1.Clinical observation of 25G+minimally invasive vitrectomy in the treatment of retinal arterial macroaneurysm
Nan ZHAO ; Tongtong LIU ; Yige XIA ; Haohao LU ; Yanhui HAN
International Eye Science 2025;25(7):1191-1194
AIM:To observe the therapeutic efficacy of 25G+minimally invasive vitrectomy for retinal arterial macroaneurysm.METHODS:Totally 40 patients(40 eyes)who admitted to Jinan Mingshui Eye Hospital from January 2021 to May 2024 and with vitreous hemorrhage or dense premacular hemorrhage in the macular area caused by retinal arterial macroaneurysm, underwent 25G+minimally invasive vitrectomy. Preoperative and postoperative best-corrected visual acuity(BCVA), complications, and special cases were analyzed.RESULTS: The general patient data aligned with previous literature reports. The postoperative BCVA was significantly improved(t=9.72, P<0.01), and no significant serious surgical complications were observed. Notably, intraoperative findings revealed secondary macular holes in 3 eyes, resulting in poor visual prognosis.CONCLUSION: For vitreous hemorrhage or dense premacular hemorrhage caused by retinal arterial macroaneurysm, 25G+ minimally invasive vitrectomy is a safe and effective treatment. Visual prognosis was excluded for secondary macular holes.
2.Effect of neferine on diabetic nephropathy by regulating SDF-1/CXCR4 signal pathway
Ying WANG ; Lifeng PING ; Tongtong LIU ; Shanshan LIU ; Lei LIU
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(2):183-195
Objective·To investigate the effect of neferine(Nef)on renal tissues of diabetic nephropathy(DN)rats and its related mechanism.Methods·DN model rats were constructed by feeding high-fat diet combined with intraperitoneal injection of streptozotocin,and the successfully constructed rats were randomly divided into DN group,Nef(low,medium and high)dose groups and Nef high-dose+pathway antagonist(AMD3100)group,with 10 rats in each group.At the same time,10 common rats were selected as the normal group.The levels of fasting blood glucose(FBG),24 h urinary protein,serum glycosylated hemoglobin(HbA1c),serum creatinine(Scr),blood urea nitrogen(BUN)and renal index of rats in the six groups were measured.Hematoxylin-eosin(H-E)and Masson staining were used to observe the pathological changes of renal tissues.The content of malondialdehyde(MDA)in renal tissues was determined by thiobarbituric acid(TBA)method,and the activities of superoxide dismutase(SOD)and catalase(CAT)in renal tissues were determined by water soluble tetrazolium(WST-1)method and ammonium molybdate method,respectively.The mRNA and protein expressions of stromal cell-derived factor-1(SDF-1)and CXC chemokine receptor 4(CXCR4)in renal tissues were detected by quantitative real-time PCR(qPCR)and Western blotting,respectively.Rat renal tubular epithelium cells NRK-52E were induced by high glucose(30 mmol/L glucose)to establish DN cell model.The cells were divided into control group,high glucose(HG)group,HG+Nef(low,medium and high)dose(i.e.HG+Nef-L,M and H)group,and HG+Nef-H +AMD3100 group.SOD and CAT activities were detected by WST-1 method and ammonium molybdate method,respectively.MDA content was detected by TBA method.The mRNA and protein expressions of SDF-1 and CXCR4 were detected by qPCR and Western blotting,respectively.CCK-8 method and flow cytometry were used to detect cell viability and apoptosis rate,respecti-vely.Results·Compared with the DN group,the levels of FBG,24 h urinary protein,HbA1c,Scr,BUN,renal index and MDA content in Nef(low,medium and high)dose groups and Nef high-dose+AMD3100 group were decreased,the mRNA and protein expressions of SDF-1 and CXCR4 were increased,and the activities of SOD and CAT were increased(all P<0.05).The degree of pathological damage and fibrosis of renal tissues was reduced;all of the above changes were dose-dependent.AMD3100 could weaken the renal protective effect of high-dose Nef on DN rats.Compared with the HG group,NRK-52E cell viability,SOD and CAT activities,and the mRNA and protein expressions of SDF-1 and CXCR4 were increased in HG+Nef-L,M and H groups,while apoptosis rate and MDA content were decreased(all P<0.05).AMD3100 could reverse the protective effect of Nef-H on NRK-52E cell damage.Conclusion·Nef may control blood glucose levels on DN rats and improve antioxidant capacity by activating the SDF-1/CXCR4 signal pathway,playing a renal protective role.
