1.The long-term follow-up of emergency repair of partial auricle of complete separation by superficial temporal fascia flap combined with severed auricle reimplantation
Chen CHEN ; Zhenyu WANG ; Hongyi WANG ; Yuan JIN ; Jiulong LIANG
Chinese Journal of Plastic Surgery 2024;40(8):878-884
Objective:To observe the long-term effect of superficial temporal fascia flap combined with severed auricle reimplantation in emergency repair of partial auricle of complete separation.Methods:The data of patients with partial auricle of complete separation admitted to Emergency Clinic of Burn and Plastic Surgery of General Hospital of Northern Theater Command from June 2014 to August 2023 were retrospectively analyzed. All of them were repaired with a superficial temporal fascia flap combined with amputating auricle reimplantation. During the operation, the superficial temporal fascia flap was harvested, and the pedicle was preserved. Then the detached auricular cartilage was removed and used as a replantation scaffold. Then the remaining skin was thinned to create a full-thickness skin graft after cartilage detachment. Referring to the position and angle of the contralateral auricle, the cartilage scaffold was sutured and fixed at the stump of the ear cartilage. The wound was covered with a superficial temporal fascia flap and a full-thickness skin graft, and then packed and sutured. The postoperative observation indicators mainly confirm whether the surgery was successful, the healing condition of the replanted ear, and whether there were problems such as skin flap necrosis, infection, hematoma, etc. The shape, color, texture and tactile recovery of the reconstructed auricle were evaluated by long-term follow-up for more than 1 year. The Vancouver scar scale (VSS) was used to assess scarring in both donor and recipient sites (total score of 0-15 points, higher scores indicated more severe scarring). The Likert 5-level scoring method was used to evaluate the patients’ satisfaction with the surgical results (total score of 30 points, ≥27 points were very satisfied, 24-26 points were somewhat satisfied, 18-23 points were indifferent, 15-17 points were somewhat dissatisfied, ≤14 points were very dissatisfied).Results:A total of 8 patients were enrolled, including 5 males and 3 females. Their ages ranged from 26 to 65 years, with an average of 41 years. All patients had unilateral ear defects, with 3 cases in the left ear and 5 cases in the right ear. The defect areas ranged from 1.5 cm × 2.5 cm to 5.0 cm × 4.0 cm. During the surgery, the harvested superficial temporoparietal fascia flaps ranged from 4.5 cm × 6.5 cm to 15.0 cm × 10.0 cm. After surgery, both the flaps and full-thickness skin graft healed satisfactorily, with primary healing observed in both the donor and recipient sites. There were no complications such as necrosis, infection, or hematoma were observed. The follow-up period ranged from 3 to 9 years, with an average of approximately 6.3 years. Except for one case that required a defatted surgery at the second stage, the reconstructed auricles of the remaining patients were basically consistent with the healthy side, with smooth contour lines, skin color and texture close to the surrounding tissues, and improved tactile sensitivity. In the final follow-up, the VSS scores for both the donor and recipient sites were ≤3 for all patients. All patients rated the surgical outcome as very satisfied.Conclusion:For the partial auricle of complete separation that has no chance of replantation, the use of superficial temporal fascia flap and detached ear composite graft to repair is a reliable and effective surgical method. Patients have good postoperative long-term effects and high levels of satisfaction.
