1.A prospective study of super-thin anterolateral thigh flap harvesting assisted by high-frequency color Doppler ultrasound in detecting perforators in deep adipose layers.
Qianyuan LIU ; Jiandong ZHOU ; Wencheng WANG ; Xueming CHEN ; Yajun XU ; Hai HUANG ; Jingyi MI
Chinese Journal of Reparative and Reconstructive Surgery 2024;38(1):62-68
		                        		
		                        			OBJECTIVE:
		                        			To investigate the clinical application of high-frequency color Doppler ultrasound (HFCDU) in detecting perforators in the deep adipose layers for harvesting super-thin anterolateral thigh flap (ALTF).
		                        		
		                        			METHODS:
		                        			Between August 2019 and January 2023, 45 patients (46 sides) with skin and soft tissue defects in the foot and ankle were treated, including 29 males and 16 females, aged from 22 to 62 years, with an average of 46.7 years. The body mass index ranged from 19.6 to 36.2 kg/m 2, with an average of 23.62 kg/m 2. The causes of injury included traffic accident injury in 15 cases, heavy object crush injury in 20 cases, mechanical injury in 8 cases, heat crush injury in 1 case, and chronic infection in 1 case. There were 20 cases on the left side, 24 cases on the right side, and 1 case on both sides. After thorough debridement, the wound size ranged from 5 cm×4 cm to 17 cm×11 cm. All patients underwent free super-thin ALTF transplantation repair. HFCDU was used to detect the location of the perforators piercing the deep and superficial fascia, as well as the direction and branches of the perforators within the deep adipose layers before operation. According to the preoperative HFCDU findings, the dimensions of the super-thin ALTF ranged from 6 cm×4 cm to 18 cm×12 cm. The donor sites of the flaps were directly sutured.
		                        		
		                        			RESULTS:
		                        			A total of 55 perforators were detected by HFCDU before operation, but 1 was not found during operation. During operation, a total of 56 perforators were found, and 2 perforators were not detected by HFCDU. The positive predictive value of HFCDU for identifying perforator vessels was 98.2%, and the sensitivity was 96.4%. Among the 54 perforators accurately located by HFCDU, the orientation of the perforators in the deep adipose layers was confirmed during operation. There were 21 perforators (38.9%) traveled laterally and inferiorly, 12 (22.2%) traveled medially and inferiorly, 14 (25.9%) traveled laterally and superiorly, 5 (9.3%) traveled medially and superiorly, and 2 (3.7%) ran almost vertically to the body surface. Among the 54 perforators accurately located by HFCDU, 35 were identified as type 1 perforators and 12 as type 2 perforators (HFCDU misidentified 7 type 2 perforators as type 1 perforators). The sensitivity of HFCDU in identifying type 1 perforators was 100%, with a positive predictive value of 83.3%. For type 2 perforators, the sensitivity was 63.2%, and the positive predictive value was 100%. The surgeries were successfully completed. The super-thin ALTF had a thickness ranging from 2 to 6 mm, with an average of 3.56 mm. All super-thin ALTF survived, however, 1 flap experienced a venous crisis at 1 day after operation, but it survived after emergency exploration and re-anastomosis of the veins; 1 flap developed venous crisis at 3 days after operation but survived after bleeding with several small incisions; 3 flaps had necrosis at the distal edge of the epidermis, which healed after undergoing dressing changes. All 45 patients were followed up 6-18 months (mean, 13.6 months). Three flaps required secondary defatting procedures, while the rest had the appropriate thickness, and the overall appearance was satisfactory.
		                        		
		                        			CONCLUSION
		                        			Preoperative application of HFCDU to detect the perforator in the deep adipose layers can improve the success and safety of the procedure by facilitating the harvest of super-thin ALTF.
		                        		
