1.Construction and validation of a scoring model for pathogen characteristics and short-term prognosis risk prediction of candidemia
Jian-Xin MA ; Xiao-Qiang LIN ; Ming-Chi CAI ; Yu-Zhen XU ; Jun PENG ; Sheng-Qiang LIANG
Medical Journal of Chinese People's Liberation Army 2024;49(3):280-287
Objective To analyze the pathogenic characteristics and drug sensitivity of candidaemia,and construct a short-term mortality risk prediction scoring model.Methods The clinical data of patients with candidaemia admitted to the 909 Hospital of Joint Logistics Support Force from January 2011 to December 2020 were retrospectively analyzed,and the composition of pathogen composition,drug sensitivity test results and incidence of hospitalized patients were analyzed.324 cases of candidaemia were randomly divided into modeling group(190 cases)and validation group(134 cases),and the risk factors were screened by binary logistic regression.According to the odds ratio(OR)score,the 30 day mortality risk prediction scoring model was constructed,and the predictive performance of the model was verified both in modeling and validation groups.Results 356 strains of Candida including 126 strains of C.albicans(35.39%),79 strains of C.tropicalis(22.19%),74 strains of C.parapsilosis(20.79%),48 strains of C.glabrata(13.48%),14 strains of C.guilliermondii(3.93%),8 strains of C.krusei(2.25%),and 7 strains of other Candida(1.97%)were detected in 336 patients with candidemia.The incidence of candidaemia among hospitalized patients increased from 0.20 ‰ in 2011 to 0.48 ‰ in 2020.The resistance rate of candida to amphotericin B was significantly lower than that of fluconazole,voriconazole and itraconazole(P<0.05).Among the 324 cases included in the model,95 patients died in 30 days after diagnosis,and the mortality rate was 29.32%.The proportion of males,fever,and parenteral nutrition in modeling group was significantly higher than that in validation group(P<0.05),while the proportion of chronic lung disease and surgical history within one month were lower than those in validation group(P<0.05).Logistic regression analysis showed that chronic renal failure,mechanical ventilation,severe neutropenia,failure to receive anti-fungal treatment within 72 hours,and APACHE Ⅱ≥20 were risk factors for short-term death of candidaemia,the OR values were 3.179,1.970,2.979,2.080,and 2.399,and the risk scores were 6,4,6,4,and 5,respectively.The area under the curve(AUC)of the risk scoring model for modeling group was 0.792(95%CI 0.721-0.862),and the result of Hosmer-Lemeshow(H-L)test was P=0.305;The AUC of validation group was 0.796(95%CI 0.735-0.898),and the H-L test result was P=0.329.A risk score≤8 indicated a low risk group for short-term mortality,a score of 9-15 indicated a medium risk group,and a score≥16 indicated a high risk group.Conclusions The incidence of candidemia in hospitalized patients is increasing and the mortality is high.The risk prediction score model can effectively predict the short-term prognosis and facilitate the early identification of the prognosis.