2.The long-term follow-up of emergency repair of partial auricle of complete separation by superficial temporal fascia flap combined with severed auricle reimplantation
Chen CHEN ; Zhenyu WANG ; Hongyi WANG ; Yuan JIN ; Jiulong LIANG
Chinese Journal of Plastic Surgery 2024;40(8):878-884
Objective:To observe the long-term effect of superficial temporal fascia flap combined with severed auricle reimplantation in emergency repair of partial auricle of complete separation.Methods:The data of patients with partial auricle of complete separation admitted to Emergency Clinic of Burn and Plastic Surgery of General Hospital of Northern Theater Command from June 2014 to August 2023 were retrospectively analyzed. All of them were repaired with a superficial temporal fascia flap combined with amputating auricle reimplantation. During the operation, the superficial temporal fascia flap was harvested, and the pedicle was preserved. Then the detached auricular cartilage was removed and used as a replantation scaffold. Then the remaining skin was thinned to create a full-thickness skin graft after cartilage detachment. Referring to the position and angle of the contralateral auricle, the cartilage scaffold was sutured and fixed at the stump of the ear cartilage. The wound was covered with a superficial temporal fascia flap and a full-thickness skin graft, and then packed and sutured. The postoperative observation indicators mainly confirm whether the surgery was successful, the healing condition of the replanted ear, and whether there were problems such as skin flap necrosis, infection, hematoma, etc. The shape, color, texture and tactile recovery of the reconstructed auricle were evaluated by long-term follow-up for more than 1 year. The Vancouver scar scale (VSS) was used to assess scarring in both donor and recipient sites (total score of 0-15 points, higher scores indicated more severe scarring). The Likert 5-level scoring method was used to evaluate the patients’ satisfaction with the surgical results (total score of 30 points, ≥27 points were very satisfied, 24-26 points were somewhat satisfied, 18-23 points were indifferent, 15-17 points were somewhat dissatisfied, ≤14 points were very dissatisfied).Results:A total of 8 patients were enrolled, including 5 males and 3 females. Their ages ranged from 26 to 65 years, with an average of 41 years. All patients had unilateral ear defects, with 3 cases in the left ear and 5 cases in the right ear. The defect areas ranged from 1.5 cm × 2.5 cm to 5.0 cm × 4.0 cm. During the surgery, the harvested superficial temporoparietal fascia flaps ranged from 4.5 cm × 6.5 cm to 15.0 cm × 10.0 cm. After surgery, both the flaps and full-thickness skin graft healed satisfactorily, with primary healing observed in both the donor and recipient sites. There were no complications such as necrosis, infection, or hematoma were observed. The follow-up period ranged from 3 to 9 years, with an average of approximately 6.3 years. Except for one case that required a defatted surgery at the second stage, the reconstructed auricles of the remaining patients were basically consistent with the healthy side, with smooth contour lines, skin color and texture close to the surrounding tissues, and improved tactile sensitivity. In the final follow-up, the VSS scores for both the donor and recipient sites were ≤3 for all patients. All patients rated the surgical outcome as very satisfied.Conclusion:For the partial auricle of complete separation that has no chance of replantation, the use of superficial temporal fascia flap and detached ear composite graft to repair is a reliable and effective surgical method. Patients have good postoperative long-term effects and high levels of satisfaction.
3.Real-world Study of the Safety and Efficacy of Pembrolizumab in the Treatment of Advanced Non-small Cell Lung Cancer
WAN NING ; WANG BING ; GUO YA ; HE ZIJIAN ; YANG CHEN ; YANG NING ; LU LIQING ; LIANG HONGYI ; XIAO WEIBIN ; YANG DANDAN ; CHEN ZHUOJIA ; FANG WENFENG ; LIANG WEITING
Chinese Journal of Lung Cancer 2024;27(10):745-754
Background and objective Pembrolizumab(PEM)has been shown to be effective in clinical trials for the treatment of advanced non-small cell lung cancer(NSCLC),but clinical trials were based on cohorts of patients selected on specific criteria,and whether the findings are consistent with real-world patients is debatable.The aim of this study is to evaluate the efficacy and safety of PEM in the treatment of advanced NSCLC based on real-world data.Methods A retro-spective collection of real-world data from patients with advanced NSCLC receiving PEM was conducted.Propensity score matching was used to eliminate inter-group differences and assess the efficacy and safety of PEM compared to chemotherapy.Results Among 450 matched patients,the incidence rates of any-grade adverse events were 79.87%in the PEM group and86.71%inthe chemotherapy group,while the incidence rates of grade>3 adverse events were 4.03%and 7.31%,respectively.The objective response rates were 48.63%for PEM and 36.00%for chemotherapy(P=0.011).The median progression-free survival was 15.5 months for PEM and 8.8 months for chemotherapy(P<0.001),and the median overall survival was not reached for PEM and 26.2 months for chemotherapy(P<0.001).Conclusion PEM treatment for advanced NSCLC demonstrates favorable survival outcomes and acceptable safety in real-world clinical practice.