		                        		
		                        		
		                        			Male
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Thigh/surgery*
		                        			;
		                        		
		                        			Plastic Surgery Procedures
		                        			;
		                        		
		                        			Prospective Studies
		                        			;
		                        		
		                        			Skin Transplantation
		                        			;
		                        		
		                        			Free Tissue Flaps
		                        			;
		                        		
		                        			Burns
		                        			;
		                        		
		                        			Soft Tissue Injuries/surgery*
		                        			;
		                        		
		                        			Ultrasonography, Doppler, Color
		                        			;
		                        		
		                        			Crush Injuries/surgery*
		                        			;
		                        		
		                        			Perforator Flap
		                        			;
		                        		
		                        			Treatment Outcome
		                        			
		                        		
		                        	
2.Riluzole interferes with the activation of NLRP3 inflammasome in microglia of rats with spinal cord injury
Tao LIU ; Wenkai ZHANG ; Ziqian MA ; Yan ZHANG ; Xueming CHEN
Chinese Journal of Tissue Engineering Research 2024;28(7):1036-1042
		                        		
		                        			
		                        			BACKGROUND:Previous animal studies have shown that riluzole can inhibit neuroinflammatory response after spinal cord injury and promote functional recovery in injured rats,but the study on whether it can regulate the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome in the acute stage is lacking. OBJECTIVE:To observe whether riluzole can reduce microglial pyroptosis and promote functional recovery after spinal cord injury by modulating NLRP3 inflammasome through animal experiments,histological experiments and molecular biology experiments. METHODS:Female SD rats were divided into sham operation,model and riluzole groups,with 12 rats in each group.In addition to the sham operation group,T10 spinal cord injury was conducted in rats.The model group was treated with intraperitoneal administration of riluzole with solvent cyclodextrin.The riluzole group was treated with a 4 mg/kg dose of riluzole injection.The effect of riluzole on motor function recovery was assessed using the BBB score and inclined plane test.The recovery of sensory-evoked potential and motor-evoked potential was measured by electrophysiology.Hematoxylin-eosin staining was used to evaluate spinal cord tissue repair.The regulatory effects of riluzole on NLRP3,Caspase-1 and gasdermin D protein expression in spinal cord tissues were detected by western blot assay.ELISA was utilized to detect the expression levels of inflammatory factors interleukin-1β and interleukin-18.The effects of riluzole on the expression of NLRP3,Caspase-1,gasdermin D and interleukin-1β in microglial cells of the injured spinal cord were determined by immunofluorescence staining. RESULTS AND CONCLUSION:(1)At 35 days after spinal cord injury,BBB score and inclined plane test score in the riluzole group were higher than those in the model group(P<0.05).(2)At 3 days after spinal cord injury,the protein expressions of NLRP3,cleaved Caspase-1,gasdermin D-N(N-terminal domain),interleukin-1β,and interleukin-18 in the spinal cord homogenate of the riluzole group were significantly lower than those of the model group(P<0.05).(3)At 3 days after spinal cord injury,the fluorescence intensity of NLRP3,Caspase-1,gasdermin D and interleukin-1β in the riluzole group was significantly lower than that in the model group(P<0.05).(4)At day 35 after spinal cord injury,hematoxylin-eosin staining showed that the area of spinal cord injury in the riluzole group was smaller than that in the model group.Electrophysiological tests showed that the latency periods of sensory-evoked potential and motor-evoked potential in the riluzole group were shorter than those in the model group,and the latency period of wave amplitude in the riluzole group was higher than that in the model group.(5)These results suggest that riluzole can promote the repair of injured spinal cord tissue,promote the repair of nerve conduction function,and further promote the recovery of motor function in rats with spinal cord injury,which may be achieved through the regulation of NLRP3 inflammasome and the reduction of microglial pyroptosis.
		                        		