2.A preliminary exploration of reduced port laparoscopic proximal gastrectomy with right-sided overlap and single-flap valvuloplasty (ROSF)
Wei PENG ; Qiankun SHAO ; Xinyu LIANG ; Shangcheng YAN ; Qiang CHEN ; Rui REN ; Mengchao SHENG ; Wenting XU ; Yuan TIAN ; Yongyou WU
Chinese Journal of Gastrointestinal Surgery 2024;27(10):1069-1074
Objective:This study aimed to share preliminary experiences of single-incision plus two ports laparoscopic proximal gastrectomy with right-sided overlap and single-flap valvuloplasty (ROSF).Methods:Following the 6th edition of the Japanese Gastric Cancer Treatment Guidelines, proximal gastrectomy with lymphadenectomy was performed. Using a single-port approach, the esophagus was transected at least 2 cm above the tumor's upper margin with linear staplers. The stomach was then extracted through a periumbilical incision, and the proximal stomach was subsequently transected extracorporeally, while ensuring appropriate resection margins on both the greater and lesser curvatures. A single flap was created before returning the remnant stomach to the abdominal cavity and re-establishing pneumoperitoneum. The No.2 clip was used to grasp and elevate the esophageal stump. An incision was made at the right lower edge of the esophageal stump to guarantee that the esophageal lumen was open. The linear stapler was then inserted into the openings of the stomach and esophagus to perform a side overlap anastomosis with a length of 3 cm. Another barbed suture was used to close the common opening of the esophagus and the stomach, and the same barbed suture were used to suture the gastric wall to the lower edge of the muscle flap. The first barbed suture was then used to sequentially suture the proximal brim of the flap to the esophagus and the right brim of the flap to the right brim of the mucosal window. After completion of anastomosis, a drainage tube was inserted through the right upper port. This procedure was employed from November 2023 to March 2024 on five patients diagnosed with adenocarcinoma of the esophagogastric junction and upper stomach. The cohort consisted of three males and two females, with an age range of 62 to 75 years and a body mass index (BMI) of 13.7 to 24.2 kg/m2. All cases were preoperatively staged as T1-2N0M0, confirmed by endoscopic biopsy and enhanced CT scans of the chest, abdomen, and pelvis.Results:All five patients successfully underwent the surgery. The median surgery time was 180-325 minutes, with the intraoperative blood loss of 30-50 ml. The number of lymph nodes harvested ranged from 18 to 27. The time to first flatus, and restore liquid diet and was 2.0-5.0 and 1.0-3.0 days, respectively. The postoperative length of stay was 9.0-11.0 days. The pain scores on the Numeric Rating Scale (NRS). On the first day, the pain scores were 3.0 in two cases, 2.0 in two cases, and 1.0 in one case. On the second day, the pain scores were 2.0 in two cases and 1.0 in three cases. On the third day, the pain scores were 1.0 in four cases and 2.0 in one case. No short-term postoperative complications were observed, and there were no perioperative deaths.Conclusion:Single-incision plus two ports laparoscopic proximal gastrectomy with ROSF is safe and feasible.
3.A preliminary exploration of reduced port laparoscopic proximal gastrectomy with right-sided overlap and single-flap valvuloplasty (ROSF)
Wei PENG ; Qiankun SHAO ; Xinyu LIANG ; Shangcheng YAN ; Qiang CHEN ; Rui REN ; Mengchao SHENG ; Wenting XU ; Yuan TIAN ; Yongyou WU
Chinese Journal of Gastrointestinal Surgery 2024;27(10):1069-1074
Objective:This study aimed to share preliminary experiences of single-incision plus two ports laparoscopic proximal gastrectomy with right-sided overlap and single-flap valvuloplasty (ROSF).Methods:Following the 6th edition of the Japanese Gastric Cancer Treatment Guidelines, proximal gastrectomy with lymphadenectomy was performed. Using a single-port approach, the esophagus was transected at least 2 cm above the tumor's upper margin with linear staplers. The stomach was then extracted through a periumbilical incision, and the proximal stomach was subsequently transected extracorporeally, while ensuring appropriate resection margins on both the greater and lesser curvatures. A single flap was created before returning the remnant stomach to the abdominal cavity and re-establishing pneumoperitoneum. The No.2 clip was used to grasp and elevate the esophageal stump. An incision was made at the right lower edge of the esophageal stump to guarantee that the esophageal lumen was open. The linear stapler was then inserted into the openings of the stomach and esophagus to perform a side overlap anastomosis with a length of 3 cm. Another barbed suture was used to close the common opening of the esophagus and the stomach, and the same barbed suture were used to suture the gastric wall to the lower edge of the muscle flap. The first barbed suture was then used to sequentially suture the proximal brim of the flap to the esophagus and the right brim of the flap to the right brim of the