4.Treatment of infected nonunion after internal fixation of subtrochanteric fracture with a reconstruction stent of external fixation
Yonghui FAN ; Lei HUANG ; Zhilin XIA ; Weidong MING ; Jianfeng LI ; Jianfeng PEI ; Hongyi YAO ; Jiebin DUAN ; Kangxiong LIANG
Chinese Journal of Orthopaedic Trauma 2023;25(4):310-318
Objective:To evaluate the treatment of infected nonunion after internal fixation of subtrochanteric fracture with a reconstruction stent of external fixation.Methods:A retrospective study was conducted to analyze the data of 5 male patients with infected nonunion after internal fixation of subtrochanteric fracture who had been treated and completely followed up at The Great Wall Orthopaedics and Hand Surgery Hospital from January 2017 to October 2022. The patients were (30.0±13.5) years old. Seinsheimer fracture types: ⅢA (1 case), ⅢB (1 case), Ⅳ (2 cases), and Ⅴ (1 case); original internal fixation: intramedullary system (4 cases) and plate fixation (1 case); the Cierny-Mader anatomical classification: type Ⅳ (diffuse type) for all. After complete debridement at stage one, 2 or 3 hydroxyapatite (HA) coated screws were placed at both fracture ends from the lateral side of the femur for unilateral reconstruction external fixation. Next, a hybrid external fixation scaffold was added with a 1/3 ring at the sagittal position and 1 or 2 HA screws in 4 cases while unilateral reconstruction external fixation was constructed at both sides by inserting 2 HA screws into both fracture ends from the anterior femur at the sagittal position in 1 case. Antibiotic bone cement was used to fill bone defects of (3.8±1.8) cm. At 6 to 8 weeks after debridement when infection did not recur, antibiotic bone cement was removed before autogenous iliac bone grafting was performed in 3 patients and osteotomy bone transport in 2 patients. Infection control, bone union time, time for removal of external fixation stent, complications, Sanders hip function score and Paley bone outcome score were recorded.Results:The 5 patients were followed up for (23.4±8.1) months after surgery. Infection at the fracture ends was controlled after 1 time of debridement in 3 patients and after 2 times of debridement in 2 patients. The loosening HA screws were replaced twice due to infection at the proximal nail tract, and autologous bone grafting was performed at the opposite fracture ends in 1 case; no complications occurred in the other 4 cases. Bony union was achieved at the extended segment and fracture ends in all patients. The time for imaging union after bone reconstruction was (10.2±3.4) months. The time for wearing a stent of external fixation was (18.0±4.5) months. There was no recurrent infection or lingering infection. According to the Sanders hip function score at the last follow-up, 4 cases were excellent and 1 case was good; according to the Paley bone outcome score, the curative effect was excellent in all.Conclusion:Application of a reconstruction stent of external fixation combined with antibiotic bone cement can control infection at the first stage and conduct bone reconstruction at the second stage to successfully treat the infected nonunion and preserve the hip function after internal fixation of subtrochanteric fracture.