		                        		
		                        		
		                        	
3.Circ0005512 promotes microglia/macrophage pyroptosis after spinal cord injury in female rats
Yan ZHANG ; Wenkai ZHANG ; Wenxiu ZHANG ; Tao LIU ; Ziqian MA ; Xueming CHEN
Chinese Journal of Tissue Engineering Research 2024;28(31):5029-5035
		                        		
		                        			
		                        			BACKGROUND:Neuroinflammation is an important factor leading to secondary spinal cord injury,and microglia/macrophage pyroptosis is a significant part of post-spinal cord injury neuroinflammation.Studies have shown that microglia/macrophage undergoes pyroptosis after spinal cord injury,but the regulatory mechanism of circular RNA(circRNA)in microglia/macrophage pyroptosis after spinal cord injury remains unclear. OBJECTIVE:To investigate the role and mechanism of circRNA0005512 in regulating microglia/macrophage pyroptosis after spinal cord injury. METHODS:Female Wistar rats were divided into sham group and spinal cord injury group.Motor function was evaluated using the Basso,Beattie,and Bresnahan(BBB)scale.Cavity volume was assessed by hematoxylin-eosin staining.Differential expression of circRNA in spinal cord tissue was screened using RNA-sequencing and circ0005512 was validated by real-time PCR.Immunofluorescence,western blot assay,ELISA,and real-time PCR were performed to detect cell pyroptosis in the rats and lipopolysaccharide-induced microglial cell line HAPI cell models.Gene knockdown was used to confirm the regulatory role of circRNA0005512 in microglia/macrophage pyroptosis. RESULTS AND CONCLUSION:(1)Seven days after spinal cord injury,evident cavities were observed at the injury site.Immediately after spinal cord injury,the motor function of rats was completely lost.Over time,the motor function of rats in the spinal cord injury group gradually partially recovered,and there was a significant difference in BBB scores compared to the sham group.(2)Circ0005512 was significantly upregulated according to the results of the RNA-sequencing and confirmed in both the animal and cell models.(3)Immunofluorescence,western blot assay,real-time PCR,and ELISA confirmed the significant upregulation of cell pyroptosis markers(NLRP3,GSDMD,and caspase-1)in spinal cord injury tissue and lipopolysaccharide-induced HAPI cells.(4)In the cell model,knockdown of circ0005512 resulted in significantly decreased levels of cell pyroptosis marker-NLRP3.(5)The results above indicate that circ0005512 promotes pyroptosis in microglia/macrophages after spinal cord injury.
		                        		
		                        		
		                        		
		                        	
4.Single-centre diagnosis, treatment and prognostic analysis of abdominal aortic endograft infection
Xuebin WANG ; Bin LIU ; Zhe ZHANG ; Hongzhi YU ; Zhiwen ZHANG ; Lishan LIAN ; Xiang GAO ; Hai FENG ; Xueming CHEN
International Journal of Surgery 2024;51(11):759-765
		                        		
		                        			
		                        			Objective:To summarise and analyse the experience in the diagnosis and management of abdominal aortic endograft infection in recent years.Methods:Retrospectively summarised and analysed the general data, clinical presentation, laboratory and imaging findings, causative organisms and treatment choices of 14 patients with abdominal aortic endograft infection treated in Beijing Friendship Hospital, Capital Medical University, from January 2018 to June 2024, and analysed the prognosis of the patients and the risk factors associated with prognosis.Results:Positive bacterial cultures were 10 out of 14 patients. One non-operatively treated patient died of infectious toxic shock. Thirteen surgically treated patients underwent axillary-bifemoral artery bypass, removal of the infected stent, and closure of the aortic stump. Four of the 13 cases had combined aortoenteric fistula, 3 cases underwent one-stage enterocutaneous fistula repair, 1 case only fistula drainage, 3 cases of gastrojejunal anastomosis, all of them underwent gastric or jejunal nutrient tube implantation. Two of the 13 patients had combined the infection foci spread to the renal artery openings. To save the kidney, intraoperative left kidney autologous renal transplantation was performed in 1 case, and autologous saphenous vein reconstruction from celiac trunk artery-left renal artery and superior mesenteric artery-right renal artery was performed in the other case. All 14 patients were retrospectively summarised and followed up in August 2024, with 5 deaths in the early postoperative period (< 3 months), 3 deaths in the mid- to long-term period (≥3 months), and 5 survivors, with a median follow-up time of 2 years (1-5 years) for surviving patients. Among the 13 operated patients, 4 cases were combined with aortoenteric fistula, and 3 cases died in the early postoperative period; 4 cases of abdominal aortic infection foci involving renal artery openings, 2 cases of early postoperative death; 4 cases with pleural effusion, 4 cases died in the early postoperative period; 2 cases of combined creatinine elevation, 2 cases of early postoperative death; 2 cases of postoperative infection of artificial blood vessels.Conclusions:Abdominal aortic endograft infection are aggressive. The risk of early death is increased in patients who are elderly, in poor general condition, with aortoenteric fistula or with pre-existing cardiac, pulmonary, hepatic and renal insufficiency, but surgery based on adequate anti-infective therapy remains an effective means of saving the patient′s life.
		                        		