3.Effect of LncRNA SNHG1 on homocysteine-induced podocytespyrophosis
Zhenghao ZHANG ; Fang MA ; Qing ZHANG ; Tongtong XIA ; Honglin LIU ; Zhigang BAI ; Guanjun LU ; Jingwen ZHANG ; Hongjian PENG ; Yideng JIANG ; Shengchao MA
The Journal of Practical Medicine 2024;40(4):476-482
Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte.Methods Cbs+/-mice were randomly divided into two groups:a normal diet group(ND)and a high me-thionine diet group(HMD).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocytes were cultured in vitro,and then assigned into a control group(Control,0 μmol/L Hcy)and a homocysteine intervention group(Hcy,80 μmol/L Hcy).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocyion group(OE-NC + Hcy)and the lncSNHG1 overexpression + homocysteine intervention group(OE-SNHG1 + Hcy)were also established.After 48 hours of intervention,Real-time fluorescence quantita-tive PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention.Western blot was used to detect the expressions of Caspase-1,Cleaved Caspase-3 and NLRP3.Immunofluorescence was used to de-tect the expression levels of GSDMD and GSDMD-N.ELISA was used to detect the contents of IL-1β and IL-18.Results(1)In the animal experiments,the expression levels of pyroptosis-related proteins Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the HMD group compared with the ND group.(2)In the cellular experiments,the expression levels of Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the Hcy group compared with the Control group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased as well.(3)In the cellular experiments,the expres-sion of lncSNHG1 was increased in the Hcy group compared with the control group.After transduction with lnc-SNHG1 lentivirus,the expression of lncSNHG1 was increased in the OE-SNHG1 group,compared with the control group and the OE-NC group.(4)In the cellular experiments,the expressions of pyroptosis-related proteins Cas-pase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were increased compared with the OE-NC+Hcy group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group.Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.
4.Co-loaded with miR-145/CPT nanoparticles for targeted delivery and magnetic resonance imaging in hepatocellular carcinoma
Jing RONG ; Tongtong LIU ; Xiujuan YIN ; Lei ZHANG ; Xiao WANG
Acta Universitatis Medicinalis Anhui 2024;59(9):1501-1508
Objective To develop liver cancer-targeted nanoparticles(LA-CMGL)co-loaded with miR-145/camp-tothecin(CPT)and assess their targeting specificity,combined anti-tumor effects,and magnetic resonance imaging efficacy.Methods Laser scanning confocal microscopy and flow cytometry were utilized to evaluate the targeted uptake of lactobionic acid-modified and unmodified nanoparticles by HepG2 cells and HepaRG cells;quantitative polymerase chain reaction(qPCR)was employed to assess miR-145 content in tumor cells and tissues.The cytotox-icity of CPT,LA-CPT-LNPs and LA-CMGL on HepG2 cells were assessed using the CCK-8 assay.qPCR was also used to evaluate the effect of CPT,LA-CPT-LNPs and LA-CMGL on apoptosis of HepG2 cells.MRI was performed to measure the relaxation rate of LA-CMGL and evaluate its targeting imaging effect on liver cancer cells.Results The uptake rate of LA-CMGL by HepG2 cells surpassed that of CMGL significantly.The relative miR-145 content in liver cancer cells and mouse liver cancer tissues in the LA-CMGL group was markedly higher compared to free miR-145 and CMGL groups(P<0.001).The apoptosis rate of HepG2 cells in the LA-CMGL group exceeded that in the CMGL group and CPT group(P<0.01).At the same Gd3+concentration,the relaxation rate of LA-CMGL significantly surpassed that of Gd-DOTA,and the MRI signal of LA-CMGL in HepG2 cells markedly increased com-pared to CMGL and Gd-DOTA.Conclusion LA-CMGL exhibits promising liver cancer-targeted delivery,com-bined anti-tumor effects,and MRI liver cancer cell-targeted imaging,offering a novel avenue for combined drug/gene therapy for liver cancer.