mucosal window. After completion of anastomosis, a drainage tube was inserted through the right upper port. This procedure was employed from November 2023 to March 2024 on five patients diagnosed with adenocarcinoma of the esophagogastric junction and upper stomach. The cohort consisted of three males and two females, with an age range of 62 to 75 years and a body mass index (BMI) of 13.7 to 24.2 kg/m2. All cases were preoperatively staged as T1-2N0M0, confirmed by endoscopic biopsy and enhanced CT scans of the chest, abdomen, and pelvis.Results:All five patients successfully underwent the surgery. The median surgery time was 180-325 minutes, with the intraoperative blood loss of 30-50 ml. The number of lymph nodes harvested ranged from 18 to 27. The time to first flatus, and restore liquid diet and was 2.0-5.0 and 1.0-3.0 days, respectively. The postoperative length of stay was 9.0-11.0 days. The pain scores on the Numeric Rating Scale (NRS). On the first day, the pain scores were 3.0 in two cases, 2.0 in two cases, and 1.0 in one case. On the second day, the pain scores were 2.0 in two cases and 1.0 in three cases. On the third day, the pain scores were 1.0 in four cases and 2.0 in one case. No short-term postoperative complications were observed, and there were no perioperative deaths.Conclusion:Single-incision plus two ports laparoscopic proximal gastrectomy with ROSF is safe and feasible.
4.The correlation between colorectal cancer tissues JAK2 gene mutations and TCF3 protein expression with clinical pathological features and prognosis
Ke-Jin YAN ; Hong-Liang ZHU ; Xian-Qiang XIE ; Dong-Sheng LI ; Zeng-Lu ZHENG ; Ji-Yang LI
Modern Interventional Diagnosis and Treatment in Gastroenterology 2024;29(6):675-680,690
Objective To explore the correlation between colorectal cancer tissue Janus kinase 2(JAK2)gene mutations and T cytokine 3(TCF3)protein expression with clinical pathological characteristics and prognosis,and to provide laboratory reference indicators for early evaluation of the illness severity and prognosis of colorectal cancer.Methods A retrospective analysis was conducted on the data of 50 colorectal cancer patients who were admitted from January 2016 to April 2021 and retained colon cancer and adjacent tissue wax blocks.Basic information,clinical and pathological features such as TNM staging,lymph node metastasis,and 3-year survival prognosis of the patients were collected.The wax blocks of colon cancer and adjacent tissues of patients were detected,in which Taqman fluorescence probe method was applied to detect the distribution of JAK2 gene at the rs2230724 locus AA,AG,and GG genotypes in colon cancer tissues,and immunohistochemistry method was applied to detect the positive expression rate of TCF3 protein in colon cancer and adjacent tissues.The basic information,JAK2 rs2230724 gene mutation,and TCF3 protein expression of patients with different clinical and pathological characteristics were compared,and the influencing factors of clinical and pathological characteristics of colon cancer was analyzed by logistic regression model.Kaplan Meier survival curve was applied to compare the survival prognosis of patients with JAK2 gene mutations and TCF3 protein expression in colorectal cancer tissue,and Cox regression model was applied to analyze the risk factors affecting the prognosis of colorectal cancer patients.Results The positive expression rate of TCF3 protein in colon cancer tissues was higher than that in adjacent tissues(P<0.05).The age,BMI,proportion of GG genotype at rs2230724 locus of JAK2 gene and positive expression rate of TCF3 protein in TNM stage Ⅲ colon cancer patients were higher than those in TNM stage Ⅰ-Ⅱ patients(P<0.05);The age,BMI,smoking rate,proportion of GG type at the rs2230724 locus of JAK2 gene in colon cancer tissue,and positive expression rate of TCF3 protein in patients with lymph node metastasis were higher than those without lymph node metastasis(P<0.05);The results of the logistic regression model analysis showed that the influencing factors of clinical pathological features such as TNM staging and lymph node metastasis in colon cancer were age,mutation of JAK2 gene rs2230724 site in colon cancer tissue,and positive expression rate of TCF3 protein(P<0.05).The Kaplan Meier survival curve analysis showed that patients with JAK2 gene rs2230724 GG genotype and TCF3 protein positivity in colon cancer tissue had higher cumulative all-cause mortality rates(P<0.05).The results of univariate and multivariate Cox regression model analysis showed that independent risk factors affecting the prognosis of colorectal cancer patients include JAK2 gene rs2230724 site GG type,TCF3 protein positive expression,TNM stage Ⅲ,lymph node metastasis,and age.Conclusion The proportion of JAK2 gene rs2230724 GG type and TCF3 protein expression in colorectal cancer tissues are related to their clinical pathological characteristics and prognosis,and can be used as reference indicators for evaluating clinical pathological characteristics and predicting prognosis of colorectal cancer.