5.Effects of modified proper digital artery island flap in repairing complex fingertip defects
Yue HUANG ; Feng LIN ; Chunyu HAO ; Jiulong LIANG ; Dapeng ZHOU ; Hongyi WANG
Chinese Journal of Burns 2023;39(10):947-952
Objective:To investigate the surgical method and clinical effects of the modified proper digital artery island flap in repairing complex fingertip defects.Methods:A retrospective observational study was conducted. From January 2017 to December 2021, 15 patients (15 fingers) with complex fingertip defects, involving the pulp, nail bed, and lateral wall of the nail, who met the inclusion criteria were admitted into General Hospital of Northern Theater Command, including 11 males and 4 females, aged from 18 to 55 years. The area of the post debridement wound was from 2.5 cm×2.0 cm to 3.5 cm×3.5 cm, and all the wounds were repaired by using modified proper digital artery island flap (including 3 parts: main flap, tongue-shaped flap, and triangular flap), of which the main flap was used to cover the finger pulp defect, the tongue-shaped flap was used to cover the nail bed and the nail lateral wall defect, and the triangular flap was inserted into the edge of the finger pulp wound to cover the vessel pedicle. The range of the flap ranged from 3.0 cm×2.0 cm to 4.5 cm×3.0 cm. The wound at the donor site was repaired with full-thickness skin graft of the groin, and the donor site of the skin graft was sutured directly. After operation, the survival of the flap and skin graft as well as and the appearance of the affected finger were observed. During the follow-up, the fingertip morphology of the affected finger was observed, two-point discrimination distance of the affected finger pulp was measured, and the patients' satisfaction with the efficacy (including very satisfied, satisfied, and dissatisfied) was asked, and the affected finger function was evaluated by the total active movement (TAM) system evaluation standard recommended by American Academy for Surgery of Hand.Results:After operation, the main flaps and skin grafts in 15 patients all survived; but the incision at the edge of tongue-shaped flap in one patient healed poorly, and one patient developed venous stasis at the distal end of the tongue-shaped flap; the triangular flap at the pedicle was slightly bloated in the early postoperative period and became smooth after 2 to 3 months. Overall, two patients developed subcutaneous hematoma in their flaps. All the complications were healed by appropriate dressing change, suture removal, or compression bandaging. After operation, the appearance of the flap was full and formed a prominent fingertip shape. During the follow-up of 6 months to 5 years, the fingertips of the affected fingers were prominent and full; the two-point discrimination distance of the affected finger pulp was (8.6±1.4) mm; 8 patients were very satisfied with the efficacy, 6 patients were satisfied, and one patient was dissatisfied; the functional assessment of the affected fingers were all excellent.Conclusions:The modified proper digital artery island flap can repair complex fingertip defects involving the pulp, nail bed, and lateral wall of the nail. The operation is simple, and the shape and function of the fingertip are good after surgery.
6.Effects of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique
Zhenyu WANG ; Hongyi WANG ; Yuan JIN ; Tinghui ZHANG ; Zhishan XU ; Jiulong LIANG
Chinese Journal of Plastic Surgery 2023;39(11):1222-1228
Objective:To observe the efficacy the application of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia.Method:In this retrospective study, the clinical data were collected from the patients who received microtia reconstruction with autologous rib cartilage at the Department of Burns and Plastic Surgery, General Hospital of Northern Theater Command from January 2015 to January 2022. According to the surgical procedure, patients were divided into endoscopic group and open surgery group. In endoscopic group, endoscope-assisted temporoparietal fascia harvest were performed for the second-stage operation in auricular reconstruction of Nagata’s technique. In open surgery group, temporoparietal fascia flaps were harvested in open surgery for the second-stage operation in auricular reconstruction of Nagata’s technique. Regular follow-up was conducted to observe the survival of the fascia flaps, complications, patient satisfaction, and surgical scars. The patient satisfaction questionnaire for auricular reconstruction was used to assess patient satisfaction, and the patient and observer scar assessment scale (POSAS) was used to evaluate scar formation in the surgical area. Data analysis was performed using SPSS 26.0 statistical software. The measurement data were expressed by Mean ± SD, and the counting data were expressed as cases (%). The T-test was used to compare the age difference, length of hospital stay, intraoperative blood loss, scar length, patient satisfaction, and POSAS scores between the two groups. Chi-square test was used to compare the gender composition and incidence of complications between the two groups. P<0.05 was considered statistically significant. Results:A total of 51 patients were included, with 26 in the endoscopic group (14 men and 12 women) and 25 in the open surgery group (12 men and 13 women). The age of the patients in the endoscopic group was (9.8±2.9) years (ranging from 7 to 17 years), while in the open surgery group was (10.3±3.8) years (ranging from 7 to 17 years). The postoperative follow-up period was (15.4±3.4) months (1 to 2 years), and all fascia flaps survived without any severe complications. There were no statistically significant differences between the two groups in terms of age difference, length of hospital stay, intraoperative blood loss, postoperative satisfaction, sex composition ratio, and postoperative complications ( P>0.05). The scar quality in the endoscopy group was superior to that in the open surgery group, and POSAS scores of endoscopic group were lower than those in the open surgery group, and the difference was statistically significant ( P<0.05). Conclusion:Endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia can minimize scarring, improve the postoperative appearance and is not statistically associated with the appearance of reconstructed auricles or complications.
