Objective In this study,we analyzed the transcriptome sequences of Kupffer cells exposed to simulated microgravity for 3 d and conducted biological experiments to determine how microgravity initiates apoptosis in Kupffer cells. Methods Rotary cell culture system was used to construct a simulated microgravity model.GO and KEGG analyses were conducted using the DAVID database.GSEA was performed using the R language.The STRING database was used to conduct PPI analysis.qPCR was used to measure the IL1B,TNFA,CASP3,CASP9,and BCL2L11 mRNA expressions.Western Blotting was performed to detect the level of proteins CASP3 and CASP 9.Flow cytometry was used to detect apoptosis and mitochondrial membrane cells.Transmission electron microscopy was used to detect changes in the ultrastructure of Kupffer cells. Results Transcriptome Sequencing indicated that simulated microgravity affected apoptosis and the inflammatory state of Kupffer cells.Simulated microgravity improved the CASP3,CASP9,and BCL2L11 expressions in Kupffer cells.Annexin-V/PI and JC-1 assays showed that simulated microgravity promoted apoptosis in Kupffer cells.Simulated microgravity causes M1 polarization in Kupffer cells. Conclusion Our study found that simulated microgravity facilitated the apoptosis of Kupffer cells through the mitochondrial pathway and activated Kupffer cells into M1 polarization,which can secrete TNFA to promote apoptosis.

Objective:To review the characteristics and coping strategies of the rescue and landing medical support mission of Shenzhou-14 manned spacecraft.Methods:The characteristics of rescue and landing medical support mission of Shenzhou-14 manned spacecraft was analyzed, and the coping strategies and experience were discussed.Results:(1) The characteristics of rescue and landing medical support mission of Shenzhou-14 manned spacecraft included: long time in space station and high-intensity space missions; high-density space medical support mission in short term; special environmental factors in severe cold night; complex terrain of landing site; and the young medical support team. (2) The main coping strategies of rescue and landing medical support mission of Shenzhou-14 manned spacecraft included: strengthened the organization and leadership and improved the training model; reinforcement learning the medical treatment plan and strengthened the medical rescue skills training; optimized the carrying equipment and added the heat preservation and lighting measures; improved the medical rescue process and perfected the emergency plan; emphasized on the scientific research as important as mission; and strengthened the physical exercise and cold resistance exercise.Conclusions:The characteristics and coping strategies of rescue and landing medical support mission of Shenzhou-14 manned spacecraft are summarized to provide the experience for space medical rescue and offer the support for China's manned space industry.

Objective:To compare the clinical effecacy of artificial dermal regeneration matrix (as "artificial dermis" hereafter) and flap transfer in the treatment of soft tissue defects around interphalangeal joint.Methods:Through postoperative follow-up at outpatient clinic, a retrospective study was conducted on the clinical data of 60 patients who had soft tissue defects around the interphalangeal joints and received treatment in the Department of Hand Surgery, the Sixth Hospital of Ningbo from January 2018 to January 2022. According to applied surgical procedures, patients were divided into a flap transfer group (group A, n=30) and an artificial dermis group (group B, n=30). Group A included 19 males and 11 females aged 44.83 years ± 11.56 years including 5 patients with simple soft tissue defects, and 6 with soft tissue defects and fractures, 10 with soft tissue defects with tendon and (or) ligament injuries, 3 with soft tissue defects and vessel and (or) nerve injuries, and 6 with soft tissue defects and over 2 other types of compound injuries. The defect areas ranged from 2.5 cm×1.2 cm to 5.0 cm×1.6 cm. After emergency debridement and treatment for bone and tendon injuries, the patients in group A received transfers of free flap or transfers with adjacent digit flaps or island flaps. Postoperative functional exercise started from 1-6 weeks after surgery. Patients in group B included 17 males and 13 females, aged 44.70 years ± 11.20 years and there were 6 patients with simple soft tissue defects, 6 with soft tissue defects and fractures, 9 with soft tissue defects and tendon and (or) ligament injuries, 5 with soft tissue defects and vessel and (or) nerve injuries, and 4 with soft tissue defects and over 2 other types of compound injuries. The defect area ranged from 3.1 cm ×1.3 cm to 4.5 cm × 1.8 cm. Debridement and treatment of the bone and tendon injury in group B were the same as what in group A, but the patients in group B received artificial dermis coverage other than transfer of flap. After an artificial dermis had completely vascularised, a split-thickness skin graft was performed over the neo-dermis in the second phase surgery. Postoperative functional exercise started from 1-2 weeks after artificial dermis grafting surgery. The interval time of flap transfer or split-thickness skin grafting, survival rate of flap transfer or split-thickness skin grafting, Vancouver Scar Scale(VSS), TPD and total active movement(TAM) were compared between the 2 groups. The count data were analysed by Chi-square test. All measured data were analysed by independent sample t test or Mann-Whitney U test. P<0.05 indicates a statistically significant difference. Results:The interval time of flap transfer in group A and artificial dermis grafting in group B were 2.93 days ± 2.48 days and 19.87 days ± 3.35 days, respectively. There was a statistically significant difference between the 2 groups( P<0.05). All patients were entered in postoperative outpatient follow-up for 11-14 months(12.00 months ± 0.93 months). The appearance of flaps of some patients in group A was bloated with slightly limited digit movement. In group B, the digit movement was normal, without obvious scar hyperplasia in both of the recipient and donor sites. The survival rate of flap transfer or artificial dermis graft, TPD, VSS score and TAM score for group A and group B were 96.00% ± 9.32% and 98.17% ± 3.07%, respectively, 8.67 mm ± 2.01 mm and 9.50 mm ± 1.81 mm, 3.40 ± 1.07 and 3.17 ± 0.91 and 18.30 ± 1.97 and 18.93 ± 1.64, respectively. There were no significant differences between the 2 groups( P>0.05). Conclusion:In comparison with the transfer of flap, an artificial dermis is also effective and satisfactory in the treatment of soft tissue defects around the interphalangeal joint. Further large scale and multi-centre investigations are required.