5.Effect of Mahuang Xixin Fuzitang on Migration of Dendritic Cells in Mice by Regulating Rho/ROCK Signaling Pathway
Shuyue FAN ; Luwei WU ; Tongtong SUN ; Yuying NI ; Jing GAO ; Ying CUI ; Min LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):35-42
ObjectiveTo investigate the inhibitory effect of Mahuang Xixin Fuzitang on the migration of dendritic cells (DCs) in mice and its underlying mechanism. MethodMouse bone marrow-derived DCs were isolated and cultured. The morphological changes of the cells at different stages were observed under a microscope, and the CD11c+ proportion was detected by flow cytometry to identify DC purity. Cells were treated with Mahuang Xixin Fuzitang (1, 2, 5, 10, 20, 40, 50, 100 g·L-1) for 24 hours, and the effect of Mahuang Xixin Fuzitang on cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay to determine the appropriate concentrations for treatment. After modeling by lipopolysaccharide (LPS) induction, DCs were divided into a blank group, a model group, and Mahuang Xixin Fuzitang groups (2, 4, 8 g·L-1). The expression of surface molecules CD80, CD86, and major histocompatibility complex-Ⅱ (MHC-Ⅱ) were detected by flow cytometry. Transwell chamber assay was used to observe cell migration. The levels of chemokine C-C-primitive receptor 7 (CCR7) and chemokine C-X-C-primitive receptor 4 (CXCR4) on the cell surface were detected by enzyme-linked immunosorbent assay (ELISA). The expression of filamentous actin (F-actin) in the cell microfilament cytoskeleton was detected by immunofluorescence (IF) staining. Real-time quantitative polymerase chain reaction (Real-time PCR) was employed to determine the mRNA expression levels of Ras homolog family member A (RhoA) and Rho-associated coiled-coil containing protein kinase 1 (ROCK1). Western blot analysis was performed to detect the protein expression of RhoA and ROCK1. ResultCompared with the blank group, the model group exhibited significantly higher expression levels of CD80, CD86, and MHC-Ⅱ (P<0.01), a significantly increased number of cells migrating to the lower chamber (P<0.01), and significantly elevated levels of CCR7 and CXCR4 (P<0.05, P<0.01). Additionally, F-actin expression was significantly increased (P<0.01), and both RhoA and ROCK1 mRNA and protein expression levels were significantly higher (P<0.05, P<0.01). Compared with the model group, treatment with Mahuang Xixin Fuzitang (2, 4, 8 g·L-1) for 24 hours resulted in significantly lower expression levels of CD80, CD86, and MHC-Ⅱ (P<0.01), a significantly reduced number of cells migrating to the lower chamber (P<0.05), and significantly decreased levels of CCR7 and CXCR4 (P<0.05, P<0.01). Furthermore, F-actin expression was significantly reduced (P<0.01), and both RhoA and ROCK1 mRNA and protein expression levels were significantly decreased (P<0.05, P<0.01). ConclusionMahuang Xixin Fuzitang can inhibit the migration of DCs in mice, and its mechanism of action may be related to reducing the activity of the Rho/ROCK signaling pathway, thereby affecting changes in the cell cytoskeleton.
