1.Establishment of Allogeneic Kidney Transplantation Technical System in Banna Miniature Pig Inbred Strain
Ying ZHANG ; Ziwei PENG ; Chang YANG ; Jing WANG ; Hongfang ZHAO ; Gen CHEN ; Hongye ZHAO ; Hongjiang WEI
Laboratory Animal and Comparative Medicine 2025;45(5):623-633
Objective To establish a technical system for allogeneic kidney transplantation surgery in pigs using the Banna miniature pig inbred strain,and to evaluate it through routine blood tests,liver and kidney function tests,thus providing reference data for the preparation of allogeneic kidney transplantation models.Methods A total of 4 cases of allogeneic kidney transplantation surgeries were performed,including 1 case of single kidney transplantation in a healthy pig,2 cases of kidney transplantation after unilateral nephrectomy,and 1 case of kidney transplantation after bilateral nephrectomy.Before kidney transplantation,cross-matching and complement-dependent cytotoxicity(CDC)tests were used for matching between donor and recipient pigs.After kidney transplantation,peripheral blood samples were regularly collected from pigs for routine blood tests,liver function tests,and kidney function tests,and color Doppler ultrasound technology was used to detect blood supply to the transplanted kidneys.After reaching the experimental endpoint,both kidneys of pig DR1 and the left kidney of pig DR3 were collected and hematoxylin-eosin(HE)staining was performed to evaluate pathological changes in the transplanted kidneys.Results Recipient pigs DR1 and DR3 died at 17 days and 30 days after surgery respectively,while recipient pigs R and DR2 remained in good condition during the 30-day observation period.The results of liver and kidney function test showed that in pig DR1,alanine aminotransferase(ALT)levels increased on postoperative day 1(>1 000 U/L),peaked on postoperative day 7(1 300 U/L),and aspartate aminotransferase(AST)levels peaked on postoperative day 1(>3 000 U/L).On postoperative day 17,ALT and AST levels remained high(ALT,500 U/L;AST,700 U/L).In pigs R,DR2,and DR3,ALT and AST levels returned to normal around day 17.Serum creatinine(Crea)levels in pig R remained stable without postoperative increase.Crea levels in pigs DR1 and DR2 showed transient elevation on postoperative day 1,then gradually returned to normal(<100 μmol/L).Crea levels in pig DR3 remained below 500 μmol/L from postoperative days 2-10,but increased between days 11-28,reaching up to 1 500 μmol/L,indicating gradual loss of kidney function.Ultrasound results showed that the preoperative resistive index(RI)of recipient pig R was 0.91.On postoperative day 24,renal cortex and medulla showed abundant blood flow signals with RI value of 0.88,which was close to the pre-transplantation RI value.For pig DR2,the RI value on postoperative day 17 was 0.89,with poor renal cortex blood flow and relatively good renal medulla blood flow.In pig DR1 on postoperative day 17,no blood flow signals were detected in the transplanted kidney.HE staining results showed that the non-transplanted healthy right kidney of pig DR1 had normal structure,while the transplanted left kidney showed blurred glomerular structure and nuclear dissolution,indicating that the left kidney had lost function before removal.In the transplanted left kidney of pig DR3,large numbers of red blood cells and lymphocyte infiltration were observed in glomeruli and renal tubules,indicating possible coagulation dysfunction and rejection reactions after kidney transplantation.Conclusion Banna miniature pig inbred strain is used as experimental animals to perform four cases of allogeneic pig-to-pig kidney transplantation.The physiological parameters of the recipient pig and the function of the transplanted kidney are monitored after surgery using routine blood tests,liver and kidney function tests,color Doppler ultrasound,and pathological examinations.The allogeneic pig-to-pig kidney transplantation technical system established in the study can provide a foundation for clinicians to conduct kidney transplantation surgeries.