OBJECTIVES: To provide reference basis for reducing the mortality for children under 5 years old and promote the healthy development, the mortality for children under 5 years old and the main causes for death in Liuyang City from 2013 to 2020 are analyzed. METHODS: The data of 725 cases of death for children under 5 years old in Liuyang City from 2013 to 2020 were collected.The causes and difference of death among the children were analyzed retrospectively by descriptive statistic methods. RESULTS: There were a total of 144 516 live births in Liuyang City from 2013 to 2020. The mortality for children under 5 years old was 5.01‰, for infants was 3.39‰, and for newborns was 1.63‰. The male child mortality was 5.28‰, and the female child mortality rate was 4.72‰, with significant difference (P>0.05). The mortality for children under 5 years old was seasonal fluctuation, without significant difference among seasons (P>0.05). For the past 5 years, the top 3 causes for death among children under 5 years old were preterm birth and low birth weight, congenital heart disease, and pneumonia. Before death, 341 cases (47.04%) were treated in provincial hospitals, 198 cases (27.31%) in county-level hospitals, 56 cases (7.72%) in village-level hospitals, and 130 cases (17.93%) were not treated. CONCLUSIONS The mortality for children under 5 years old in Liuyang City is gradually reduced in the past 5 years. The main causes for death are premature birth and low birth weight, congenital heart disease and pneumonia. We should develop healthy education, improve the rate of prenatal diagnosis, promote the construction of obstetrics and paediatrics, and fundamentally reduce the mortality for children under 5 years old.
Cause of Death ; Child ; Child Mortality ; Child, Preschool ; Female ; Heart Defects, Congenital ; Humans ; Infant ; Infant Mortality ; Infant, Newborn ; Male ; Pneumonia/epidemiology* ; Pregnancy ; Premature Birth ; Retrospective Studies

Objective:To study the environmental characteristics of launch section during the space medical rescue mission of Shenzhou-13 spacecraft, evaluate the influence of special environment on peripheral veins, and investigate the coping strategies.Methods:Combined with the practice of launch section during space medical rescue mission of Shenzhou-13 spacecraft, the main factors of aerospace special environment, cold and faint light, were selected and the related literature was reviewed to explore the feasibility, advantages and disadvantages of peripheral venipuncture, central venous catheter and bone marrow puncture. The experience of the space medical rescue mission was summarized and the coping strategies were provided for follow-up task training.Results:The environmental characteristics of launch section during the space medical rescue mission of Shenzhou-13 spacecraft included cold, faint light, sand-wind and complex terrain. The vascular filling, vascular elasticity and visibility of peripheral vein decreased in different degrees under special environmental, the vascular visibility was most affected. The literature analysis showed that bone marrow puncture was a suitable method to establish infusion access in cold and faint light environment.Conclusions:The special environment of space launch mission has greater impact on peripheral veins. In order to assurance the successful implementation of manned space project, the training of basic operational skills should be consolidated, the training of ability in special environment should be enhanced and the advanced clinical auxiliary equipment should be equipped in follow-up space medical mission training.

