ObjectiveThe Golgi apparatus serves as a central hub in the eukaryotic secretory pathway, responsible for the processing, sorting, and trafficking of proteins and lipids. In mammalian cells, the Golgi typically forms a perinuclear ribbon-like structure composed of laterally connected cisternae stacks.The maintenance of Golgi ribbon structure depends on the balance of membrane flux across multiple intracellular trafficking pathways, yet the specific contributions of distinct trafficking branches to Golgi macroscopic morphology remain elusive. In mammalian cells, the Golgi ribbon is typically organized as a perinuclear, laterally connected structure composed of stacked cisternae, and its integrity is highly dynamic and sensitive to perturbations in membrane trafficking. This study aims to elucidate the role of coat protein complex I (COPI)-mediated retrograde transport in maintaining the Golgi ribbon and to dissect the functional relationship between the transmembrane cargo receptors LEPROT/LEPROTL1 (LEPROTs) and the COPI adaptor GOLPH3. MethodsUsing siRNA interference and gene-deficient cell lines, we selectively perturbed COPI- or adaptor protein complex 1 (AP-1)-mediated trafficking pathways in HeLa cells. To quantitatively evaluate Golgi morphology, we employed a “Golgi Angle”-based measurement to assess its circumferential distribution around the nucleus. The spatial distribution of the Golgi ribbon was quantitatively analyzed using confocal microscopy, while Golgi ultrastructure and vesicle density were examined via transmission electron microscopy. Additionally, the subcellular distribution of COPI components was assessed by immunofluorescence co-localization. ResultsSelective inhibition of COPI retrograde transport significantly induced the circumferential extension of the Golgi ribbon around the nucleus, whereas blocking AP-1-mediated anterograde transport resulted in Golgi compaction, indicating opposing roles. These results suggest that different trafficking branches downstream of ARF1 exert distinct and even antagonistic effects on Golgi morphology. LEPROTs-deficient cells exhibited a Golgi extension phenotype highly consistent with COPI impairment. Furthermore, knockdown of GOLPH3 in a LEPROTs double-knockout background produced a significant additive effect on Golgi extension, suggesting that LEPROTs and GOLPH3 play non-redundant roles in regulating COPI-related trafficking processes. Mechanistically, loss of either LEPROTs or GOLPH3 led to the aberrant accumulation of COPI components at endoplasmic reticulum exit sites, accompanied by a reduction in COPI-like vesicles around the Golgi. This redistribution indicates a defect in COPI recycling between the ER-Golgi interface and the Golgi apparatus. Ultrastructural analysis revealed that Golgi cisternae in defective cells became shorter and thicker while maintaining a stable number of stacks. In parallel, the density of Golgi-associated vesicles was markedly decreased, further supporting an impairment in COPI vesicle formation or budding processes. ConclusionThis study demonstrates that active COPI retrograde transport is a critical factor in restricting the over-connection of the Golgi ribbon and maintaining its compactness. Rather than causing fragmentation, partial disruption of COPI function leads to a distinct morphological outcome characterized by Golgi ribbon extension at the light microscopy level and cisternal remodeling at the ultrastructural level. LEPROTs and GOLPH3 cooperatively promote the recycling and vesiculation of COPI components, thereby imposing a structural constraint on the Golgi periphery. Our findings support a model in which multiple adaptor proteins act in parallel to sustain efficient COPI cycling, thereby maintaining Golgi structural homeostasis. These findings provide new cell biological evidence for the membrane trafficking basis of Golgi morphological homeostasis.