		                        		
		                        		
		                        	
5.Comparison of clinical and injured vertebra radiological parameters between patients with non-traumatic osteoporotic vertebral compression fracture and those with traumatic one after percutaneous vertebroplasty
Benqiang TANG ; Xueming CHEN ; Libin CUI ; Yanhui WANG ; Xin YUAN ; Yadong LIU ; Peng ZHAO ; Liang LIU
Chinese Journal of Orthopaedic Trauma 2024;26(11):956-963
		                        		
		                        			
		                        			Objective:To explore the differences in clinical and injured vertebra radiological parameters between patients with non-traumatic osteoporotic vertebral compression fracture (OVCF) and those with traumatic OVCF after percutaneous vertebroplasty (PVP).Methods:A retrospective study was conducted to analyze the 369 OVCF patients (with 458 vertebrae injured) who had been treated by PVP at Department of Orthopaedics, Beijing Luhe Hospital between October 2015 and March 2017. There were 292 females and 77 males with a mean age of 73 (60, 79) years. Based on the absence or presence of a trauma history, the patients were stratified into a non-traumatic group (127 patients with 160 vertebrae injured) and a traumatic group (242 patients with 298 vertebrae injured). Clinical parameters [age, gender, body mass index, symptomatic duration, and number of injured vertebrae, visual analogue scale (VAS), Oswestry disability index (ODI), duration of follow-up, and rate of new OVCFs] and injured vertebra radiological parameters (position of injured vertebra, fracture type, compression severity, fracture range, cortical defect, intravertebral cleft, spinal canal compromise, basivertebral foramen, morphology of bone cement, range of bone cement, cement leakage, cement volume, rate of vertebral height restoration, recollapse of cemented vertebrae) were recorded perioperatively. All the clinical and radiological parameters were compared between the 2 groups.Results:Compared with the traumatic group, the non-traumatic group had an older age [75 (71, 83) years versus 71 (65, 76) years], more females (85.0% versus 76.0%), a longer symptomatic duration [10.0 (7.0, 15.0) d versus 6.5 (2.0, 12.0) d], a lower preoperative VAS pain score [7 (6, 8) points versus 7 (7, 8) points], a lower VAS pain score at postoperative day 1 [2 (2, 3) points versus 2 (2, 3) points], a lower preoperative ODI [66% (63%, 72%) versus 70% (65%, 73 %)], a lower ODI at postoperative day 1 [32% (30%, 34%) versus 32% (31%, 34%)], a higher rate of new OVCFs during follow-up (34.6% versus 12.8%), a lower rate of thoracolumbar lesions (51.9% versus 70.1%), more deformed fractures (mostly amphicoelous type), a lower rate of cortical defects in the anterior wall (20.0% versus 31.5%), a higher rate of trabecular pattern of cement (83.1% versus 71.8%), a higher rate of type-B cement leakage (50.6% versus 31.9%), a lower rate of type-C cement leakage (5.6% versus 12.8%), a lower rate of recollapse of cemented vertebrae (43.8% versus 55.4%). All the comparisons above were statistically significant ( P<0.05). There were no significant differences between the 2 groups in the other clinical or radiological parameters ( P>0.05). Conclusions:There are statistically significant differences in a significant number of clinical and injured vertebra radiological parameters between patients with non-traumatic OVCF and those with traumatic OVCF after PVP. It is noteworthy that non-traumatic OVCFs are one specific subgroup of OVCFs.
		                        		