Pancreatoduodenectomy (PD) is a very challenging operation in pancreatic surgery. Robotic surgery system has obvious advantages in precise operation. In specific patients, minimally invasive (laparoscopic and robotic surgery system) PD is less invasive than conventional open PD, showing good safety and feasibility. There are some differences between robotic surgery system and laparoscopic system in surgical field of vision, surgical operation and anastomosis mode. The authors review the relevant studies and conduct a Meta-analysis to evaluate the perioperative efficacy of robotic versus open approach for PD.

ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.

Objective:To observe the effect of Lingshao-Zaoren Decoction on urodynamics and the expression of Piezo1 if overactive bladder (OAB) rats. Methods:Thirty SPF grade female SD rats were randomly divided into blank group, model group, Tolterodine control group, low-dose and high-dose Lingshao-Zaoren Decoction groups, with 6 rats in each group. The OAB rats were modeled by intraperitoneal injection of Cyclophosphamide. After the successful modeling, Tolterodine control group was given 0.36 mg/kg Tolterodine tartrate, the low-dose and high-dose Lingshao-Zaoren Decoction groups were given 1.59 and 3.18 g/kg Lingshao-Zaoren Mianjian granules by gavage, the blank group and model group were given the same amount of distilled water, once a day for 14 days. After 14 days, the urodynamics of rats in each group were detected. The bladder volume and maximum bladder pressure were observed respectively. The pathological changes of bladder tissue were observed by HE staining. The expression of Piezo1 protein in bladder tissue was detected by immunohistochemistry and Western blot. The expression of Piezo1 mRNA in bladder tissue was detected by qPCR. Results:Compared with the blank group, the body weight, bladder volume and maximum bladder pressure of the model group were significantly reduced ( P<0.01). HE staining result showed that the model group had hyperplasia of urinary tract epithelium, degeneration, necrosis and abscission of epithelial cells, infiltration of a large number of inflammatory cells in stroma, vascular proliferation, thickening of vascular wall, hyperplasia of mucosal smooth muscle, disorder of arrangement, and significant up regulation of Piezo1 protein expression ( P<0.01). Compared with the model group, the weight [(244.83 ± 6.05) g, (233.33 ± 11.76) g vs. (219.00 ± 9.70) g] of rats in the Tolterodine control group and high-dose group of Lingshao-Zaoren Decoction significantly increased ( P<0.01), and the bladder volume [(0.93 ± 0.31) ml, (1.17 ± 0.17) ml, (1.21 ± 0.23) ml vs. (0.50 ± 0.16) ml] and maximum bladder pressure [(42.00 ± 3.03) cmH 2O, (45.83 ± 7.19) cmH 2O, (46.83 ± 8.23) cmH 2O vs. (30.50 ± 5.47) cmH 2O] of rats in the Tolterodine control group, low-dose and high-dose Lingshao-Zaoren Decoction groups were significantly increased ( P<0.01); the bladder epithelial hyperplasia and degeneration degree, interstitial inflammatory cell infiltration degree and vascular hyperplasia degree of rats in the Tolterodine control group, low-dose and high-dose Lingshao-Zaoren Decoction groups significantly increased. The expression of Piezo1 mRNA (1.50 ± 0.04, 2.05 ± 0.08, 1.44 ± 0.10 vs. 2.56 ± 0.11) and protein in the Tolterodine control group, low-dose and high-dose Lingshao-Zaoren Decoction groups were significantly decreased ( P<0.01). Conclusion:Lingshao-Zaoren Decoction can increase the bladder volume and maximum bladder pressure of urinary incontinence caused by detrusor overactivity in rats with overactive bladder, which may be related to reduction of Piezo1 expression.

