrence were analyzed.Results Among 110 children patients with HSCR,34 cases developed HAEC with a incidence rate of 30.91％.There were statistical differences in different ages,preoperative enter-itis occurrence,low body weight,length of spastic segment of lesion and diet control (P<0.05).The multiva-riate logistic regression analysis results showed that the age ≥6 years old (OR=2.407,95％CI:1.040-5.568),preoperative bowel washing time<2 weeks (OR=2.464,95％CI:1.082-5.612),preoperative enter-itis occurrence (OR=2.943,95％CI:1.193-7.260),low body weight (OR=2.588,95％CI:1.113-6.014),and the length of spastic segment of lesion ≥30 cm (OR=2.801,95％CI:1.224-6.412) were the risk fac-tors of HAEC occurrence (P<0.05),while dietary control (OR=0.358,95％CI:0.153-0.837) was the pro-tective factor for HAEC (P<0.05).Conclusion Prevention after operation in the children patients with HSCR should be carried out according to the risk factors of HAEC occurrence.

Objective:To evaluate the safety and efficacy of enhanced recovery after surgery(ERAS) in the perioperative period of congenital choledochal cysts in children.Methods:This is a multicenter prospective randomized controlled study. The clinical data of 273 pediatric congenital choledochal cysts(CCC) patients who underwent surgery at 14 medical centers with complete follow-up data were collected through the medical data analysis platform. Among them, 123 cases in ERAS group were managed perioperatively in strict accordance with ERAS mode, and 150 cases in conventional group were managed according to traditional mode. The length of hospital stay,time to first farting, time to complete feeding, the incidence of complications, cost and readmission rate within 30 days,stress indexes and liver function were compared between the two groups.Results:Compared with the conventional group, median time to start farting (2.0 d vs. 3.0 d, P<0.001), median time to complete feeding (5.0 d vs. 7.0 d, P<0.001), median postoperative hospitalization time (6.0 d vs. 9.0 d, P<0.001),the median total length of stay(13.0 d vs. 15.0 d, P<0.001) were shorter,the median hospitalization cost (37,000 yuan vs.43,000 yuan P<0.001) was lower, and stress indexes recovered quickly. The incidence of postoperative hospital stay and readimission rate within 30 d were not statistically different between the two groups. Conclusion:It is safe and feasible to implement ERAS for children with CCC in the perioperative period, which can reduce stress response, speed up recovery,and save medical costs.

Type 2 diabetes mellitus (T2DM) is a common chronic disease characterized by hyperglycemia. Itchy skin is common in patients with diabetes, and high blood sugar leads to decreased immunity and nerve damage, causing skin problems. 40 patients with T2DM complicated with pruritus diagnosed and treated in our hospital from March 2022 to September 2022 were included to observe the clinical characteristics of the patients.

【Objective】 To explore the efficacy of Mathieu combined with tongue-shaped flap covering in children with distal hypospadias and micropenis head deformity. 【Methods】 The clinical data of 72 patients with distal hypospadias complicated with micropenis treated during Jan.2018 and Jun.2021 were retrospectively analyzed, including 35 patients who underwent modified Mathieu combined with tongue-shaped flap and external urethral orificium (study group), and 37 patients who underwent traditional Mathieu (control group). Urethral stricture, urethral fistula, urethral diverticulum and penile head cleft were compared between the two groups. 【Results】 Urinary fistula occurred in 2 cases (5.27%) and 9 cases (24.32%) in the study group and control group, respectively, the incidence being much lower in the study group (P=0.028). Urethral stricture occurred in 1 case (2.86%) and 1 case (2.70%) in the study group and control group, respectively; penile head dehiscence in 1 case (2.86%) and 3 cases (8.11%); urethral diverticulum in 0 case (0%) and 2 cases (5.41%). There were no significant differences between the two groups in the incidence of urethral stricture, urethral dehiscence and urethral diverticulum (P>0.05). 【Conclusion】 Mathieu combined with tongue-shaped flap covering to treat children with distal hypospadias with micropenis head deformity can reduce the incidence of postoperative complications, achieve high surgical satisfaction and appearance satisfaction in the first phase, reduce harm caused by surgery, and promote patients’ psychological health.