5.Single posterior osteotomy for the treatment of rigid cervical spine deformities
Xiaoyu CAI ; Tao XU ; Maimaiti MAIERDAN· ; Rui CAO ; Chuanhui XUN ; Weidong LIANG ; Jian ZHANG ; Qiang DENG ; Maimaiti PULATI· ; Jun SHENG ; Ting WANG ; Weibin SHENG
Chinese Journal of Orthopaedics 2024;44(19):1265-1272
Objective:To evaluate the safety and effectiveness of single posterior osteotomy in the correction of rigid cervical spine deformities (CSD) and to explore the indications and key surgical techniques involved.Methods:A retrospective analysis was conducted on the clinical data of 9 patients with rigid CSD who underwent single posterior osteotomy correction between June 2012 and June 2023 in the Department of Spine Surgery at the First Affiliated Hospital of Xinjiang Medical University. The cohort comprised 4 males and 5 females, with a mean age of 19.8±27.2 years (range, 7-48 years). Among these, 5 cases were congenital CSD, 3 were post-tuberculosis deformities, and 1 was iatrogenic. Various coronal and sagittal alignment parameters were measured, including C 1, 2 angle, cervical lordosis (CL), structural scoliosis angle (SSA), structural kyphosis angle (SKA), head tilt (HT), C 2-C 7 sagittal vertical axis (CSVA), sagittal vertical axis (SVA), coronal balance distance (CBD), T 1 slope (T 1S), and the difference between T 1 tilt and cervical lordosis (T 1S-CL). Clinical outcomes were assessed using the neck disability index (NDI), visual analogue scale (VAS), and Scoliosis Research Society-22 questionnaire (SRS-22). Results:The average operation time was 273.9±76.1 min, with an average blood loss of 472.2±128.8 ml. All 9 patients were followed up for an average of 45.2±41.8 months (range, 12-116 months). A total of 7 patients underwent single-segment osteotomies (C 3, C 6 and C 7: 1 case each; C 5: 4 cases), and 2 patients underwent double-segment osteotomies (C 2 and C 7, C 3 and C 4). Four cases involved pedicle subtraction osteotomy (PSO), while 7 cases required vertebral column resection. The upper instrumented vertebra (UIV) was located at the occiput in 1 case and in the cervical spine in 8 cases. The lower instrumented vertebra (LIV) was located in the upper thoracic spine in 6 cases and in the cervical spine in 3 cases, with 2 of the latter cases having both UIV and LIV in the cervical spine. The average number of fused segments was 7.6±4.4 segments (range, 2-12 segments). All patients achieved successful bone fusion within an average of 8.8±3.2 months (range, 6-12 months). Preoperatively, the mean values for CL, SSA, SKA, HT, and CBD were 19.8° (17.2°, 30.5°), 27.4°(23.3°, 30.4°), 28.4°(25.6°, 30.1°), 9.0°(6.2°, 12.3°), and 18.5(12.3, 23.6) mm, respectively. Postoperative improvements were noted with values of -11.1°(-8.8°, -14.4°), 1.3°(0.8°, 1.6°), -11.1°(-8.6°, -14.5°), 1.6°(0.5°, 2.2°), and 9.4 (4.8-13.5) mm, respectively. At the final follow-up, these parameters were maintained, with values of -11.0°(-8.8°, -14.3°), 1.2°(0.8°, 1.5°), -11.0° (-8.6°, -14.3°), 1.5°(0.5°, 2.2°), and 9.4(4.8, 13.4) mm, respectively. Statistically significant improvements were observed between preoperative and postoperative measurements ( P<0.05), except for C 1, 2 angle, CSVA, SVA, T 1S, and T 1S-CL ( P>0.05). NDI and SRS-22 scores showed significant improvements postoperatively ( P<0.05), while VAS scores did not show a significant change ( P>0.05). Postoperative complications included transient nerve injury in two patients, one case of right central retinal artery occlusion, and one case of vertebral artery injury. Conclusion:This study confirms the safety and efficacy of single posterior osteotomy for treating rigid CSD of various etiologies. Standard PSO or modified techniques are effective for correcting cervical kyphosis, while hemivertebra resection and concave-side distraction are recommended for congenital scoliosis or kyphoscoliosis.