		                        		
		                        		
		                        	
6.Microbiomes combined with metabolomics reveals the changes of microbial and metabolic profile of articular cavity effusion in rheumatoid arthritis, urarthritis and osteoarthritis patients
Hanzhi Yi ; Wukai Ma ; Minhui Wang ; Chunxia Huang ; Guangzhao Gu ; Dan Zhu ; Hufan Li ; Can Liu ; Fang Tang ; Xueming Yao ; Liping Sun ; Nan Wang ; Changming Chen
Acta Universitatis Medicinalis Anhui 2024;59(12):2237-2245
		                        		
		                        			Objective:
		                        			To investigate the changes of microorganisms and metabolites in joint effusion of patients with Rheumatoid arthritis(RA), Osteoarthritis(OA) and Urarthritis(UA). To provide new ideas for the study of the effect of microbiota on the pathogenesis of arthritis. 
		                        		
		                        			Methods:
		                        			Joint effusion samples were collected from 20 patients with RA, 20 patients with OA, and 20 patients with UA. 16S rRNA gene sequencing and untargeted ultra-high performance Liquid chromatography-mass spectrometry(LC-MS) were used to explore the differences in microorganisms and metabolites among the three groups. Pearson correlation analysis was used to detect the correlation between effusion microbiota and metabolites.
		                        		
		                        			Results:
		                        			There were differences in microbial diversity and microbiota composition among the three groups. Combined with VIP>1 from OPLS-DA andP<0.05 from two-tailed Students t-test, 45 differential metabolites(Between RA and OA groups), 38 differential metabolites(Between UA and OA groups) and 16 differential metabolites(Between RA and UA groups), were identified. GO analysis and KEGG pathway analysis showed that the differential metabolic pathways among the three groups were mainly concentrated in citric acid cycle(TCA cycle), nucleotide metabolism, amino acid metabolism and glycolysis pathway. Correlation analysis of joint effusion microbiota and metabolites suggested that bacteria enriched in the three groups of joint effusion, such asPrevotella,Clostridium ruminosus,Prevotellaceae_UCG-001, were related to many key metabolites such as lysozyme, uric acid, glucose, and L-glutamine.
		                        		
		                        			Conclusion
		                        			This study shows that there are a variety of bacterial flora in joint cavity effusion of RA, OA, and UA patients, and the differential metabolites produced by them are involved in the pathogenesis of the three types of arthritis by affecting a variety of metabolic pathways.
		                        		
		                        		
		                        		
		                        	
7.Anterolateral thigh Flow-through flap transfer combined with Masquelet technique: in emergency surgery of limb salvage for Gustilo IIIC distal tibial fractures
Xueguang LIU ; Jiandong ZHOU ; Zheng CHEN ; Xingfei ZHANG ; Tonglong XU ; Xueming CHEN ; Yajun XU
Chinese Journal of Microsurgery 2024;47(3):261-266
		                        		
		                        			
		                        			Objective:To investigate the therapeutic effect of anterolateral thigh Flow-through flap transfer surgery combined with Masquelet technique in reconstruction of Gustilo IIIC open fractures of distal tibia.Methods:Between July 2017 and May 2021, 7 patients who had Gustilo IIIC injuries in the lower limb were treated in the Department of Orthopaedics Surgery, Wuxi 9th People's Hospital by emergency surgery with transfer of anterolateral thigh Flow-through flap combined with Masquelet technique. The patients were 5 males and 2 females, aged 36 to 63 (50.0±10.4) years old. Size of soft tissue defects was 11 cm × 4 cm to 23 cm × 7 cm, the length of bone defects was 3.5-7.5 (5.34±1.52) cm and the bridging length for vascular defects was 7-12 (9.21 ± 2.34) cm. The size of the flaps was 12 cm × 5 cm - 24 cm × 8 cm. All patients received postoperative follow-up at the outpatient clinic and complications of wound, fracture healing and the recovery of limb function were observed.Results:All flaps survived uneventfully and successful limb salvage were achieved in all 7 patients, together with all bone grafts healed without infection. The follow-up lasted for 12-38 (26.69±10.73) months. At the last follow-up, the appearance and functional recovery of the lower limbs were satisfactorily. The function of ankle was evaluated according to American Orthopedic Foot and Ankle Society (AOFAS) : 3 patients in excellent, 3 in good and 1 in fair.Conclusion:Emergency anterolateral thigh Flow-through flap transfer surgery with Masquelet technique is a safe, effective and feasible surgical procedure for Gustilo IIIC open fractures of distal tibia. It allows to close the wound and rebuild the blood supply in distal limb in the primary or emergency surgery, and allows to perform bone grafting and internal fixation in stage-II surgery. The patients benefit from high rate of success in limb salvage and good function recovery of the affected limb.
		                        		
