AIM:To investigate the possible mechanism how orphan nuclear receptor 4A1(NR4A1)affects renal fibrosis in unilateral ureteral obstruction(UUO)mice.METHODS:(1)Specific pathogen-free(SPF)male C57BL/6J mice(5 to 6 weeks old)were randomly divided into sh-Con+sham group(n=6),sham+sh-NR4A1 group(n=6),sh-NR4A1+UUO group(n=6),and sh-NR4A1+UUO group(n=6).Renal NR4A1 knockdown mice were prepared by intrarenal injection of NR4A1 viral vector.(2)SPF male C57BL/6J mice(5 to 6 weeks old)were randomly assigned to in sham group,UUO group,and UUO+cytosporone-B(Csn-B)group.An animal model of renal fibrosis was prepared by UUO,and Csn-B was intervened for 14 days.HE,Masson,and Sirius red staining were used to observe renal pathological damage.Immunohistochemistry and Western blot were performed to detect the expression of NR4A1,interferon-γ(IFN-γ),α-smooth muscle actin(α-SMA)and vimentin.The purpose of this study is to observe the regulatory effect of NR4A1 on UUO-induced renal fibrosis.RESULTS:(1)The expression of NR4A1 was decreased in kidney tissues of UUO mice(P<0.05).(2)HE staining results showed that there are tubular dilation,atrophy,and massive inflammatory cell infiltra-tion in sh-Con+UUO group,and NR4A1 knockdown can aggravate UUO-induced kidney damage(P<0.05).The results of Masson and Sirius red staining showed a banded distribution of collagen deposition in the sh-Con+UUO group,and colla-gen deposition increased significantly after NR4A1 knockdown(P<0.05).Treatment with Csn-B could improve renal path-ological damage and reduce collagen deposition.(3)UUO could upregulate the expression of α-SMA and vimentin,and NR4A1 knockdown could significantly increase UUO-induced their expression(P<0.05).In addition,Csn-B could im-prove UUO-induced renal fibrosis(P<0.05).(4)The expression of IFN-γ was increased in UUO mice,and NR4A1 knockdown could upregulate UUO-induced IFN-γ expression(P<0.05).Moreover,Csn-B could down-regulate UUO-in-duced IFN-γ expression(P<0.05).CONCLUSION:NR4A1 can affect renal fibrosis by regulating IFN-γ in UUO mice.

Objective:Exploring the role and mechanism of CHMP4C in regulating necroptosis during gastric can-cer development and progression.Method:The expression of CHMP4C in pan-cancer was analyzed by bioinformatics methods,and the expression of CHMP4C was detected in human normal gastric epithelial cells and GC cell lines by RT-qPCR and Western blot.Overexpression or knockdown of CHMP4C was performed in GC cell lines,and the effects of CHMP4C on the growth and proliferation of GC cells were detected using CCK-8 and clone formation assays.The CCK-8 experiment and Hoechst/PI double staining experiment were used to detect the changes in GC cell mortality and PI positive cell ratio after treatment with the necroptsis inducer TSZ or inhibitor necrostatin-1(Nec-1).Western blot assay was used to detect the protein and phosphorylation levels of RIPK1,RIPK3,and MLKL in GC cells.Result:CHMP4C was upregulated in GC tissues and cells.The CCK-8 and clone formation experiments showed that overex-pression of CHMP4C significantly improved the proliferation ability and colony formation efficiency of GC cells,while knockdown of CHMP4C significantly weakened GC cells.Moreover,the results of CCK-8 and Hoechst 33342/PI double staining experiments showed that upregulated CHMP4C could inhibit TSZ induced GC cell death;Nec-1 can reverse the decrease in GC cell viability caused by CHMP4C knockdown.Western blot experiment showed that the levels of p-RIPK1,p-RIPK3,and p-MLKL were significantly decreased in overexpressing cells,while they were increased in knockdown cells.After treatment with Nec-1,the expression levels of these three proteins decreased in knockdown cells.Conclusion:CHMP4C may promote GC progression by negatively regulating necroptosis through inhibiting the phosphorylation of the RIPK1/RIPK3/MLKL signaling pathway,suggesting that it is expected to be a potential target for GC therapy.