2.Establishment of Allogeneic Kidney Transplantation Technical System in Banna Miniature Pig Inbred Strain
Ying ZHANG ; Ziwei PENG ; Chang YANG ; Jing WANG ; Hongfang ZHAO ; Gen CHEN ; Hongye ZHAO ; Hongjiang WEI
Laboratory Animal and Comparative Medicine 2025;45(5):623-633
Objective To establish a technical system for allogeneic kidney transplantation surgery in pigs using the Banna miniature pig inbred strain,and to evaluate it through routine blood tests,liver and kidney function tests,thus providing reference data for the preparation of allogeneic kidney transplantation models.Methods A total of 4 cases of allogeneic kidney transplantation surgeries were performed,including 1 case of single kidney transplantation in a healthy pig,2 cases of kidney transplantation after unilateral nephrectomy,and 1 case of kidney transplantation after bilateral nephrectomy.Before kidney transplantation,cross-matching and complement-dependent cytotoxicity(CDC)tests were used for matching between donor and recipient pigs.After kidney transplantation,peripheral blood samples were regularly collected from pigs for routine blood tests,liver function tests,and kidney function tests,and color Doppler ultrasound technology was used to detect blood supply to the transplanted kidneys.After reaching the experimental endpoint,both kidneys of pig DR1 and the left kidney of pig DR3 were collected and hematoxylin-eosin(HE)staining was performed to evaluate pathological changes in the transplanted kidneys.Results Recipient pigs DR1 and DR3 died at 17 days and 30 days after surgery respectively,while recipient pigs R and DR2 remained in good condition during the 30-day observation period.The results of liver and kidney function test showed that in pig DR1,alanine aminotransferase(ALT)levels increased on postoperative day 1(>1 000 U/L),peaked on postoperative day 7(1 300 U/L),and aspartate aminotransferase(AST)levels peaked on postoperative day 1(>3 000 U/L).On postoperative day 17,ALT and AST levels remained high(ALT,500 U/L;AST,700 U/L).In pigs R,DR2,and DR3,ALT and AST levels returned to normal around day 17.Serum creatinine(Crea)levels in pig R remained stable without postoperative increase.Crea levels in pigs DR1 and DR2 showed transient elevation on postoperative day 1,then gradually returned to normal(<100 μmol/L).Crea levels in pig DR3 remained below 500 μmol/L from postoperative days 2-10,but increased between days 11-28,reaching up to 1 500 μmol/L,indicating gradual loss of kidney function.Ultrasound results showed that the preoperative resistive index(RI)of recipient pig R was 0.91.On postoperative day 24,renal cortex and medulla showed abundant blood flow signals with RI value of 0.88,which was close to the pre-transplantation RI value.For pig DR2,the RI value on postoperative day 17 was 0.89,with poor renal cortex blood flow and relatively good renal medulla blood flow.In pig DR1 on postoperative day 17,no blood flow signals were detected in the transplanted kidney.HE staining results showed that the non-transplanted healthy right kidney of pig DR1 had normal structure,while the transplanted left kidney showed blurred glomerular structure and nuclear dissolution,indicating that the left kidney had lost function before removal.In the transplanted left kidney of pig DR3,large numbers of red blood cells and lymphocyte infiltration were observed in glomeruli and renal tubules,indicating possible coagulation dysfunction and rejection reactions after kidney transplantation.Conclusion Banna miniature pig inbred strain is used as experimental animals to perform four cases of allogeneic pig-to-pig kidney transplantation.The physiological parameters of the recipient pig and the function of the transplanted kidney are monitored after surgery using routine blood tests,liver and kidney function tests,color Doppler ultrasound,and pathological examinations.The allogeneic pig-to-pig kidney transplantation technical system established in the study can provide a foundation for clinicians to conduct kidney transplantation surgeries.