		                        		
		                        		
		                        	
8.Mechanical thrombus aspiration combined with superior mesenteric artery transcatheter thrombolysis in the treatment of portal vein thrombosis
Bin LIU ; Hai FENG ; Xueming CHEN ; Wenrui LI ; Hongzhi YU
International Journal of Surgery 2023;50(3):155-159,F3
		                        		
		                        			
		                        			Objective:To compare the effectiveness and safety of mechanical thrombus aspiration combined with superior mesenteric artery transcatheter thrombolysis with those of simple superior mesenteric artery transcatheter thrombolysis in the treatment of portal vein-superior mesenteric vein thrombosis.Methods:A retrospective case-control study was used to analyze the clinical data of 27 patients with portal vein thrombosis treated by interventional therapy in Beijing Friendship Hospital Affiliated to Capital Medical University from February 2020 to October 2022. According to different interventional procedures, they were divided into two groups: 13 cases were treated with mechanical thrombus aspiration combined with superior mesenteric artery catheterization thrombolysis (combined treatment group), and 14 cases were treated with superior mesenteric artery catheterization thrombolysis alone (catheterization thrombolysis group). The postoperative and preoperative portal vein thrombus grade, catheterization thrombolysis time, parenteral nutrition time and operation-related complications were observed and compared between the two methods, and the incidence of long-term intestinal necrosis was compared after postoperative follow-up. Measurement data of normal distribution were expressed as mean±standard deviation( ± s), t-test was used for comparison between groups. Counting data were expressed as cases and percentage(%), and comparison between groups was used Chi-square test or Fisher exact probability method. Results:There was significant difference in the grade of portal vein thrombosis between the combined treatment group and the catheterization thrombolysis group( P<0.05). The thrombolytic time of catheterization in the two groups was (2.38±0.74) d and (4.79±1.15) d, respectively, and the time of parenteral nutrition was (4.08±2.87) d and (8.50±3.16) d, respectively. The difference was statistically significant ( P<0.05). There was no significant difference in operation-related complications between the two groups ( P>0.05). There were no severe injury of liver and kidney function in both groups. One patient in each group underwent long-term enterectomy. Conclusions:Mechanical thrombus aspiration combined with superior mesenteric artery thrombolysis is safe and effective in the treatment of portal vein thrombosis. Compared with simple superior mesenteric artery thrombolysis, mechanical thrombus aspiration combined with superior mesenteric artery thrombolysis can increase the rate of thrombus clearance and reduce the time of thrombolysis and parenteral nutrition support.
		                        		
		                        		
		                        		
		                        	
9.Clinical application of G-iliac? iliac branch device in the treatment of common iliac artery aneurysm
Wenrui LI ; Hai FENG ; Zhiwen ZHANG ; Bin LIU ; Hongzhi YU ; Xiang GAO ; Zhe ZHANG ; Xueming CHEN
International Journal of Surgery 2023;50(7):464-468,C2
		                        		