Purpose: This study aimed to evaluate the safety and efficacy of gemcitabine and cisplatin (GP) regimen in combination with immune checkpoint inhibitor sintilimab as neoadjuvant therapy for muscle-invasive bladder cancer (MIBC) patients and the feasibility of the following selective bladder sparing surgery. Materials and Methods: Patients with histopathologically confirmed urothelial carcinoma without distant metastases (T2-4a, N ≤ 1, M0, American Joint Committee of Cancer 8th) and with adequate organ function will be enrolled. The therapeutic regimen was sintilimab 200 mg once on day 8, gemcitabine 1,000 mg/m2 and cisplatin 35 mg/m2 once on days 1 and 8, every 21 days for four cycles. The primary endpoint was pathologic complete response (pCR, pT0N0) rate. The secondary end points were ypT < 2 rate, R0 resection rate, event-free survival, and safety. Results: From May 4, 2020, to May 20, 2023, 55 patients were enrolled. Forty-six patients were evaluated for efficacy. Among the 42 patients who underwent surgery, 16 patients (38.0%) achieved pCR. Thirty-three patients (78.6%) achieved pT < 2. With a median follow-up of 15.7 months, the 1-year event-free survival was 91.3%. Notwithstanding the poor pathological baseline characteristic of a high T3-T4a proportion (39.1%), a promising bladder preservation (including 22 patients transurethral resection of bladder tumor, 5 patients partial cystectomy, and 4 surveillances) rate was achieved (67.4%). The most common grade ≥ 3 treatment-related adverse events was neutropenia (n=15, 27.3%), which was related to chemotherapy. There were no grade 3 immune-related adverse events. Conclusion Neoadjuvant GP plus sintilimab is a promising regimen for MIBC patients, with relatively high pT < 2 rate and triggering the emerging roles for the multi-disciplinary team decision-making for bladder sparing surgery.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

ObjectiveTo determine the effects of tuina at "Weizhong （BL 40）" on mechanical pain hypersensitivity in a rat model of chronic dorsal root ganglion （CCD）， and to preliminarily explore the anatomical distribution characteristics of neural afferents associated with "Weizhong （BL 40）" acupoint and their central projections. MethodsThirty SD rats were randomly assigned to a sham operation group， a model group， and a tuina group， with 10 rats in each group. In the model group and the tuina group， a chronic compression of CCD model was established by implanting stainless-steel rods into the right L4/5 and L3/4 intervertebral foramina. Rats in the sham operation group underwent surgical exposure only. From day 4 to day 28 after modeling， rats in the tuina group received daily manipulation at the right "Weizhong （BL 40）" （pressure： 5 N； frequency： 2 Hz； duration： 10 min/session）. Rats in the sham operation group and the model group were subjected only to fixation. Mechanical withdrawal threshold （MWT） was assessed before modeling and on day 3， 7， 14， 21， and 28 after modeling. On day 29 after modeling， pseudorabies virus （PRV）， a retrograde trans-multisynaptic tracer， was injected into the right "Weizhong （BL 40）" acupoint in rats from the model group. After 120 h， rats were perfusion-fixed through the heart， and local tissues surrounding the "Weizhong （BL 40）" acupoint， corresponding lumbar dorsal root ganglia， lumbar spinal cord， and brain tissues were collected for fluorescence imaging of PRV-positive signals. ResultsCompared to the sham operation group， rats in the model group and the tuina group showed significantly reduced MWT on day 3， 7， 14， 21， and 28 after modeling （P＜0.05）. Compared to the model group， rats in the tuina group exhibited significantly increased MWT on day 21 and 28 after modeling （P＜0.05）. Fluorescence imaging revealed PRV-positive signals in the deep fascial layer of local muscle tissue at "Weizhong （BL 40）"， the L4 and L5 dorsal root ganglia， the dorsal horn and central canal region of the spinal cord， as well as the primary somatosensory cortex （S1）， secondary motor cortex （M2）， and anterior cingulate cortex （ACC）. Among the examined cortical regions， M2 contained the largest number of PRV-positive neurons. ConclusionTuina at "Weizhong （BL 40）" alleviates mechanical hyperalgesia in rats with the chronic compression of CCD model. A traceable neuroanatomical connection exists between the linking deep tissues within "Weizhong （BL 40）" region and the corresponding lumbar dorsal root ganglia， the spinal dorsal horn， and specific cortical areas.