Objective:To investigate the clinical effect of free superficial circumflex iliac artery flap combined with anterolateral thigh flap in repairing large skin defect of the hand.Methods:A retrospective case series study was conducted to analyze the clinical data of 9 patients with large skin defect of the hand admitted to Ningbo No.6 Hospital from January 2016 to December 2019，including 7 males and 2 females，aged 31-63 years［（45.3±5.6）years］. The area of skin defect reanged from 20 cm×15 cm to 25 cm×20 cm. One side of the superficial circumflex iliac artery flap and the contralateral anterolateral thigh flap were used for repair. Both recipient and donor sites were sutured at the first stage. The ilioinguinal region and thigh donor region were sutured directly. The area of superficial circumflex iliac artery flap was from 19 cm×6 cm to 23 cm ×10 cm and that of anterolateral thigh flap was 19 cm×9 cm to 23 cm×8 cm. The flap survival，healing of donor and recipient area and complications were observed. At 1，6 and 12 months after operation，the function of the flap was evaluated by the late functional evaluation criteria of the flap，hand function by the upper limb functional evaluation criteria of the Chinese Medical Association，and scar condition of the donor site by Vancouver Scar Scale（VVS）. At the last follow-up，the color，elasticity，appearance and scar of the donor site were observed.Results:All patients were followed up for 6-18 months［（12.5±2.3）months］. All flaps survived successfully. All patients achieved donor-site healing at the first stage，except that one patient experienced incision dehiscence in the thigh donor site after operation and healed after the second stage suture. The texture and appearance of the flaps were good. The flap in 5 patients was obviously bloated. Therefore，the secondary skin flap repair was selected，and the appearance was significantly improved after operation. Scores for function of the flap and the hand were improved over time（ P<0.05）. At 12 months after operation，the function of the flap was excellent in 3 patients，good in 4 and fair in 2，with the excellent and good rate of 78%；the result of hand function was excellent in 3 patients，good in 3 and fair in 3，with the excellent and good rate of 67%. The VVS score of the donor site decreased from（9.7±1.3）points at postoperative 1 month to（5.7±0.9）points at postoperative 6 months and（3.4±0.8）points at postoperative 12 months（ P<0.05）. At the latest follow-up，the color and elasticity of the flap was similar to that of the surrounding skin tissue，only with slight bloating；the scar of the donor site was small. Conclusions:The free superficial circumflex iliac artery flap combined with anterolateral thigh flap can cover large skin defect of the hand at one time，with good appearance and texture of the flap. The appearance and function of the wounded hand are good after operation，leaving only linear scar in the donor site.

Objective:To investigate the clinical application and effect of end-to-side anastomosis in personalised free ilioinguinal flap transfer.Methods:From March, 2015 to July, 2020, 88 patients with soft tissue (bone) defect of limbs were treated. Different ilioinguinal flaps were designed according to the wound condition of patients, which were 48 cases of free superficial circumflex iliacartery perforator flap, 7 cases of free superficial epigastric artery perforator flap, 19 cases of composite tissue flap with iliac bone, 8 cases of combined flap of superficial circumflex iliac artery and superficial abdominal wall artery, and 6 cases of superficial circumflex iliac artery and superficial abdominal wall artery lobulated flap. The area of the flap was 4.0 cm×6.0 cm-10.0 cm×30.0 cm. The artery and recipient artery were anastomosed end-to-side: 36 cases to radial artery; 12 cases to ulnar artery; 18 cases to dorsalis pedis artery; 15 cases to anterior tibial artery; 7 cases to posterior tibial artery. Venous anastomosis of skin flap: 42 cases were anastomosed with 2 veins, which were superficial vein of the same name and accompanying vein; 46 cases were anastomosed with 1 superficial vein of the same name. The accompanying vein of the flap was anastomosed end-to-side with the accompanying vein of the main artery of the recipient area, and the superficial vein of the same name was anastomosed end-to-end with the accompanying vein or subcutaneous superficial vein of the recipient artery. Follow-up includes flap blood supply, blood supply to the distal limbs, appearance of both the donor site and the recipient area, and patient satisfaction.Results:There were 83 cases of flaps survived successfully, and 5 cases of crisis. Among them, 2 cases had artery crisis at 48 h after surgery. After exploration, it was found that 1 case caused by arterial thrombosis, and 1 case compressed by the stapler that anastomoses the vein. The other 3 cases had venous crisis at 72 h after surgery: after exploration, it was found that caused by thrombosis at the venous anastomotic site. The average follow-up period was 10 (range, 3-24) months. All flaps survived after re anastomosis or vascular transposition. The donor site and recipient site of the flap healed well. The blood supply of the flap was good and the texture was soft. There was no blood supply disorder in the distal limb.Conclusion:The end-to-side anastomosis technique is suitable for all kinds of free flap transplantation in ilioinguinal region, with high vascular patency rate. It can not only solve the problem of thin vascular pedicle of donor site flap, but also retain the main artery of recipient limb without affecting the distal blood supply.