		                        			
		                        			Objective:To analyze the safety and efficacy of G-iliac? iliac branch device (IBD) in the treatment of common iliac artery aneurysm.Methods:The clinical data of 7 patients with common iliac artery aneurysm who were treated with G-iliac? IBD and internal iliac artery (IIA) preserved were retrospectively analyzed in the Department of Vascular Surgery, Beijing Friendship Hospital, Capital Medical University from June 2021 to June 2022, and the surgical effects and related complications were analyzed.Results:All 7 patients were male, aged from 57 to 80 years, with an average age of 70.9 years. There were 6 cases of abdominal aortic aneurysm combined with common iliac artery aneurysm and 1 case of simple common iliac artery aneurysm, all of them were successfully applied with G-iliac? IBD to preserve IIA. Cardiogenic shock occurred in 1 patient after the operation. 7 patients were followed up for 3-15 months, with an average of 8 months. During the follow-up period, the iliac artery and IIA stents were all patency, and there was no IBD-related endoleak, stent displacement, buttock claudication, sexual dysfunction, or aortic-related death. The diameter of abdominal aortic aneurysm and common iliac artery aneurysm were stable.Conclusion:For patients with common iliac artery aneurysm, preservation of IIA with G-iliac? IBD is a safe and effective technique with a high technical success rate and IIA patency rate, and has a low complication rate, but the long-term effect still requires more data and longer follow-up data to support.
		                        		
		                        		
		                        		
		                        	
10.Effect of Cuscutae Semen on Learning and Memory and N6-methyladenosine Modification in Hippocampus of Offspring of Fear-damaged Pregnant Rats
Tong ZHANG ; Junlin HOU ; Yu JIANG ; Liping YANG ; Zhixin DU ; Yuexuan LIU ; Shenao DING ; Xueming YANG ; Yinan YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(2):104-112
		                        		
		                        			
		                        			ObjectiveTo observe the effects of Cuscutae Semen on the learning and memory ability, N6-methyladenosine (m6A)-related modification enzymes and total m6A level in hippocampus of the offspring of fear-damaged pregnant rats. MethodForty-five pregnant rats were randomly divided into blank group, model group and Cuscutae Semen group. From the 1st day to the 19th day of pregnancy, rats in the model group and the Cuscutae Semen group were induced by observing electric shock of other rats. The Cuscutae Semen group was treated with 5 g·kg-1·d-1 Cuscutae Semen decoction (ig), while the other two groups were treated with the same amount of purified water. The offspring were assigned following the grouping method of their maternal generation. The behavioral changes of the offspring were tested by Morris water maze on 21st day after birth, and the development of hippocampal neurons was observed by transmission electron microscopy. The mRNA and protein expression levels of methyltransferase-like 3 (METTL3), METTL14, Wilms tumor 1 associated protein (WTAP), fat mass and obesity-associated protein (FTO) and Alk B homologue 5 (ALKBH5) were detected by Real-time polymerase chain reaction (Real-time PCR), Western blot and immunohistochemistry (IHC). The total content of m6A in hippocampus was determined by high performance liquid chromatography tandem mass spectrometry (LC-MS/MS). ResultCompared with the conditions in the blank group, the average latency duration in the model group was prolonged, and the number of entries in the target quadrant, the target quadrant duration and the number of crossing the platform were decreased (P<0.01). Additionally, the model group had seriously damaged structure of hippocampal CA1 and CA3 neurons, swollen mitochondria, expanded endoplasmic reticulum, and small number of synapses with some having blurred structure, and the expression levels of METTL3, METTL14, FTO, ALKBH5 as well as the total m6A level were lower than those in the blank group (P<0.05,P<0.01). Compared with the model group, the Cuscutae Semen group had shortened average latency duration, increased number of entries in the target quadrant, target quadrant duration and number of crossing the platform (P<0.01), alleviated damage of hippocampal CA1 and CA3 neurons, fine structure of mitochondrial and endoplasmic reticulum, and clear, intact and dense synapses. And the expression levels of METTL3, METTL14, FTO as well as the total level of m6A were up regulated, while the expression level of ALKBH5 was down regulated in the Cuscutae Semen group (P<0.05,P<0.01). ConclusionCuscutae Semen improved the learning and memory ability of the offspring of the rats affected by fear damaging kidney during pregnancy, protected hippocampal neurons, and up-regulated the expression levels of METTL3, METTL14, FTO and the total m6A level in hippocampus. 
		                        		
		                        		
		                        		
		                        	
            

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