Objective To investigate the effects of plateau hypoxia on the growth and tumor microenvironment of colorectal carcinoma in vivo.Methods A total of 16 male BALB/C mice(6 weeks old,weight 18-20 g)were randomly divided into plateau hypoxic group and plain normoxic group,with 8 mice in each group,while 14 male C57BL/6 mice were grouped in same way,with 7 mice in each group.The mice in the plateau hypoxic group were housed in a low-pressure oxygen(10%)chamber to simulate an altitude of approximately 5 600 m,while the mice of the other group was maintained in SPF-grade normal atmospheric conditions(21%oxygen,at an altitude of about 300 m).Colorectal tumor MC38 cells and colon adenocarcinoma CT26 cells were subcutaneously implanted into C57BL/6 mice and BALB/C mice,respectively to construct subcutaneous tumor-bearing mouse models.Then the tumor size and weight were measured in 4 groups of mice.After the tumor tissues,spleen and blood samples were collected in the C57BL/6 mice.Flow cytometry was used to determine the percentages of macrophages,T lymphocytes,IFN-γ+T lymphocytes,and myeloid-derived suppressor cells(MDSC).The differences in these immune cells were compared between the cells from the plateau hypoxic group and those from the plain normoxic group.Results The weight of subcutaneous tumor mass was significantly inhibited in both C57BL/6 and BALB/C mice from the plateau hypoxic group than those from the 2 plain normoxic groups(0.17 vs 0.09 g,1.38 vs 0.51 g,P<0.01).When compared with the immune cells from the tumor mass of the plain normoxic C57BL/6 mice,the percentage of M2-type macrophages was reduced in the tumor tissue from the plateau hypoxic mice(22.13%vs 15.90%,P<0.05),so was that of MDSC(2.06%vs 1.05%,P<0.01),particularly in the monocytic(M)-MDSC subgroup(60.97%vs 41.13%,P<0.01).While,no significant change was observed in the proportion of the polymorphonuclear(PMN)-MDSC subgroup(10.97%vs 9.70%,P>0.05).Additionally,the percentage of CD4+T cells was significantly reduced(48.70%vs 41.93%,P<0.05),whereas that of CD8+T cells was obviously increased(41.25%vs 51.18%,P<0.05),along with a notable rise in the proportions of IFN-γ+T,IFN-γ+CD4+T and IFN-γ+CD8+T cells(28.58%vs 59.65%,23.33%vs 53.65%,36.9%vs 66.48%,P<0.01).However,between the peripheral blood samples of the 2 groups of C57BL/6 mice,the proportions of M1-type macrophages and CD4+T cells were reduced(84.98%vs 78.43%,5.86%vs 4.01%,P<0.01),and those of MDSC and PMN-MDC were increased(4.47%vs 16.43%,36.56%vs 62.97%,P<0.01).In the spleen tissues,notable decreases were observed in the proportions of CD8+T cells and IFN-γ+CD8+T cells between the 2 groups(33.05%vs 27.68%,5.13%vs 1.58%,P<0.01).Conclusion Plateau hypoxia improves the immune response within the tumor microenvironment,and inhibits subcutaneous tumor growth of colorectal cancer,but suppresses systemic immune response.

Aim To investigate the therapeutic effects of a newly developed Tadalafil tablet on pulmonary fi-brosis induced by paraquat(PQ)in rats,as well as its impact on histone acetylation levels in epithelial cells.Methods SD rats were randomly divided into four groups:the control group(control),the model group(PQ),the Tadalafil new tablet treatment group(N-Tad,1 mg·kg-1),and the positive control drug treatment group(Cialis,5 mg·kg-1).The model group and treatment group rats were intraperitoneally injected with PQ(30 mg·kg-1).Two hours after the initial treatment,the rats in the treatment group re-ceived N-Tad or Cialis via gavage,while the control and model groups were administered an equal volume of physiological saline by gavage once daily for 28 days.The weight gain rate and lung tissue index for each group of rats were calculated.Additionally,the effects of N-Tad treatment on lung tissue structural damage and collagen deposition in rats with PQ-in-duced pulmonary fibrosis were observed using HE stai-ning,Masson trichrome staining,and immunohisto-chemical techniques.By employing the Western blot technique,the effects of Tadalafil intervention on the expression of the epithelial marker E-cadherin(E-Cad),the stromal marker fibronectin(Fn),and the histone acetylation marker acetylated histones(Ac-his-tones)in A549 cells were observed.Results Com-pared to the control group,rats with PQ-induced pul-monary fibrosis exhibited a significant decrease in the rate of body weight growth,an increase in lung tissue index(P＜0.05),and a notable increase in the expression and distribution of the fibrosis marker alpha-smooth muscle actin(α-SMA)in lung tissue.The structure of the lung tissue was disrupted,accompanied by the deposition of interstitial collagen fibers.Both N-Tad and Cialis treatments could significantly enhance the rate of weight gain,decrease the lung tissue index,inhibit the expression of α-SMA,and reduce the depo-sition of interstitial collagen in the lung tissue of rats with pulmonary fibrosis.Notably,low-dose N-Tad treatment was comparable to high-dose Cialis treat-ment.At the cellular level,Tadalafil significantly in-hibited the high expression of Fn induced by transfor-ming growth factor beta 1(TGF-β1)in A549 cells.It also upregulated the expression of E-cadherin and sig-nificantly increased the levels of acetylated histones(P＜0.05).Conclusions N-Tad promotes histone acetylation in alveolar epithelial cells,significantly in-hibits epithelial-mesenchymal transition,increases E-cadherin expression,and improves lung tissue structur-al damage and collagen deposition caused by PQ.Ad-ditionally,it offers the advantage of a lower effective dose compared to Cialis,providing a new option for the treatment of pulmonary fibrosis.