OBJECTIVE：To preliminarily investigate the improveme nt effects and mechanism of Mongolian medicine Saorilao-4 decoction on specific pulmonary fibrosis model rats. METHODS ：Male SD rats were randomly divided into normal control group ，model group ，positive control group （pirfenidone，0.163 g/kg）and Saorilao- 4 decoction low ，medium and high dose groups （0.899，1.798，3.596 g/kg），8 rats in each group. Except for normal control group ，other groups were given 6 mg/mL bleomycin intratracheally at 5 mg/kg once to induce the specific pulmonary fibrosis model. From the first day after modeling ， normal control group and model group were given normal saline intragastrically ，other groups were given corresponding drugs intragastrically，once a day ，10 mL/kg，for 4 weeks. During the experimental period ，the general condition of the rats in each group was observed and the body mass was weighed. Twenty-four h after last medication ，the appearance morphology of rat l ung in each group were observed. The morphological characteristics of lung tissues were observed by HE and Masson staining. ELISA was adopted to determine the activity of SOD and the content of MDA in serum ，the contents of hydroxyproline （HYP），IL-1β，IL-6，hyaluronidase（HA），laminin（LN）precollagen type Ⅲ（PC-Ⅲ）and collagen type Ⅳ（Col-Ⅳ）in lung tissue. RT-PCR was used to determine the mRNA 发。E-mail：bwf007007@sina.com expression of TGF-β 1，Smad3 and Smad 7 in lung tissue. RESULTS：Compared with model group ，the activity ，hair and diet of the rats in each dose group of Saorilao- 4 decoction and positive control group were significant ly improved ，and the body mass after the last administration was significantly increased ； the pathological change of lung and pulmonary fibrosis were significantly improved ，and the activity of SOD in serum was increased significantly. Serum content of MDA （except for Saorilao- 4 decoction medium dose group ），the contents of HYP （except for Saorilao- 4 decoction high dose group ），IL-1β，IL-6，HA，LN，PC-Ⅲ，Col-Ⅳ（except for Saorilao- 4 decoction high dose group）as well as mRNA expression of TGF-β1 and Smad 3 in lung tissue were significantly decreased ；mRNA expression of Smad 7 was significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Saorilao-4 decoction can significantly improve the lung pathological changes ，delay and reverse the progression of pulmonary fibrosis in specific pulmonary fibrosis model rats ，the mechanism of which may be related to the inhibition of inflammatory response ， improvement of lipid peroxidation ， down-regulation of TGF-β1 and Smad 3 mRNA expression ，and up-regulation of Smad 7 mRNA expression.

OBJECTIVE：To investigate inhibitory effects of Qing fei baoyu an capsule on airway inflammation in rats with chronic obstructive pulmonary disease （COPD），and its effects on NLRP 3 signaling pathway. METHODS ：Totally 60 SD male rats were randomly divided into blank control group ，model group ，dexamethasone group （positive control ，0.2 mg/kg），Qingfei baoyuan capsule high-dose ，medium-dose and low-dose groups （1 232.0，616.0，308.0 mg/kg），with 10 rats in each group. Except for blank control group ，other groups were fumigated for 28 days and given intratracheal dripping of lipopolysaccharide twice to induce COPD model. Since the 29th day after modeling ，blank control group and model group were given constant volume of normal saline intragastrically ，and administration groups were given related medicine intragastrically. The administration volume was 10 mL/kg，once a day ，for consecutive 28 days. After last administration ，the lung function was detected. The pathological changes of lung tissue were observed by HE staining. The content of interleukin- 1β（IL-1β）in bronchoalveolar lavage fluid （BALF）were detected by ELISA ，and the number of leukocytes was counted ；the expression of NLRP 3 and Cleaved caspase- 1 in lung tissue of rats were detected by Western blotting assay. RESULTS：Three，two，one，one and two rats died in model group， dexamethasone group ， Qingfei baoyuan capsule high-dose，medium-dose and low-dose groups ，respectively. Compared with blank control group ，FEV0.3/FVC of rats in # model group was significantly decreased （P＜0.01）. A large number of inflammatory cells infiltration we re found in the lung tissue ，and lung tissue lesion was obvious. The content of IL- 1β and white blood cell count in BALF，relative expression of NLRP3 and Cleaved caspase- 1 protein in lung tissue were increased significantly （P＜0.05 or P＜0.01）. Compared with model group，FEV0.3/FVC of administration groups were increased significantly （P＜0.05 or P＜0.01）；lung tissue lesion of them were improved to different extents. The content of IL- 1β and white cell count in BALF，relative expression of NLRP 3 protein（except for Qingfei baoyuan capsule low-dose group ）and Cleaved caspase- 1 protein（except for Qingfei baoyuan capsule medium-dose and low-dose groups ）in lung tissue were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Qingfei baoyuan capsule can relieve lung tissue lesion and improve lung function in COPD model rats ，the effects of which may be associated with inhibiting inflammation reaction by inhibiting NLRP 3 signaling pathway.