7.Effects of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique
Zhenyu WANG ; Hongyi WANG ; Yuan JIN ; Tinghui ZHANG ; Zhishan XU ; Jiulong LIANG
Chinese Journal of Plastic Surgery 2023;39(11):1222-1228
Objective:To observe the efficacy the application of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia.Method:In this retrospective study, the clinical data were collected from the patients who received microtia reconstruction with autologous rib cartilage at the Department of Burns and Plastic Surgery, General Hospital of Northern Theater Command from January 2015 to January 2022. According to the surgical procedure, patients were divided into endoscopic group and open surgery group. In endoscopic group, endoscope-assisted temporoparietal fascia harvest were performed for the second-stage operation in auricular reconstruction of Nagata’s technique. In open surgery group, temporoparietal fascia flaps were harvested in open surgery for the second-stage operation in auricular reconstruction of Nagata’s technique. Regular follow-up was conducted to observe the survival of the fascia flaps, complications, patient satisfaction, and surgical scars. The patient satisfaction questionnaire for auricular reconstruction was used to assess patient satisfaction, and the patient and observer scar assessment scale (POSAS) was used to evaluate scar formation in the surgical area. Data analysis was performed using SPSS 26.0 statistical software. The measurement data were expressed by Mean ± SD, and the counting data were expressed as cases (%). The T-test was used to compare the age difference, length of hospital stay, intraoperative blood loss, scar length, patient satisfaction, and POSAS scores between the two groups. Chi-square test was used to compare the gender composition and incidence of complications between the two groups. P<0.05 was considered statistically significant. Results:A total of 51 patients were included, with 26 in the endoscopic group (14 men and 12 women) and 25 in the open surgery group (12 men and 13 women). The age of the patients in the endoscopic group was (9.8±2.9) years (ranging from 7 to 17 years), while in the open surgery group was (10.3±3.8) years (ranging from 7 to 17 years). The postoperative follow-up period was (15.4±3.4) months (1 to 2 years), and all fascia flaps survived without any severe complications. There were no statistically significant differences between the two groups in terms of age difference, length of hospital stay, intraoperative blood loss, postoperative satisfaction, sex composition ratio, and postoperative complications ( P>0.05). The scar quality in the endoscopy group was superior to that in the open surgery group, and POSAS scores of endoscopic group were lower than those in the open surgery group, and the difference was statistically significant ( P<0.05). Conclusion:Endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia can minimize scarring, improve the postoperative appearance and is not statistically associated with the appearance of reconstructed auricles or complications.
8.An operative position of foot stepping and knee bending in toe replantation for avulsed and fractured great toe
Hongyi YAO ; Yonghui FAN ; Jianfeng LI ; Jianfeng PEI ; Jiebin DUAN ; Kewei ZHANG ; Kangxiong LIANG ; Hongyin LI ; Pengfei WANG ; Yanbing ZHAO ; Guodong TENG
Chinese Journal of Microsurgery 2022;45(1):50-54
Objective:To summarise the advantages and disadvantages of applying the method of retrograde replantation with an operative position of foot stepping and knee bending for replantation of rotational avulsed and fractured great toes.Methods:From January 2016 to June 2021, 11 rotational avulsed and fractured great toes were replanted with the method of retrograde replantation in an operative position of foot stepping and knee bending. Of the 11 patients, 10 were males and 1 was female, aged 18 to 50 years old with an average of 32 years old. Causes of injury: 5 of driving belt, 4 of machinery crush and 2 of car accident. Injury sites: 4 on left great toes and 7 on right great toes. All of the injuries were with fracture and exposure of proximal tendons of flexor and extensor. Seven patients had the follow-up reviews by outpatient clinic visiting, 2 over mobile phone and 2 via WeChat.Results:After surgery, 9 great toes completely survived and 2 great toes had necrosis. The survival rate of toe replantation was 81.8%(9/11). The operation time was 2 to 3 hours, with an average of 2.5 hours. Postoperative X-ray film showed that 8 to 12 weeks after the operation, the fracture and joint fusion were healed at first stage in the survived toes. All patients were entered in follow-up for 3 to 18 months with 10.5 months in average. The survived great toes were plump and the toenail grown well. At the final follow-up, the static TPD at the toes was 8-12 mm, with an average of 10 mm. There was no effect shown on either walking or running.Conclusion:The retrograde replantation method with an operative position of foot stepping and knee bending for great toe replantation has the characteristics of a good field of view and convenient in operation under microscope, a short operation time, and a high survival rate of replantation.