Objective To examine and compare the effect of personalized progressive aerobic-exercise and resistance-training prescriptions on anxiety of undergraduates.Methods This was a randomized controlled trial.Sixty-six undergraduates with anxiety were recruited and randomized into an aerobic ex-ercise(AE)group,a resistance exercise(RE)group and a control group,each of 22.The aerobic and resistance exercise groups underwent 12-week aerobic and resistance exercise respectively,while the control group only received health education.Before as well as after 4-,8-and 12-week interven-tion,and 4 weeks after the intervention,all groups were evaluated using Self-Rating Anxiety Scale(SAS)and Chinese College Students Mental Health Scale--Anxiety Subscale(CCSMHS-AS).More-over,before and 16 weeks after the intervention,all groups were assessed physical activity(PA)us-ing the International Physical Activity Questionnaire-Short Form(IPAQ-SF).Results The average SAS scores of the AE and RE groups decreased significantly from 6.95±6.32 and 56.41±5.45 before the intervention to 38.29±5.82 and 41.18±7.51 after 12-week exercise,while the average CC-SMHS-AS score decreased significantly from 20.00±5.66 and 19.41±3.70,to 13.18±4.81 and 14.32±4.16 during the same period of time(P<0.01 for all).Four weeks after the intervention,the SAS score of the AE group was significantly higher than 4 weeks earlier(49.18±11.84 vs.38.29±5.82,P<0.01),while that of the RE group increased without significant differences compared with 4 weeks earlier(42.50±9.57 vs.41.18±7.51,P>0.05),with the value of both groups significantly lower than right after the intervention(P<0.01,P<0.05).In the control group,the SAS score de-creased significantly from 55.73±5.27 before the intervention to 47.09±5.55 right after the interven-tion,and further to 46.95±9.70 4 weeks later(P<0.05),but no significant differences were ob-served in the CCSMHS-AS score(P>0.05).Meanwhile,right after the intervention,the average SAS scores of the AE and RE groups were significantly lower than the control group(P<0.01,P<0.05),without significant differences among the three groups 4 weeks after the intervention(P>0.05).The CC-SMHS-AS scores of AE group right after and 4 weeks after the intervention were significantly higher than the control group(P<0.01),but no significant differences were found in it between either the AE and RE group,or the RE and control group(P>0.05).Besides,the PA levels of the AE and RE groups 4 weeks after the intervention were significantly higher than before the intervention,while no significant changes were observed in the PA level of the control group(P>0.05).Conclusion Twelve-week personalized progressive aerobic-exercise and resistance-training prescriptions both result in a similar effect on relieving anxiety and improving spontaneous PA of college students.However,the prognosis of aerobic exercise is poorer than the other.

Aim To investigate the possible mechanism of the myocardial protective effect of Danzhi Jiangtang capsules(DJC)on db/db mice based on NLRP3 in-flammasome-mediated pyroptosis.Methods The db/db mice were randomly divided into the model group,DJC low,medium,and high dose groups,and the met-formin group,and the db/m mice were taken as the blank group.The administration lasted for eightweeks.At the end of drug administration,blood glucose,blood lipids,cardiac enzymes and inflammatory factors were detected in each group of mice.HE and Masson stai-ning was performed to observe the morphology and fi-brosis of myocardial tissue.TUNEL staining was per-formed to detect apoptosis.RT-qPCR was performed to detect the mRNA expression of ANP,BNP and β-MHC,and Western blot was performed to detect the protein expression of NLRP3,ASC,caspase-1,cleaved-caspase-1,GSDMD and GSDMD-NT in myocardial tis-sue.Results DJC could alleviate myocardial patho-logical damage,reduce collagen deposition and apopto-sis,reduce the levels of blood glucose,blood lipid,myo-cardial enzyme and inflammatory factors in db/db mice.DJC could reduce the mRNA expressions of ANP,BNP and β-MHC,and the protein expressions of NLRP3,ASC,caspase-1,cleavedcaspase-1,GSDMD and GSDMD-NT in myocardial tissues.Conclusion DJC attenuates myocardial injury in db/db mice,prob-ably by inhibiting the activation of NLRP3 inflamma-somes,attenuating cardiomyocyte pyroptosis,and amel-iorating the inflammatory state.