AIM:This study aims to investigate the impact of chronic kidney disease(CKD)on the growth,development,and cognitive function of offspring rats,and to evaluate the effects of esaxerenone(ESAX)intervention.METHODS:Thirty female non-pregnant Wistar rats were randomly assigned to three groups:sham operation+pregnancy(control group),UUO+pregnancy(model group),and UUO+pregnancy+esaxerenone(treatment group),with 10 rats in each group.CKD was induced in the UUO+pregnancy and UUO+pregnancy+esaxerenone groups by performing uni-lateral ureteral obstruction(UUO),while the sham operation+pregnancy group underwent a non-ligated and non-clipped ureter procedure.Rats in the treatment group received esaxerenone at a dosage of 1 mg·kg-1·d-1.Vaginal smears were per-formed weekly from the 8th week post-UUO to monitor the estrous cycle.Female and male rats in pre-estrus or estrus were paired in a 2∶1 ratio,and the day with the first appearance of sperm in vaginal smears was recorded as the first day of preg-nancy.Offspring were delivered 21 to 22 days post-gestation,and 10 offspring from each group were selected for further experimentation:sham-offspring(sham-offspr)group,UUO-offspro group,and esaxerenone treatment offspring(ESAX-offspr)group.At 21 days of age,offspring weight and tail length were measured.Behavioral tests,including the open field test,Y maze test,and Morris water maze test,assessed learning and memory abilities.Serum levels of aldosterone,5-hydroxytryptamine(5-HT),and brain-derived neurotrophic factor(BDNF)were measured using ELISA.Serum creati-nine(SCr)and blood urea nitrogen(BUN)levels were assessed using conventional biochemical methods.Renal pathology was examined using Hematoxylin-Eosin(HE)staining.RESULTS:Offspring in the UUO-offspr group had reduced body weight and tail length compared to the sham-offspr group(P＜0.05).Behavioral tests indicated that exploration and memo-ry abilities were significantly impaired in the UUO-offspr group compared to the sham-offspr group(P＜0.05),while the ESAX-offspr group showed improved behavioral development compared to the UUO-offspr group(P＜0.05).ELISA results revealed decreased levels of serum 5-HT,BDNF,and aldosterone in the UUO-offspr group compared to the sham-offspr group(P＜0.01),with increased levels in the ESAX-offspr group(P＜0.05).Renal function tests showed elevated SCr and BUN levels in the UUO-offspr group compared to the sham-offspr group(P＜0.01),while levels in the ESAX-offspr group were reduced(P＜0.01).HE staining demonstrated mild tubular dilation and inflammatory cell infiltration in the UUO-offspr group,whereas the ESAX-offspr group had well-arranged tubules with reduced inflammation.CONCLU-SION:Chronic kidney disease adversely affects the growth,development,and cognitive function of offspring rats by 21 days of age.Esaxerenone intervention can mitigate these detrimental effects on growth,development,and cognitive abili-ties in offspring rats.

Objective:Exploring the role and mechanism of CHMP4C in regulating necroptosis during gastric can-cer development and progression.Method:The expression of CHMP4C in pan-cancer was analyzed by bioinformatics methods,and the expression of CHMP4C was detected in human normal gastric epithelial cells and GC cell lines by RT-qPCR and Western blot.Overexpression or knockdown of CHMP4C was performed in GC cell lines,and the effects of CHMP4C on the growth and proliferation of GC cells were detected using CCK-8 and clone formation assays.The CCK-8 experiment and Hoechst/PI double staining experiment were used to detect the changes in GC cell mortality and PI positive cell ratio after treatment with the necroptsis inducer TSZ or inhibitor necrostatin-1(Nec-1).Western blot assay was used to detect the protein and phosphorylation levels of RIPK1,RIPK3,and MLKL in GC cells.Result:CHMP4C was upregulated in GC tissues and cells.The CCK-8 and clone formation experiments showed that overex-pression of CHMP4C significantly improved the proliferation ability and colony formation efficiency of GC cells,while knockdown of CHMP4C significantly weakened GC cells.Moreover,the results of CCK-8 and Hoechst 33342/PI double staining experiments showed that upregulated CHMP4C could inhibit TSZ induced GC cell death;Nec-1 can reverse the decrease in GC cell viability caused by CHMP4C knockdown.Western blot experiment showed that the levels of p-RIPK1,p-RIPK3,and p-MLKL were significantly decreased in overexpressing cells,while they were increased in knockdown cells.After treatment with Nec-1,the expression levels of these three proteins decreased in knockdown cells.Conclusion:CHMP4C may promote GC progression by negatively regulating necroptosis through inhibiting the phosphorylation of the RIPK1/RIPK3/MLKL signaling pathway,suggesting that it is expected to be a potential target for GC therapy.