3.Implication of Qidi Tangshen Prescription (QDTS) on Podocyte Pyroptosis in Diabetes Nephropathy by Regulating MAPK14/RELA/Caspase-8 Signaling Pathway
Fei GAO ; Borui YU ; Huidi XIE ; Ying ZHOU ; Yang SHI ; Xianhui ZHANG ; Hongfang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):67-75
ObjectiveTo explore the molecular mechanism of Qidi Tangshen prescription (QDTS) in regulating podocyte pyroptosis in diabetes nephropathy (DN). MethodThrough in vivo experiment, db/db mice were divided into the model group, QDTS group (3.34 g·kg-1), valsartan capsule group (10.29 mg·kg-1), with db/m mice serving as the normal control. Each group consisted of 8 mice, and they underwent continuous intervention for 8 weeks. After the last administration, mice were euthanized, and kidney pathological changes were observed. Additionally, the expression levels of pyroptosis-related indicators, including NOD-like receptor protein 3 (NLRP3), Gasdermin D protein (GSDMD), and interleukin-1β (IL-1β) protein, were examined. Through in vitro experiment, mouse podocytes were divided into the normal glucose group (5.5 mmol·L-1 glucose), high glucose group (35 mmol·L-1 glucose), DMSO group (35 mmol·L-1 glucose+200 mg·L-1 DMSO), and QDTS group (35 mmol·L-1 glucose+200 mg·L-1 QDTS freeze-dried powder). After 48 hours of intervention, the expression levels of NLRP3, GSDMD, and IL-1β proteins were measured in podocytes. A drug-ingredient-target-disease interaction network for QDTS in the treatment of DN was constructed by network pharmacology methods. The key signaling pathways regulating podocyte pyroptosis were analyzed, and validation was conducted through in vivo and in vitro experiments. ResultCompared with normal group, glomerular hyperplasia and glomerular basement membrane thickening were observed in model group, and some segments were accompanied by obvious podocellular process fusion. The protein expressions of NLRP3, GSDMD and IL-1β in mouse kidney were increased, the protein expressions of mitogen-activated protein kinase 14 (MAPK14), V-Rel reticuloendotheliosis virus oncogene homology A (RELA) and Caspase-8 in mouse kidney were increased (P<0.05). Compared with model group, kidney pathological injury of mice in QDTS group was significantly reduced, and the expressions of NLRP3, GSDMD and IL-1β in kidney of mice in QDTS group and valsartan group were decreased (P<0.05). The protein expressions of MAPK14, RELA and Caspase-8 in kidney of mice in QDTS group and valsartan group were decreased (P<0.05). Network pharmacology results showed that there were 16 targets for QDTS to regulate DN cell pyrodeath, among which MAPK14, RELA and Caspase-8 were the key targets. Compared with normal glucose group, the protein expressions of NLRP3, GSDMD and IL-1β in high glucose group were increased (P<0.05), and the protein expressions of MAPK14, RELA and Caspase-8 in mouse podocytes were increased (P<0.05). Compared with high glucose group, the expressions of NLRP3, GSDMD and IL-1β in podocytes of mice in QDTS group were decreased (P<0.05), and the expressions of MAPK14, RELA and Caspase-8 in podocytes of mice in QDTS group were decreased (P<0.05). ConclusionQDTS reduces damage to DN podocytes, which is associated with its regulation of the MAPK14/RELA/Caspase-8 signaling pathway and inhibition of podocyte pyroptosis.
4.Qidi Tangshen Prescription (QDTS) Regulate Akt1/HIF-1α/Bcl-xl Signaling Pathway to Improve Podocyte Autophagy in Diabetic Nephropathy
Fei GAO ; Huidi XIE ; Borui YU ; Ying ZHOU ; Yang SHI ; Xianhui ZHANG ; Hongfang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):90-97