Aim To investigate the therapeutic effects of a newly developed Tadalafil tablet on pulmonary fi-brosis induced by paraquat(PQ)in rats,as well as its impact on histone acetylation levels in epithelial cells.Methods SD rats were randomly divided into four groups:the control group(control),the model group(PQ),the Tadalafil new tablet treatment group(N-Tad,1 mg·kg-1),and the positive control drug treatment group(Cialis,5 mg·kg-1).The model group and treatment group rats were intraperitoneally injected with PQ(30 mg·kg-1).Two hours after the initial treatment,the rats in the treatment group re-ceived N-Tad or Cialis via gavage,while the control and model groups were administered an equal volume of physiological saline by gavage once daily for 28 days.The weight gain rate and lung tissue index for each group of rats were calculated.Additionally,the effects of N-Tad treatment on lung tissue structural damage and collagen deposition in rats with PQ-in-duced pulmonary fibrosis were observed using HE stai-ning,Masson trichrome staining,and immunohisto-chemical techniques.By employing the Western blot technique,the effects of Tadalafil intervention on the expression of the epithelial marker E-cadherin(E-Cad),the stromal marker fibronectin(Fn),and the histone acetylation marker acetylated histones(Ac-his-tones)in A549 cells were observed.Results Com-pared to the control group,rats with PQ-induced pul-monary fibrosis exhibited a significant decrease in the rate of body weight growth,an increase in lung tissue index(P＜0.05),and a notable increase in the expression and distribution of the fibrosis marker alpha-smooth muscle actin(α-SMA)in lung tissue.The structure of the lung tissue was disrupted,accompanied by the deposition of interstitial collagen fibers.Both N-Tad and Cialis treatments could significantly enhance the rate of weight gain,decrease the lung tissue index,inhibit the expression of α-SMA,and reduce the depo-sition of interstitial collagen in the lung tissue of rats with pulmonary fibrosis.Notably,low-dose N-Tad treatment was comparable to high-dose Cialis treat-ment.At the cellular level,Tadalafil significantly in-hibited the high expression of Fn induced by transfor-ming growth factor beta 1(TGF-β1)in A549 cells.It also upregulated the expression of E-cadherin and sig-nificantly increased the levels of acetylated histones(P＜0.05).Conclusions N-Tad promotes histone acetylation in alveolar epithelial cells,significantly in-hibits epithelial-mesenchymal transition,increases E-cadherin expression,and improves lung tissue structur-al damage and collagen deposition caused by PQ.Ad-ditionally,it offers the advantage of a lower effective dose compared to Cialis,providing a new option for the treatment of pulmonary fibrosis.

Objective To investigate the effect of oxymatrine on the brain damage of rats with acute carbon monoxide(CO)poisoning through the sirtuin 1(SIRT1)/forkhead box protein O1(FOXO1)signaling pathway.Methods SD rats were used to establish an acute CO poisoning rat model,after intervention with low,medium,and high doses of oxymatrine and edaravone,the cognitive function of the rats was tested using shuttle box experiments to screen for the optimal dosage of oxymatrine.Construct a rat model of acute CO poisoning againand randomly divide it into five groups:control group,model group,oxymatrine group,edaravone group,EX527(SIRT1 inhibitor)group,and oxymatrine+EX527 group,the model group and drug intervention groupinhaled CO gas to construct acute CO poisoning rat model.After drug intervention,the shuttle box experiment was used to detect the cognitive function in rats,the step-through latency(STL)and the total time spent in the dark chamber(TDC)of each group were compared;fluorescent probe was performed to measure the mitochondrial membrane potential of rat brain tissue;TUNEL staining was performed to detect the apoptosis rate of hippocampal neurons in the rat brain;the kit was performed to determine the levels of serum inflammatory factors and the oxidative stress factor;immunoblotting and immunoprecipitation experimentswas performed to determine the expression of SIRT1/FOXO1 pathway protein.Results Compared with the control group,the STL,brain mitochondrial membrane potential,serum SOD level,and brain tissue SIRT1 protein expression in the model group were significantly reduced(P＜0.05),and the TDC,neuron apoptosis rate,serum ROS,PGE2,TNF-α,IL-18 and MDA levels,and brain tissue acely-FOXO1/FOXO1 were significantly increased(P＜0.05).Compared with the model group and the oxymatrine+EX527 group,the STL,brain mitochondrial membrane potential,serum SOD level,and brain tissue SIRT1 protein expression were all increased in the oxymatrine group(P＜0.05),and the TDC,neuron apoptosis rate,serum ROS,PGE2,TNF-α,IL-18 and MDA levels,and brain tissue acely-FOXO1/FOXO1 were all decreased(P＜0.05);the trend of changes in various indicators in the EX527 group is opposite to that of the oxymatrine group(P＜0.05).There was no significant change in the levels of various indicators between the edaravone group and the high-dose oxymatrine group(P＞0.05).Conclusion Oxymatrine can activate SIRT1/FOXO1 signal to reduce inflammation and oxidative stress in rats,inhibit hippocampal neuronal apoptosis,repair brain mitochondrial function,enhance cognitive ability of rats,and improve brain damage of acute CO poisoning rats.