6.Protective Effect of Shenfu Injection on Vascular Endothelial Damage in a Porcine Model of Hemorrhagic Shock.
Ming-Qing ZHANG ; Qiang ZHANG ; Wei YUAN ; Jun-Yuan WU ; Yong LIANG ; Hong-Jie QIN ; Chun-Sheng LI
Chinese journal of integrative medicine 2022;28(9):794-801
OBJECTIVE:
To investigate the effects of Shenfu Injection (, SFI) on endothelial damage in a porcine model of hemorrhagic shock (HS).
METHODS:
After being bled to a mean arterial pressure of 40±3 mm Hg and held for 60 min, 32 pigs were treated with a venous injection of either shed blood (transfusion group), shed blood and saline (saline group), shed blood and SFI (SFI group) or without resuscitation (sham group). Venous blood samples were collected and analyzed at baseline and 0, 1, 2, 4, and 6 h after HS. Tumor necrosis factor-α (TNF-α), serum interleuking (IL)-6, and IL-10 levels were measured by enzyme-linked immunosorbent assay (ELISA); expressions of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule 1 (ICAM -1), von Willebrand factor (vWF), plasminogen activator inhibitor-1 (PAI-1) and Bcl-2, Bax, and caspase-3 proteins were determined by Western blot.
RESULTS:
The serum level of TNF-α in the SFI group was significantly lower than in the other groups at 0, 1, and 2 h after HS, while the level of IL-6 was lower at 4 and 6 h compared with the saline group (P<0.01 or P<0.05). The concentration of serum IL-10 was significantly higher in the SFI group than in the other groups at 0, 1, 4, and 6 h after HS (P<0.01). Western blot and immunohistochemistry of vascular tissue showed that the expression of caspase-3 was downregulated, and that of Bcl-2 and Bax was upregulated in the SFI group compared to other groups (P<0.05).
CONCLUSION
SFI attenuated endothelial injury in the porcine model of HS by inhibiting cell apoptosis, suppressing the formation of proinflammatory cytokines, and reducing endothelial activation.