ObjectiveTo explore the mechanism of Qidi Tangshen prescription (QDTS) in alleviating podocyte injury and reducing urinary protein in diabetic nephropathy (DN). MethodUsing network pharmacology methods, we collected the chemical components and targets of QDTS, as well as the targets related to DN. Subsequently, we constructed a "drug-ingredient-target-disease" network for QDTS in the treatment of DN to systematically elucidate the mechanism. The db/db mice were assigned into the model, QDTS (3.34 g·kg-1), and losartan capsules (10.29 mg·kg-1) groups, and db/m mice served as the normal group. Each group consisted of 8 mice, and they underwent continuous intervention for 8 weeks. After the last administration, mice were euthanized, and the urinary albumin excretion rate (UAER) and renal pathological changes were measured and observed. The expression levels of protein kinase B1 (Akt1), hypoxia-inducible factor-1 alpha (HIF-1α), phosphorylated B-cell lymphoma-extra-large (p-Bcl-xl), as well as autophagy-related indicators microtubule-associated protein 1 light chain 3 (LC3), ubiquitin-binding protein p62 (p62), and autophagy-related gene 6 homolog (Beclin1), were determined. Furthermore, mouse podocytes were divided into the normal glucose (5.5 mmol·L-1), high glucose (35 mmol·L-1), DMSO (35 mmol·L-1 glucose+200 mg·L-1 DMSO), and QDTS (35 mmol·L-1 glucose+200 mg·L-1 QDTS freeze-dried powder) groups. After 48 h of intervention, the protein levels of Akt1, HIF-1α, p-Bcl-xl, LC3, p62, and Beclin1 in podocytes were measured. ResultQDTS had 34 active components acting on 143 targets in the treatment of DN, and 55 targets were related to autophagy, in which Akt1, HIF-1α, and Bcl-xl were the key targets. Compared with the normal group, mice in the model group exhibited significantly increased UAER, glomerular hypertrophy, deposition of blue collagen fibers, thickening of the glomerular basement membrane, and noticeable fusion of podocyte foot processes in some segments. Furthermore, the modeling up-regulated the protein levels of p-Akt1, HIF-1α, and p62 and down-regulating the protein levels of p-Bcl-xl, LC3, and Beclin1 in the renal tissue (P<0.05). Compared with the model group, QDTS and losartan decreased UAER (P<0.05) and alleviated the pathological damage in the renal tissue. Moreover, QDTS and losartan down-regulated the protein levels of p-Akt1, HIF-1α, and p62 and up-regulated the protein levels of p-Bcl-xl, LC3, and Beclin1 in the renal tissue (P<0.05). In comparison to the normal glucose group, the high glucose group displayed up-regulated protein levels of p-Akt1, HIF-1α, and p62 and down-regulated protein levels of p-Bcl-xl, LC3, and Beclin1 in podocytes (P<0.05). Compared with the high glucose group, QDTS down-regulated the protein levels of p-Akt1, HIF-1α, and p62 and up-regulated the protein levels of p-Bcl-xl, LC3, and Beclin1 in podocytes (P<0.05). ConclusionQDTS alleviates podocyte damage and reduced urinary protein in DN by regulating the Akt1/HIF-1α/Bcl-xl signaling pathway, thereby enhancing podocyte autophagy.
5.Abdominal pregnancy in the sac of lesser omentum after in vitro fertilization and embryo transfer: a case report
Jinghui JIA ; Bing CHEN ; Mengnan YU ; Hongfang WANG ; Ying TONG ; Ying SHI
Chinese Journal of Reproduction and Contraception 2023;43(4):404-406
Objective:To summarize the clinical experience in the diagnosis and treatment of abdominal pregnancy in the sac of lesser omentum after in vitro fertilization and embryo transfer (IVF-ET). Methods:A case of abdominal pregnancy in the sac of lesser omentum after IVF-ET was reported in detail and summarized, and its clinical characteristics were analyzed.Results:After two blastocysts were transplanted 23 d, the patient had a sudden left upper abdominal pain. Ultrasound showed no pregnancy sac in utero, left epigastric dysplasia echo, and a small amount of fluid in the abdominal cavity. Puncture of the posterior vaginal fornix was nonclotting, and there was a clear indication of laparoscopic exploration. After cleaning up the blood in the pelvic and abdominal, the gestational tissue of lesser omentum sac was cleared. The patient recovered well after surgery.Conclusion:Abdominal pregnancy in the sac of lesser omentum after IVF-ET is extremely rare and should be treated timely. Laparoscopic surgery is recommended to determinate abdominal pregnancy in the first trimester.