9.Short-term outcome of kidney transplantation from deceased donors with nephrolithiasis
Liang TAN ; Lei SONG ; Yixin XIE ; Longkai PENG ; Gongbin LAN ; Hedong ZHANG ; Hongyi JIANG ; Xubiao XIE
Journal of Central South University(Medical Sciences) 2022;47(9):1217-1226
Objective: Shortage of kidney allografts is a major barrier to end-stage renal disease patients receiving kidney transplantation, and it is necessary to enlarge the donor pool and find better ways of using available allografts. The global incidence of nephrolithiasis is increasing, nephrolithiasis affects approximately 10% of adults worldwide, and it also affects the kidney donors. However, there is little information about the use of cadaveric kidney allografts with nephrolithiasis. This study aims to evaluate the safety and outcome of kidney transplantation with allografts from the deceased donors with nephrolithiasis. Methods: A total of 520 deceased donors who was at least 10 years old, and 945 adult recipients with single kidney transplantation at the Department of Kidney Transplantation, the Second Xiangya Hospital from 2016 to 2020 were included in this study. The donors were divided into 2 groups according to nephrolithiasis diagnoses: The donors with nephrolithiasis (D+) and the donors without nephrolithiasis (D?). The recipients were assigned into 3 groups according to their donors and the allografts they received: The allografts from donors without nephrolithiasis (D?K?), the allografts without nephrolithiasis from donors with nephrolithiasis (D+K?), and the allografts with nephrolithiasis (D+K+). The demographic and clinical data of enrolled subjects were retrospectively analyzed. The allograft discard ratio between different donors were analyzed. The one-year survival of allografts and recipients, as well as the allograft function and the complications of kidney transplantation were compared. Results: Fifty out of 520 donors had nephrolithiasis, and the nephrolithiasis incidence was 9.6%. We recovered 1040 kidneys, and total discard rate was 4.4% (46/1040). The D+ group had a rate of 7% discard. The donors with kidney discard accounted for 12% in the D+ group, and this was higher than that of donors in the D? group (5.1%, P<0.05). The total incidence of delayed graft function (DGF) was 7.5%, and there were no significant differences in the incidence of DGF in recipients among the D?K?, D+K?, and D+K+ group (7.5% vs 6.5% vs 8.2%, P>0.05). During the one-year follow-up, 8 allografts lost function and 19 recipients died with a functional allograft. Recipients in the D?K?, D+K?, and D+K+ groups also had no significant difference between a one-year allograft and patient survival rate (P>0.05). However, recipients in the D+K+ group had a higher level of serum creatinine [(139.2±62.46) μmol/L vs (117.19±51.22) μmol/L, P<0.05] and lower estimated glomerular filtration rate [eGFR; (56.67±23.31) mL/(min·1.73 m?2) vs (66.86±21.90) mL/(min·1.73 m?2), P<0.05] compared with recipients in the D?K? group at 12 months after transplantation. During the first year after transplantation, 4 recipients developed urolithiasis, and recipients who received allografts from the D+ group donors had a higher incidence of urolithiasis than those who received allografts from the D? group donors (2.2% vs 0.2%, P<0.05). There were no significant differences in the incidence of urinary tract infections and ureteral strictures at 1 year between recipients of D+ and D? donors (both P>0.05).Conclusion: The cadaveric kidney allografts with nephrolithiasis could be safely used for transplantation, and the short-term outcome is acceptable. However, nephrolithiasis in donors may increase the rate of kidney discard, disturb the short-term function of allografts, and increase the risk of urolithiasis in recipients. Further research with a long-term study is needed to verify the long-term outcome of kidney transplantation using cadaveric kidney allografts with nephrolithiasis.