Animals
;
Caspase 3/metabolism*
;
Drugs, Chinese Herbal
;
Interleukin-10
;
Proto-Oncogene Proteins c-bcl-2/metabolism*
;
Shock, Hemorrhagic/drug therapy*
;
Swine
;
Tumor Necrosis Factor-alpha/metabolism*
;
bcl-2-Associated X Protein/metabolism*
7.Danggui Buxuetang Alleviates Oxidative Stress and Inflammation in Diabetic Kidney Disease Rats by Improving Mitochondrial Dysfunction of Podocytes
He-chao JIN ; Jia-wei QIANG ; Guan-wen ZHANG ; Sheng-ran LIANG ; Deng-zhou GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):31-40
ObjectiveTo investigate the intervention effect of Danggui Buxuetang on oxidative stress and inflammatory response in diabetic kidney disease (DKD) rats from its improvement of podocyte mitochondrial dysfunction. MethodSD rats were randomly divided into the control group and modeling group, and the ones in the latter group rats were fed a high-glucose and high-fat diet and then intraperitoneally injected with a small dose of streptozotocin (STZ) for inducing type 2 diabetes. The successfully modeled rats were randomized into the model group, high- and low-dose (1.44 and 0.72 g·kg-1) Danggui Buxuetang groups, and irbesartan (0.017 g·kg-1)group and gavaged with the corresponding drugs, while those in the normal and model groups with an equal volume of normal saline. After 20 weeks of drug intervention, the urinary microalbumin-to-urine creatinine ratio (UACR) and serum malondialdehyde (MDA) content and manganese superoxide dismutase (MnSOD) activity in each group were measured. The pathological changes in renal tissue were observed by Masson trichrome staining, and periodic acid-silver metheramine (PASM) staining, followed by the observation of ultrastructural changes in podocytes under the transmission electron microscope (TEM). The expression level of reactive oxygen species (ROS) in rat kidney tissue was detected using a fluorescent probe dihydroethidium (DHE). The protein expression levels of peroxisome proliferator-activated receptor γ -coactivator -1α (PGC-1α), nucleotide-binding domain like receptor protein 3 (NLRP3), and Wilms tumor protein-1 (WT-1) were measured by immunohistochemistry (IHC), and the expression levels of NLRP3, interleukin-1β (IL-1β),and WT-1 in podocytes by immunofluorescence (IF) assay. The mRNA expression levels of PGC-1α and NLRP3 in the renal tissues were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expression levels of PGC-1α, MnSOD, NLRP3, and IL-1β were assayed by Western blot. ResultCompared with the normal group, the model group exhibited elevated UACR and MDA content, weakened MnSOD activity (P<0.01), glomerular hypertrophy, thickened basement membrane, mesangial hyperplasia, increased extracellular matrix, K-W nodules, podocyte mitochondrial swelling, disordered mitochondrial cristae, foot process fusion or loss, vacuolization, increased ROS (P<0.01), enhanced NLRP3 and IL-1β but diminished WT-1 expression in podocytes, down-regulated PGC-1α mRNA expression (P<0.01) and PGC-1α and MnSOD protein expression (P<0.01), and up-regulated NLRP3 mRNA expression and NLRP3 and IL-1β protein expression (P<0.01). Compared with the model group, Danggui Buxuetang high-dose group significantly decreased UACR and MDA, enhanced MnSOD activity (P<0.05, P<0.01), improved renal histopathology and podocyte mitochondrial ultrastructure, decreased ROS (P<0.05, P<0.01) and NLRP3 and IL-1β expression in podocytes, enhanced WT-1 expression in podocytes, up-regulated the mRNA and protein levels of PGC-1α and MnSOD, and down-regulated the mRNA and protein levels of NLRP3 and IL-1β (P<0.05, P<0.01). ConclusionDanggui Buxuetang alleviates oxidative stress, reduces inflammatory response, protects kidney, and delays the progression of DKD possibly by improving the mitochondrial dysfunction in podocytes of DKD rats.