6.The clinical application value of MR defecography in the diagnosis of pelvic organ prolapse with concomitant stress urinary incontinence
Min LI ; Sumei WANG ; Tongtong LIU ; Xiangnan LI
Journal of Practical Radiology 2024;40(10):1653-1657
Objective To evaluate the diagnostic efficacy of magnetic resonance defecography(MRD)in moderate or severe pelvic organ prolapse(POP)complicated with stress urinary incontinence(SUI).Methods Patients with grade Ⅱ or above POP were prospectively selected,and divided into POP combined with SUI group and isolated POP group.The MRD parameters of pelvic dysfunction between groups were analyzed to clarify the clinical diagnostic value and to evaluate the diagnostic efficacy of MRD in POP combined with SUI.Results This study enrolled 99 POP patients,42 patients in isolated POP group,and 57 patients in POP combined with SUI group.The comparison between groups showed there were significant differences in the function of the urethra and bladder,including the position of the urethro-vesical junction(1.61 cm±1.03 cm vs 2.13 cm±0.71 cm),the position of the posterior bladder wall(4.13 cm±2.46 cm vs 5.23 cm±2.21 cm),the urethra angle(94.22°±35.62°vs 114.28°±35.55°),the length of the closed urethra(2.28 cm±0.90 cm vs 2.15 cm±0.65 cm),and bladder funneling(23.81%vs 45.61%),all the differences were statistically significant(P<0.05).The area under the curve(AUC)of multiple parameters MRD in the diagnosis of SUI was 0.73.The AUC in diagnosis of SUI by MRD combined with clinical history was 0.95.Conclusion POP combined with SUI has characteristic MRD signs.MRD technology can assist clinicians and improve the preoperative diagnosis rate of moderate and severe POP combined with SUI.
7.The effect of wza gene deletion in Klebsiella pneumoniae on capsule formation and bacteriophage sensitivity
Zheng FAN ; Hongbo LIU ; Yuchen CHEN ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(7):992-997
Objective:To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods:The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ wza were detected by measuring the optical density OD 600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δ wza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ wza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ wza mutant strain. Results:The PCR results revealed that the Δ wza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ wza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ wza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ wza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups ( t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion:Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.
8.Expression, purification and functional validation of phage depolymerase from hypervirulent Klebsiella pneumoniae serotype K1
Zheng FAN ; Yuchen CHEN ; Hongbo LIU ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(9):1348-1353
Objective:To express and purify the phage depolymerase from hypervirulent Klebsiella pneumoniae (hv Kp) serotype K1 and validate its function. Methods:Phage that infected serotype K1-type hv Kp was isolated from hospital sewage. The biology and morphology of the phage were determined by plaque assay and transmission electron microscopy. The whole genome of the phage was sequenced by the Illumina HiSeq 2500 platform. The presence of depolymerase was determined by observing the plaque halo. Bioinformatic analysis and prokaryotic protein expression system were further used to predict and identify phage depolymerase. The depolymerase gene fragment was obtained by PCR and cloned into the pET28a expression vector, and the expression and purification of the depolymerase were completed in strain BL21. The depolymerase activities on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates were detected by plaque assay and low-speed centrifugation assay. Results:A lytic phage (phiA2) that infected serotype K1-type hv Kp clinical isolate was isolated from hospital sewage. It was typical of the Caudovirales order and Autographiviridae family, and its whole genome was 43 526 bp in length and contained 51 coding domain sequences. The phage phiA2-derived depolymerase phiA2-dep was predicted, expressed and purified. The plaque assay and low-speed centrifugation assay indicated that the depolymerase phiA2-dep had good lytic activity on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates. Conclusion:Depolymerase phiA2-dep can specifically degrade the capsular polysaccharide of serotype K1-type hv Kp, which has potential application value in treating bacterial infection.