AIM:To investigate the possible mechanism how orphan nuclear receptor 4A1(NR4A1)affects renal fibrosis in unilateral ureteral obstruction(UUO)mice.METHODS:(1)Specific pathogen-free(SPF)male C57BL/6J mice(5 to 6 weeks old)were randomly divided into sh-Con+sham group(n=6),sham+sh-NR4A1 group(n=6),sh-NR4A1+UUO group(n=6),and sh-NR4A1+UUO group(n=6).Renal NR4A1 knockdown mice were prepared by intrarenal injection of NR4A1 viral vector.(2)SPF male C57BL/6J mice(5 to 6 weeks old)were randomly assigned to in sham group,UUO group,and UUO+cytosporone-B(Csn-B)group.An animal model of renal fibrosis was prepared by UUO,and Csn-B was intervened for 14 days.HE,Masson,and Sirius red staining were used to observe renal pathological damage.Immunohistochemistry and Western blot were performed to detect the expression of NR4A1,interferon-γ(IFN-γ),α-smooth muscle actin(α-SMA)and vimentin.The purpose of this study is to observe the regulatory effect of NR4A1 on UUO-induced renal fibrosis.RESULTS:(1)The expression of NR4A1 was decreased in kidney tissues of UUO mice(P<0.05).(2)HE staining results showed that there are tubular dilation,atrophy,and massive inflammatory cell infiltra-tion in sh-Con+UUO group,and NR4A1 knockdown can aggravate UUO-induced kidney damage(P<0.05).The results of Masson and Sirius red staining showed a banded distribution of collagen deposition in the sh-Con+UUO group,and colla-gen deposition increased significantly after NR4A1 knockdown(P<0.05).Treatment with Csn-B could improve renal path-ological damage and reduce collagen deposition.(3)UUO could upregulate the expression of α-SMA and vimentin,and NR4A1 knockdown could significantly increase UUO-induced their expression(P<0.05).In addition,Csn-B could im-prove UUO-induced renal fibrosis(P<0.05).(4)The expression of IFN-γ was increased in UUO mice,and NR4A1 knockdown could upregulate UUO-induced IFN-γ expression(P<0.05).Moreover,Csn-B could down-regulate UUO-in-duced IFN-γ expression(P<0.05).CONCLUSION:NR4A1 can affect renal fibrosis by regulating IFN-γ in UUO mice.

AIM:This study aims to investigate the impact of chronic kidney disease(CKD)on the growth,development,and cognitive function of offspring rats,and to evaluate the effects of esaxerenone(ESAX)intervention.METHODS:Thirty female non-pregnant Wistar rats were randomly assigned to three groups:sham operation+pregnancy(control group),UUO+pregnancy(model group),and UUO+pregnancy+esaxerenone(treatment group),with 10 rats in each group.CKD was induced in the UUO+pregnancy and UUO+pregnancy+esaxerenone groups by performing uni-lateral ureteral obstruction(UUO),while the sham operation+pregnancy group underwent a non-ligated and non-clipped ureter procedure.Rats in the treatment group received esaxerenone at a dosage of 1 mg·kg-1·d-1.Vaginal smears were per-formed weekly from the 8th week post-UUO to monitor the estrous cycle.Female and male rats in pre-estrus or estrus were paired in a 2∶1 ratio,and the day with the first appearance of sperm in vaginal smears was recorded as the first day of preg-nancy.Offspring were delivered 21 to 22 days post-gestation,and 10 offspring from each group were selected for further experimentation:sham-offspring(sham-offspr)group,UUO-offspro group,and esaxerenone treatment offspring(ESAX-offspr)group.At 21 days of age,offspring weight and tail length were measured.Behavioral tests,including the open field test,Y maze test,and Morris water maze test,assessed learning and memory abilities.Serum levels of aldosterone,5-hydroxytryptamine(5-HT),and brain-derived neurotrophic factor(BDNF)were measured using ELISA.Serum creati-nine(SCr)and blood urea nitrogen(BUN)levels were assessed using conventional biochemical methods.Renal pathology was examined using Hematoxylin-Eosin(HE)staining.RESULTS:Offspring in the UUO-offspr group had reduced body weight and tail length compared to the sham-offspr group(P＜0.05).Behavioral tests indicated that exploration and memo-ry abilities were significantly impaired in the UUO-offspr group compared to the sham-offspr group(P＜0.05),while the ESAX-offspr group showed improved behavioral development compared to the UUO-offspr group(P＜0.05).ELISA results revealed decreased levels of serum 5-HT,BDNF,and aldosterone in the UUO-offspr group compared to the sham-offspr group(P＜0.01),with increased levels in the ESAX-offspr group(P＜0.05).Renal function tests showed elevated SCr and BUN levels in the UUO-offspr group compared to the sham-offspr group(P＜0.01),while levels in the ESAX-offspr group were reduced(P＜0.01).HE staining demonstrated mild tubular dilation and inflammatory cell infiltration in the UUO-offspr group,whereas the ESAX-offspr group had well-arranged tubules with reduced inflammation.CONCLU-SION:Chronic kidney disease adversely affects the growth,development,and cognitive function of offspring rats by 21 days of age.Esaxerenone intervention can mitigate these detrimental effects on growth,development,and cognitive abili-ties in offspring rats.