8.Effect of Danggui Buxuetang on Podocyte Injury in Diabetic Kidney Disease Rats: An Exploration Based on RIPK1/RIPK3/MLKL Signaling Pathway
He-chao JIN ; Guan-wen ZHANG ; Sheng-ran LIANG ; Jia-wei QIANG ; Deng-zhou GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):41-48
ObjectiveTo observe the effect of Danggui Buxuetang on the podocyte injury and receptor-interacting protein kinase 1/receptor-interacting protein kinase3/mixed lineage kinase domain-like protein (RIPK1/RIPK3/MLKL) signaling pathway in diabetic kidney disease (DKD) ratsand to explore its possible mechanism against DKD. MethodEight of the 50 SD rats were randomly classified intoa normal group, and the remaining were fed a high-glucose and high-fat diet for six weeks and then intraperitoneally injected with 0.035 g·kg-1streptozotocin (STZ) for inducing type 2 diabetes. After successful modeling,they were randomized into the model group,high- and low-dose (1.44,0.72 g·kg-1) Danggui Buxuetang groups, and irbesartan (0.017 g·kg-1)group. After 20 weeks of drug intervention, the fasting blood glucose (FBG), kidney index (KI),and urinary microalbumin-to-urine creatinine ratio (UACR)were detected in each group. The pathological changes in renal tissue were observed by hematoxylin-eosin (HE) staining, followed by the observation of ultrastructural changes in podocytes under the transmission electron microscope. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in renal tissue of rats were determined by enzyme-linked immunosorbent assay (ELISA), and the protein expression levels of RIPK1, RIPK3, and MLKL in rat kidney tissue by immunohistochemistry. The apoptosis rate of podocytes was detected by in situ nick end-labeling (TUNEL) assay. The mRNA expression levels of RIPK1, RIPK3, and MLKL in kidney tissue of rats were measured by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expression levels of RIPK, RIPK3, and MLKL and podocyte marker Wilms tumor protein-1 (WT-1) in rat kidney tissue were assayed by Western blot. ResultCompared with the normal group, the model group exhibited elevated FBG, UACR, and KI (P<0.01), glomerular hypertrophy, thickened basement membrane, increased extracellular matrix, mesangial hyperplasia, foot process fusion or loss, enhanced apoptosis in renal tissue, up-regulated TNF-α and IL-6 levels (P<0.01) and RIPK1/RIPK3/MLKL mRNA and protein expression (P<0.01), and down-regulated WT-1 protein expression. Compared with the model group, Danggui Buxuetang high-dose group significantly reduced the levels of FBG, UACR, and KI, improved renal histopathology, podocyte loss, and apoptosis in renal tissue, down-regulated TNF-α and IL-6 levels and RIPK1/RIPK3/MLKL mRNA and protein expression (P<0.05, P<0.01), and up-regulated WT-1 protein expression. ConclusionDanggui Buxuetang alleviates podocyte injury and delays the development of DKD possibly by regulating the RIPK1/RIPK3/MLKL signaling pathway.
9.Effect of Danggui Buxuetang on Podocyte Pyroptosis in Diabetic Kidney Disease Rats: An Exploration Based on TXNIP/NLRP3/GSDMD Signaling Pathway
He-chao JIN ; Sheng-ran LIANG ; Guan-wen ZHANG ; Jia-wei QIANG ; Deng-zhou GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(3):49-57
ObjectiveTo observe the effect of Danggui Buxuetang on podocyte pyroptosis in diabetic kidney disease (DKD) rats and to explore the possible mechanism of its prevention and treatment of DKD and podocyte pyroptosis. MethodEight of the 50 male SD rats were randomly classified into a normal group, and the remaining 42 were fed a high-glucose and high-fat diet for six weeks and then intraperitoneally injected with 35 mg·kg-1 streptozotocin (STZ) for inducing type 2 diabetes. After successful modeling, they were randomized into the model group, low- (0.72 g·kg-1) and high-dose (1.44 g·kg-1) Danggui Buxuetang group, and irbesartan (0.017 g·kg-1) group and gavaged with the corresponding drugs, while those in the normal group and model group with an equal volume of normal saline, once per day, for 20 weeks. During the medication, the fasting blood glucose (FBG) and 24 h urine protein (24 h-UTP) were measured regularly. After administration, the pathological changes in renal tissues were observed by periodic acid-silver metheramine (PASM) staining, followed