6.Abdominal pregnancy in the sac of lesser omentum after in vitro fertilization and embryo transfer: a case report
Jinghui JIA ; Bing CHEN ; Mengnan YU ; Hongfang WANG ; Ying TONG ; Ying SHI
Chinese Journal of Reproduction and Contraception 2023;43(4):404-406
Objective:To summarize the clinical experience in the diagnosis and treatment of abdominal pregnancy in the sac of lesser omentum after in vitro fertilization and embryo transfer (IVF-ET). Methods:A case of abdominal pregnancy in the sac of lesser omentum after IVF-ET was reported in detail and summarized, and its clinical characteristics were analyzed.Results:After two blastocysts were transplanted 23 d, the patient had a sudden left upper abdominal pain. Ultrasound showed no pregnancy sac in utero, left epigastric dysplasia echo, and a small amount of fluid in the abdominal cavity. Puncture of the posterior vaginal fornix was nonclotting, and there was a clear indication of laparoscopic exploration. After cleaning up the blood in the pelvic and abdominal, the gestational tissue of lesser omentum sac was cleared. The patient recovered well after surgery.Conclusion:Abdominal pregnancy in the sac of lesser omentum after IVF-ET is extremely rare and should be treated timely. Laparoscopic surgery is recommended to determinate abdominal pregnancy in the first trimester.
7.Application value of dynamic monitoring of serum soluble interleukin-2 receptor and stromal cell-derived factor 1α in the evaluation of patients with acute leukemia
Jianlan LI ; Ju QIAO ; Ying YANG ; Lijuan XU ; Zhipeng LIANG ; Hongfang JIA
Journal of Leukemia & Lymphoma 2020;29(4):232-235
Objective:To explore the application value of dynamic monitoring of serum soluble interleukin-2 receptor (sIL-2R) and stromal cell-derived factor 1 α (SDF-1α) in patients with acute leukemia (AL).Methods:A total of 187 patients with AL admitted to the Second Hospital of Shanxi Medical University from March 2017 to August 2018 were selected and 90 healthy subjects at the same period were selected as the healthy controls. The levels of serum sIL-2R and SDF-1α were detected at the initial diagnosis, remission phase and relapse phase, respectively. The clinical value of dynamic monitoring of sIL-2R and SDF-1α was analyzed.Results:Among 187 patients with AL, 135 patients (72.19%) had complete remission (CR) after chemotherapy, 52 patients (27.81%) had partial remission, and 43 patients (31.85%) relapsed. The level of serum sIL-2R at the AL initial diagnosis period was (533±32) U/ml, which was higher than that in the healthy controls [(247±30) U/ml], and the difference was statistically significant ( t = 71.976, P < 0.01); the level of serum SDF-1α at the AL initial diagnosis was (2 968±305) pg/ml, which was higher than that in the healthy controls [(1 358±160) pg/ml], and the difference was statistically significant ( t = 47.043, P < 0.01). The levels of serum sIL-2R [(308±30) U/ml] and SDF-1α [(1 576±184) pg/ml] in the CR phase were lower than those in the initial diagnosis of patients with AL; and the levels of sIL-2R [(599±36) U/ml] and SDF-1α [(2 894±301) pg/ml] in the relapse phase were higher than those in the CR phase of patients with AL (all P < 0.01). Conclusions:The levels of serum sIL-2R and SDF-1α in patients with AL are increased, and there are big differences in the levels of sIL-2R and SDF-1α at the initial diagnosis, remission phase, and relapse phase. Dynamic monitoring of both can provide the data support for early clinical intervention.
8. Diffuse midline glioma of spinal cord with H3K27M-mutation: a clinicopathological analysis
Jingjing YAO ; Donglin MA ; Jing ZHAO ; Ying XIAO ; Hongfang YIN
Chinese Journal of Pathology 2019;48(6):448-452
Objective:
To investigate the role of H3K27M mutant in spinal cord glioma, specifically the correlation between H3K27M mutation and histological grade or prognosis.
Methods:
Twenty-four cases of paraffin-embedded spinal cord glioma tissues and clinical data were collected from November 2014 to August 2016 at the Beijing Tsinghua Changgung Hospital. There were 13 males and 11 females, and the age ranged from 3 to 66 years. All the cases were reviewed histologically, and immunohistochemical H3K27M staining and H3 gene detection were performed. The correlation between H3 gene mutation and histological grading and prognosis of spinal cord gliomas were investigated and relevant literature reviewed.