Objective:To investigate the clinical application effect of latissimus dorsi muscle flap in reconstruction of muscle strength around shoulder after electric burns.Methods:From March 2014 to September 2020, 13 patients with electric burns and severe injury around shoulder were admitted to the First Affiliated Hospital of Air Force Medical University, including 11 males and 2 females, aged 19-55 years. A retrospective observational study was conducted. The left upper limbs were injured in 8 cases, and the right upper limbs were injured in 5 cases, all with eschar wounds of Ⅲ-Ⅳ degree. Among which, there were biceps defects in 6 cases, deltoid defects in 3 cases, triceps defects in 2 cases, and composite defects of multiple muscles around shoulder in 2 cases. The surgery was carried out in two stages. In stage Ⅰ, debridement and exploration of electric burn wounds around shoulders were conducted to preserve local tissue and save the limb as much as possible on the premise of guaranteeing the stability of the body condition. After the last debridement, the wound area was from 10 cm×6 cm to 40 cm×15 cm, the muscle defect area was from 8 cm×4 cm to 19 cm×12 cm, and the humerus was exposed in 7 patients. In stage Ⅱ, according to the residual limb defect degree, muscle reconstruction around shoulder was conducted with the latissimus dorsi muscle flap, and area of the latissimus dorsi muscle flap was 15 cm×6 cm to 20 cm×18 cm. The residual wounds were repaired with autologous split-thickness skin grafts of head, and the donor sites of muscle flaps were sutured directly. The survivals of the muscle flaps and wounds closure post operation, and the appearances of the donor sites and recipient sites during follow-up were observed. At the last follow-up, the shoulder joint function was evaluated using the trial standard for the evaluation of the functions of the upper limbs of the Hand Surgery Society of the Chinese Medical Association, and the satisfaction degrees of patients for appearance and function recoveries of shoulder were investigated by self-made questionnaire with reference to the concise test scoring system of shoulder joint.Results:All of the 13 muscle flaps around shoulder survived after surgery. Two patients had residual wounds in the skin grafting area, the wound in one of the patients was healed after dressing change, and the wound in the other 1 patient was healed with the second autologous split-thickness skin grafting on head after dressing change. During follow-up of 6 to 18 months for all the patients, the muscle flaps of patients were full in appearance and not bloated, and atrophic scar in the repaired area was soft in texture and closed with normal skin around. Linear suture scars were left in the donor sites of muscle flaps, which did not affect the overall appearance. At the last follow-up, the active abduction range of the shoulder joint was 60-90°, upward lift on 120-180°, muscle strength recovered to level Ⅳ and above in 8 cases and to level Ⅲ in 5 cases, and the shoulder joint function was evaluated as excellent in 8 cases and good in 5 cases; 10 patients were very satisfied and 3 patients were satisfied with the appearance and function recovery of the shoulders.Conclusions:The application of latissimus dorsi muscle flap provides a better choice for the muscle strength reconstruction around shoulder after electric burns, with good appearance of the operative areas and ideal prognosis of upper limb function.

Objective:To compare the effect of posterolateral plate and anterolateral locking plate in surgical treatment for extra-articular distal humeral shaft fractures.Methods:A retrospective case-control study was made on 52 patients with extra-articular distal humeral shaft fractures admitted in Tianjin Hospital from January 2014 to October 2016. There were 32 males and 20 females, with the age from 18 to 56 years [(36.9±10.9)years]. According to the AO/OTA classification, there were 15 patients with type A, 32 type B, and 5 type C. Twenty-three patients were treated with osterolateral locking plate (Group A) and 29 with anterolateral locking plate (Group B). Operation time, bone union time, range of motion of the elbow, Mayo elbow performance score and complication rate were evaluated.Results:All patients were followed up for 12-20 months [(13.7±2.2)months]. Operation time was (79.8±9.6)minutes in Group A and (85.0±11.6)minutes in Group B ( P>0.05). Bone union time was (4.1±1.0)months in Group A and (4.1±1.0)months in Group B ( P>0.05). Degrees of elbow extension was 3.9°(0.0°, 5.0°) in Group A, and 4.4°(0.0°, 5.0°) in Group B ( P>0.05). Degrees of elbow flexion was 127.4°(125.0°, 132.50°) in Group A and 128.5°(122.5°, 132.5°) in Group B ( P>0.05). Mayo elbow performance score was 91.0 (90.0, 93.5) points in Group A and 90.2 (90.0, 92.5)points in Group B ( P>0.05). Radial nerve damage was noted after operation, showing no significant difference between two groups [2 patients (9%) in Group A and 3 patients (10%) in Group B] ( P>0.05), and the symptoms were recovered in all patients within 3 months. Conclusions:Both the posterolateral and anterolateral locking plate are effective in surgical treatment for extra-articular distal humeral shaft fractures. However, the posterolateral plate can be placed closer to the distal end of the humerus and the multi-directional locking design is more stable, which has advantages for the patient with fracture line close to the elbow joint.