by the observation of ultrastructural changes in podocytes under the transmission electron microscope (TEM). Serum levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) were determined by enzyme-linked immunosorbent assay (ELISA). The DNA damage in renal tissue cells of rats was detected by in situ nick end-labeling (TUNEL) assay. The protein expression levels of thioredoxin interacting protein (TXNIP), cysteine-dependent aspartate-directed protease-1 (Caspase-1), and gasdermin D (GSDMD) in renal tissues of rats were detected by immunohistochemistry (IHC), the expression levels of nucleotide binding domain like receptor protein 3 (NLRP3) and Wilms tumor protein-1 (WT-1) in podocytes by immunofluorescent (IF) staining, and the expression levels of TXNIP/NLRP3/Caspase-1/GSDMD pathway proteins and Synaptopodin in renal podocytes by Western blot. ResultCompared with the normal group, the model group exhibited increased FBG and 24 h UTP, glomerular hypertrophy, mesangial hyperplasia, increased extracellular matrix, thickened basement membrane, K-W nodules, vacuolar degeneration in renal tubular epithelial cells, foot process fusion or loss, elevated serum IL-1β and IL-18 levels and TUNEL-positive cells in renal tissue, enhanced NLRP3 but diminished WT-1 expression in podocytes, down-regulated Synaptopodin protein expression, and up-regulated TXNIP/NLRP3/Caspase-1/GSDMD protein expression (P<0.01). Compared with the model group, Danggui Buxuetang high-dose group remarkably lowered FBG, 24-h UTP, and TUNEL-positive cells in renal tissue, improved renal histopathology and podocyte injury and loss, down-regulated NLRP3 expression in podocytes and TXNIP/NLRP3/Caspase-1/GSDMD protein expression levels, and up-regulated WT-1 expression in podocytes and Synaptopodin protein expression (P<0.05, P<0.01). ConclusionDanggui Buxuetang inhibits podocyte pyroptosis to reduce proteinuria and delays the development of DKD possibly by regulating the TXNIP/NLRP3/GSDMD signaling pathway.
10.Establishment of an animal model of Sparganum mansoni infection and study on therapeutic methods II Establishment of a mouse model of sparganosis mansoni via oral administration of procercoids
Xian-shi TANG ; Yong-liang XU ; Qiang WANG ; Ying ZHANG ; De-sheng TONG
Chinese Journal of Schistosomiasis Control 2022;34(4):400-403
Objective To establish an animal model of sparganosis mansoni through oral administration of Cyclops infected with procercoids. Methods Domestic cats were infected with Sparganum mansoni under laboratory conditions, and fresh cat stool samples were collected, washed in dechlorinated water, and filtered. Spirometra mansoni eggs were collected and prepared into suspensions. Twenty C57BL/6j mice were randomly divided into the experimental group (n = 15) and the control group (n = 5). Wild Cyclops were infected with Spirometra mansoni coracidia to allow 3 to 5 procercoids in each Cyclop. Then, each mouse in the experimental group was given 15 Cyclops infected with procercoids by gavage, while mice in the control group were orally administered with the same volume of dechlorinated water. All mice were sacrificed after 5 months, and dissected, and suspicious Sparganum mansoni worms were collected. The serum specific IgG antibody against Sparganum mansoni was measured in mice using enzyme-linked immunosorbent assay (ELISA). Genomic DNA was isolated from suspicious Sparganum mansoni worms, and the specific Sparganum mansoni cytochrome oxidase I (COI) gene was amplified using PCR assay. Results Among the 15 mice in the experimental group, six were positive for the serum specific IgG antibody against Sparganum mansoni, and milky white worms were found and collected from the subcutaneous regions of 4 out of 6 mice. Only one worm was detected in each mouse, and the worm morphology was similar to Sparganum mansoni. Capillary electrophoresis of the PCR amplification products of COI gene presented a specific band with 151 bp in size, and sequencing analysis revealed 100% homology with Sparganum mansoni. Conclusions A mouse model of sparganosis mansoni is successfully created through oral administration of Cyclops infected with Spirometra mansoni procercoids.

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