9.Analysis of clinical manifestations and prognosis of primary systemic light chain amyloidosis with liver involvement
Tongtong QIAO ; Yang LIU ; Nan PENG ; Lizhong GONG ; Xuelin DOU ; Lei WEN ; Jin LU
Chinese Journal of Hepatology 2024;32(3):222-227
Objective:To summarize the clinical manifestations and prognostic factors of patients with hepatic amyloidosis in a single center.Methods:The clinical data of 28 primary systemic light chain amyloidosis cases with liver involvement in our center from October 2012 to January 2023 were retrospectively analyzed. The main clinical manifestations and prognostic factors were studied. Statistical analysis were performed using the χ 2 test, Fisher's exact test, Wilcoxon rank test, or Kaplan-Meier survival curve log-rank test according to the different data. Results:The main clinical manifestations of patients with liver involvement were abdominal distension, hepatomegaly, and edema. CD56 and chemokine receptor 4 protein expression accounted for 52% (13/25) and 56% (14/25). 64.3% (9/14) patients were combined with t (11,14), and 21.4% (3/14) patients were positive for 1q21 (+), and no patients were detected with del(17p). Univariate analysis showed that Mayo 2004 and 2012 stages and total bilirubin (TBil) ≥34.2 μmol/L were associated with progression-free survival and overall survival. The median progression-free survival and overall survival were significantly inferior in patients with TBil≥34.2μmol/L group (0.178 years, 0.195 years) than with the TBil<34.2μmol/L group (0.750 years, 3.586 years) ( P ?0.05). Conclusion:Mayo stage and hyperbilirubinemia are inferior prognostic factors for patients with primary systemic light chain amyloidosis accompanied with liver involvement.
10.Perceived social support of medical staffs working in community health service institutions in Shanghai Minhang District
Qi SHEN ; Shuai LIU ; Bin CHEN ; Yonghong MU ; Tongtong WEI
Chinese Journal of General Practitioners 2024;23(7):694-701
Objective:To survey the perceived social support status and influencing factors of medical staffs working in community health service institutions in Shanghai Minhang District .Methods:The study was a cross-sectional study. From September 2022 to January 2023, a survey on the status of perceived social support was conducted among 181 medical staffs working in community health service institutions in Shanghai Minhang District. The self-designed general information questionnaire, perceived social support scale, general self-efficacy scale, quality of professional life scale, and trait coping style scale were used in the survey. The effects of demographic characteristics, general self-efficacy, empathy fatigue and trait coping style on perceived social support status of community medical workers were analyzed.Results:A total of 181 questionnairs were distributed and 178 valid ones returned with a valid recovery rate of 98.3%. In three dimensions of perceived social support, family support scored (17.11±8.87), friend support scored (17.08±8.41), other support scored (19.89±4.09), and the total score of understanding social support was (60.99±11.71). There were 2 cases of low support status (1.12%), 81 cases of intermediate support status (45.51%), and 95 cases of high support status (53.37%). Univariate analysis showed that there were significant differences in perceived social support among participants with different ages, marriage status, majors, professional titles, job satisfaction, positive or negative coping styles, general self-efficacy, and empathy fatigue ( P<0.05). Multivariate logistic regression analysis showed that people aged 40 and above had a higher degree of perceived social support than those under 40 ( OR=2.792, 95% CI: 1.051-7.418, P<0.05); compared with public health professionals, clinical medicine, nursing and other medical technique professionals had a higher degree of perceived social support ( OR=4.024, 95% CI: 1.479-10.953; OR=2.941, 95% CI: 1.031-8.393; OR=3.996, 95% CI: 0.543-29.409, all P<0.05); compared with those who were less satisfied with their self-assessment work, participants who were more satisfied or generally satisfied with their self-assessment work had lower perceived social support ( OR=0.200, 95% CI: 0.051-0.781; OR=0.320, 95% CI: 0.146-0.701, all P<0.05); compared with those with mild and moderate empathy fatigue, participants with high empathy fatigue had lower perceived social support ( OR=0.406, 95% CI: 0.182-0.909, P<0.05). Except supporting primary and secondary traumatic stress, the total score of perceived social support and its dimensions were positively correlated with empathy satisfaction; and negatively correlated with job burnout and secondary traumatic stress. Regression analysis shows that empathy satisfaction had a positive predictive effect on overall scores of perceived social support and its dimensions of friend support and other support capacity. Conclusions:The social support status of community healthcare workers in Shanghai Minhang district is generally acceptable. However, it is important to pay special attention to the social support status of those who are under 40 years old, have a background in public health, are satisfied with their work, and experience a high level of empathy fatigue. Measures should be implemented to specifically enhance empathy satisfaction among healthcare workers; timely guidance and intervention should be provided accordingly.


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