10.Analysis of the changes in intestinal microecology in the early stage of sepsis rat based on 16S rDNA sequencing
Hongyi LI ; Ruiqing ZHAI ; Huoyan LIANG ; Yanhui ZHU ; Yan YAN ; Chengyang WANG ; Xianfei DING ; Gaofei SONG ; Tongwen SUN
Chinese Critical Care Medicine 2022;34(1):28-34
Objective:To investigate the changes of intestinal microecology in the early stage of sepsis rat model by 16S rDNA sequencing.Methods:Sixty male Sprague-Dawley (SD) rats were randomly divided into cecal ligation and puncture (CLP) group and sham operation group (Sham group), with 30 rats in each group. In the CLP group, sepsis rat model was reproduced by CLP method; the rats in the Sham group only underwent laparotomy without CLP. At 24 hours after the operation, the intestinal feces and serum samples of 8 rats in each group were collected. The survival rate of the rest rats was observed until the 7th day. The level of serum tumor necrosis factor-α (TNF-α) was detected by enzyme-linked immunosorbent assay (ELISA). Intestinal feces were sequenced by 16S rDNA sequencing technology. The operational taxonomic unit (OTU) data obtained after sequence comparison and clustering was used for α diversity and β diversity analysis, principal coordinate analysis and linear discriminant analysis effect size analysis (LEfSe) to observe the changes of intestinal microecology in early sepsis rats and excavate the marker flora.Results:At 24 hours after the reproduction of the model, the rats in the CLP group showed shortness of breath, scattered hair and other manifestations, and the level of serum TNF-α increased significantly as compared with that in the Sham group (ng/L: 43.95±9.05 vs. 11.08±3.27, P < 0.01). On the 7th day after modeling, the cumulative survival rate of the Sham group was 100%, while that of the CLP group was 31.82%. Diversity analysis showed that there was no significant difference in α diversity parameter between the Sham group and the CLP group (number of species: 520.00±52.15 vs. 492.25±86.61, Chao1 richness estimator: 707.25±65.69 vs. 668.93±96.50, Shannon index: 5.74±0.42 vs. 5.79±0.91, Simpson index: 0.93±0.03 vs. 0.94±0.05, all P > 0.05). However, the β diversity analysis showed that the difference between groups was greater than that within groups whether weighted according to OTU or not (abundance weighted matrix: R = 0.23, P = 0.04; abundance unweighted matrix: R = 0.32, P = 0.01). At the phylum level, the abundance of Proteobacteria and Candidatus_sacchari in the CLP group increased significantly as compared with the Sham group [18.100% (15.271%, 26.665%) vs. 6.974% (2.854%, 9.764%), 0.125% (0.027%, 0.159%)% vs. 0.018% (0.008%, 0.021%), both P < 0.05]. At the genus level, the abundance of opportunistic pathogen including Helicobacter, Ruthenium, Streptococcus, Clostridium ⅩⅧ in the CLP group was significantly higher than that in the Sham group [5.090% (1.812%, 6.598%) vs. 0.083% (0.034%, 0.198%), 0.244% (0.116%, 0.330%) vs. 0.016% (0.008%, 0.029%), 0.006% (0.003%, 0.010%) vs. 0.001% (0%, 0.003%), 0.094% (0.035%, 0.430%) vs. 0.007% (0.003%, 0.030%), all P < 0.05], and the abundance of probiotics such as Alloprevotella and Romboustia was significantly lower than that in the Sham group [7.345% (3.662%, 11.546%) vs. 22.504% (14.403%, 26.928%), 0.113% (0.047%, 0.196%) vs. 1.229% (0.809%, 2.29%), both P < 0.01]. LEfSe analysis showed that the probiotics belonging to Firmicutes were significantly enriched in the Sham group, and Romboustia was the most significantly enriched species. Opportunistic pathogens such as Helicobacter, Streptococcus and Clostridium ⅩⅧ were significantly enriched in the CLP group, Helicobacter_NGSU_ 2015 was the most significantly enriched species. Conclusion:In the early stage of sepsis, the intestinal microbiota structure of rats is significantly changed, which mainly shows that the abundance of Alloprevotella and other probiotics is significantly reduced, while that of Helicobacter and other opportunistic pathogens is significantly increased.

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