Objective:To explore the clinical effects of free anterolateral thigh perforator flap (ALTPF) with modified cross-leg vessel bridging in reconstruction of infected wounds in the lower leg combined with major vessel defects.Methods:A retrospective observational study was conducted on 7 patients who admitted to the Department of Trauma Orthopaedics, the 521 Hospital of Norinco Group from January 2020 to December 2021 for treatment of large infected wounds in lower leg with soft tissue defect by reconstructive surgery of flap transfer. The patients were 5 males and 2 females, aged 23-50 years old with an average age of 37 years old. The causes of injury were: 5 patients were of car accidents, 1 of machinery compression and 1 of heavy object crush. The wounds were reconstructed after debridement and infection control with sensitive antibiotics, where the soft tissue defects were found at 11.0 cm×15.0 cm to 20.0 cm×32.0 cm in size. All patients underwent vascular angiography or CDU examinations and it was confirmed that the affected calf had only an anterior tibial artery as the vessel left for blood supply in 6 patients and a posterior tibial artery as the blood supply vessel in one patient. Therefore application of vascular end-to-side anastomosis in free flap reconstruction of limb defects was impossible due to the damaged artery could not be salvaged as a blood supply artery for the transferred flap. Therefore, a modified cross-leg vessel bridging to the freed ALTPF in the affected lower leg was applied. The donor site of the pedicle was covered with VSD while the pedicle of the flap was anastomosed. It was remained until the posterior tibial artery and the tubular flap were ready for replantation after disconnection of the pedicle. The sizes of flap were 13.0 cm×17.0 cm to 22.0 cm×32.0 cm (unilateral ALTPFs for 6 patients and bilateral ALTPFs for 1 patient). Two donor sites in low tension were direct closed, and the rest of 5 donor sites that had great tensions and could not be directly sutured were reconstructed by skin grafting. The survival and complications of flaps were observed in the scheduled postoperative follow-ups at outpatient visits, WeChat reviews and home visits, etc.Results:All 7 patients were successfully treated and had 12-24 months postoperative follow-up, with an average of 16 months. All flaps survived, with primary healing in 6 patients and 1 patient had partial flap necrosis with surface infection, which healed after dressing changes. The wound healing time was 14-36 days with an average of 17.9 days. The time for disconnection of the cross-leg vessel bridging pedicle was 3-4 weeks with the flap transfer, with an average of 3.6 weeks. The donor sites of ALTPFs and vessel pedicles all healed well. CDU confirmed the patency of the contralateral posterior tibial artery. Satisfactory functional recovery was achieved in the affected lower limb and there was a good function of the contralateral healthy lower leg.Conclusion:Application of the transfer of a free ALTPF with modified cross-leg vessel bridging in reconstruction of infected wounds with major vessel defects in the lower leg has shown excellent clinical outcomes. It is a practical and effective method in treatment of large infective defect in lower leg.