Results:
Eleven of 24 cases showed H3K27M gene mutation, and was concordant with the result of immunohistochemistry. Gliomas in the mutant group were all high-grade gliomas with mean patients′ age of (30.0±11.5) years, and a male to female ratio of 7:4. Thirteen cases were wild-type, and these included four high-grade gliomas, with mean patients′ age (31.3±22.4) years, and a male to female ratio of 6∶7. The tumors in the mutant group were mainly located in cervical 4-7 and thoracic 11-12 segments, respectively, and the incidence of tumors in the lower thoracic segments (thoracic 11-12) was higher than that in the wild type group. Outcome data were available for all patients. The median survival of mutant group was 19.5 months, but most patients in the wild-type group were alive at the end of the follow-up period.
Conclusion
Gliomas of spinal cord with H3K27M mutation are high-grade and the prognosis of patients is poor.
9. Significance of endoplasmic reticulum stress-associated gene tribbles pseudokinase 3 in the long-term brain injury in developing epileptic rats
Jing ZHANG ; Ying HAN ; Hongfang JIN ; Yang ZHAO ; Qinrui LI ; Jiong QIN
Chinese Journal of Applied Clinical Pediatrics 2019;34(11):854-858
Objective:
To investigate the significance of endoplasmic reticulum stress-associated gene tri-bbles pseudokinase 3 (TRIB3) in the long-term brain injury in rats with developing epilepy.
Methods:
Thirty male SD rats aged 21 days were randomly divided into the control group and the epilepsy group, 15 rats in each group.The rats in the epilepsy group were intraperitoneally injected with kainic acid (10 mg/kg) to induce seizures, while the rats in the control group were injected with the equal volume of 9 g/L saline.The rats in two groups were euthanized at 30 d after kainic acid administration.The damage to the ultrastructure of the cortex were observed by using transmission electron microscopy.Neuronal apoptosis in the cortex of rats was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay.The expression and localization of glucose regulated protein 78 (GRP78), CCAAT/enhancer binding protein-homologous protein (CHOP), TRIB3, and the activation of protein kinase B (AKT) in the cortex were examined by using Western blot analysis and immunohistochemistry.
Results:
Compared with the control group, the different ultrastructural changes were observed in the cortex in the epilepsy group rats.TUNEL assay indicated that the number of apoptosis cells of cortex in the epilepsy group was increased.The protein levels of GRP78 and TRIB3 were upregulated in the cortex of the epileptic rats (1.280±0.272, 1.725±0.570), compared with the control group (1.000±0.000, 1.000±0.000), and the differences were statistically significant (all
10.The evidence-based nursing practice of applying physical cooling methods in the treatment of hyperthermia among in-patients
Yuan LI ; Hongfang YE ; Hong CHU ; Qian SHEN ; Li WAN ; Xiuhua YING ; Xinyue ZHANG
Chinese Journal of Modern Nursing 2019;25(12):1475-1480
?? [Abstract]? Objective? To explore the effect of physical cooling method by the use of external cooling ice-packs for treatment of the in-patients with fever. Methods? In accordance with the model of Australian JBI evidence-based health care, the evidence was applied to the clinical practice after evidence generation, and synthesis. Admitted from November 2017 to June 2018 into the hematology department in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, the hyperpyretic patients with blood disease were divided into two groups-observation group and control group randomly. 68 patients were included in each group. The patients in observation group were given by 0.5 g acetaminophen orally along with ice-packs while the patients in control group were just given by 0.5 g oral acetaminophen. The temperature lowering effect in both groups was compared and studied. Results? In the observation group, patients' temperature was (38.90±0.37)℃ before intervention, (38.85±0.36)℃ in 30 minutes after intervention, (38.63±0.52)℃ in 1 hour and (38.07±0.76)℃ in 2 hours. And those for control group were (38.89±0.39),(38.82±0.40),(38.58± 0.59),(37.90±0.67)℃,respectively. Repeated measurement analysis of variance indicated that patients' temperature showed significant differences at each point of time during the study in both group (Ftime=77.862,P< 0.01). The intervention methods in the two groups did not interact with different point of time (Finteraction=0.728, P>0.05). The effect of intervention in different groups at different point of time was not significant (Fgroups=0.909, P> 0.05). Conclusions? The use of physical cooling method along with medication is proved to be not significant in the patients' temperature control. Health workers should or not select the physical cooling method based on patients' condition and willingness in order to ensure their vital signs' stability and their comfort.

Result Analysis
Print
Save
E-mail