The main components and working principle of the fluid infusion and blood transfusion warmer were introduced,and the causes for the adverse events of the warmer were summarized based on National Medical Device Adverse Event Monitoring Information System and related literature in the world.The potential risks of the warmer were analyzed during operation,and the quality concerns and corresponding evaluation methods were proposed for the warmer from the aspects of device marking and documentation,structure,and temperature.References were provided for standard preparation,pre-market technical review and system verification of the fluid infusion and blood transfusion warmer.[Chinese Medical Equipment Journal,2023,44(10):76-80]

Objective:To investigate the clinical effect of gracilis musculocutaneous flap in repair of perineal soft defect with open pelvic fracture.Methods:From June 2009 to June 2019, 11 cases of open pelvic fracture associated with perineal injury were treated in the Department of Trauma and Orthopaedic of 521 Hospital of Norinco Group. There were 4 males and 7 females aged 16-56 (33 in average) years old. Cause of injuries: 6 cases by traffic accident, 4 by falling from height, and 1 by crushing. All the patients had open pelvic fractures. According to Tile classification, 1 case was rated as type A, 7 as type B and 3 as type C. All the patients were accompanied with perineal injury and soft tissue defect. The wound sizes ranged from 5 cm×5 cm to 8 cm×12 cm. The defects were repaired with gracilis musculocutaneous flap. The size of gracilis myocutaneous flaps was 6 cm×5 cm to 9 cm×13 cm. All donor areas of the flap were sutured directly. After surgery, 11 patients treated with strengthened nutritional support, keep supine position to avoid abduction, and appropriately raise the lower limbs. Follow-ups were conducted regularly after surgery.Results:All patients entered 6 to 30 (22 in average) months of follow-up. All of 11 myocutaneous flaps survived, besides 1 had a few necrosis at the distal surface of the myocutaneous flap, and healed after change of dressing. All the incisions at donor site had stage I healing. The colour, texture and flexibility of the gracilis myocutaneous flap were good. There was a scar at the donor sites without causing obvious dysfunction. Over the follow-up period, there was no failure of flap in either the recipient and donor sites. The patients were satisfied with the appearance and function.Conclusion:Gracilis musculocutaneous flap is one of the ideal methods in repair of perineal soft tissue defect with open pelvic fracture.

Objective:To evaluate the clinical efficacy and safety of Longmu Zhuanggu granule for the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency. Method:This multicenter stratified， block-randomized， double-blind， double-dummy， positive drug （pidotimod granule） parallel controlled， and non-inferiority trail intended to included 240 children patients and divided them into the experimental group （n=120） and the control group （n=120） at the ratio of 1∶1. Patients in both groups were treated for eight successive weeks and followed up for 12 months. The cure rates， numbers of respiratory infections， average courses of disease， curative effects of traditional Chinese medicine （TCM） syndrome， curative effects of individual symptoms， curative effects of immune indexes， and safety indexes between the two groups were observed and compared. Result:A total of 237 subjects were collected from 10 research centers， including 119 cases in the control group and 118 in the experimental group. There were 236 cases enrolled into the full analysis set （FAS）， 210 into the per-protocol set （PPS）， and 236 into the safety set （SS）. The baseline data of the two groups were not significantly different from each other， indicating that they were comparable. The cure rates of the experimental group and control group were 75.21% （88/117） and 73.95%（88/119）， respectively， with the 95% confidence interval （CI） of difference between the two groups being 1.26% （-9.85%，12.37%） for FAS and 3.81% （-6.28%，13.90%） for PPS. The 95% CI fell within the 10% non-inferiority margin， implying that non-infertility test of the cure rate in the treatment of endpoint disease was valid， and the conclusions of FAS and PPS analysis were consistent. There was no significant difference in the number or course of upper respiratory infection， bronchitis， and pneumonia. The difference in curative effects of TCM syndrome between the two groups after four weeks of treatment was not remarkable. After eight weeks of treatment， the total effective rate of the experimental group was 84.62%（99/117）， statistically higher than 78.15%（93/119） of the control group（χ²=-3.26，P<0.05）. There were no significant differences in the disappearance rates of individual symptoms between the two groups after four weeks of treatment. After eight weeks of treatment， the experimental group and control group exhibited the disappearance rates of 67.50%（54/80） and 47.37%（36/76） for shortness of breath and laziness to speak， 75.00%（54/72） and 53.33%（40/75） for poor appetite， 54.55%（60/110） and 37.84%（42/111） for hyperhidrosis， respectively， with obviously better outcomes observed in the experimental group （P<0.05，P<0.01）. The inter-group comparison revealed significant differences in immune indexes after eight weeks of treatment. As demonstrated by comparison with the situations before treatment， IgA， IgG， IgM， and CD4 did not change significantly after treatment. Except for CD8 in the experimental group (P<0.05), there was no significant difference in other immune indexes before and after treatment There was no significant difference in the incidence of adverse reactions. Conclusion:Longmu Zhuanggu granule is not inferior to pidomod granule in the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency， and it exhibits good safety， implying its promising clinical application value.