Objective:To investigate the effect of ginsenoside octanoate(Rh2-O)on inducing immunogenic cell death in hepa-tocellular carcinoma cells and its molecular mechanism.Methods:Effects of ginsenoside caprylate(Rh2-O)and autophagy inhibitor 3-MA on the activity of hepatocellular carcinoma cells were detected by CCK-8 assay.The effect of Rh2-O on CRT membrane eversion in Hepa1-6 cells were detected by immunofluorescence assay.Rh2-O treated mouse hepatocellular carcinoma cells were used to pre-pare a tumor vaccine for in vivo vaccination experiments in mice.Extracellular ATP levels were detected in real-time.The expression of autophagy-related genes and proteins were measured by real-time fluorescence PCR and Western blot,and the mitochondrial morphol-ogy and co-localization with autophagy proteins were observed by laser confocal microscopy.Results:Rh2-O showed strong cytotoxicity to Hepa1-6 cells[cell viability:(58.54±3.56)%]at a concentration of 150 μmol/L,and a large amount of CRT was observed on the surface of the cell membrane.The tumor emergence rate was 36.36%in the vaccinated group and 100%in the control group.The tumor vaccine prepared by Rh2-O effectively protected mice from the same type of tumor attack;Rh2-O induced an increase in the level of cellular secreted ATP(P<0.05),the mRNA of autophagy-related genes ATG3,p62,LC3 expression levels and autophagy-associated proteins LC3A and LC3B expression levels were increased(P<0.05),and co-localization of mitochondria with autophagy proteins was significantly increased(P<0.05).In addition,Rh2-O action on 3-MA pretreated hepatocellular carcinoma cells resulted in a signifi-cant decrease in extracellular ATP levels(P<0.001).Conclusion:Rh2-O may induce immunogenic cell death by inducing autophagy in hepatocellular carcinoma cells.

Objective:To investigate the effect of ginsenoside octanoate(Rh2-O)on inducing immunogenic cell death in hepa-tocellular carcinoma cells and its molecular mechanism.Methods:Effects of ginsenoside caprylate(Rh2-O)and autophagy inhibitor 3-MA on the activity of hepatocellular carcinoma cells were detected by CCK-8 assay.The effect of Rh2-O on CRT membrane eversion in Hepa1-6 cells were detected by immunofluorescence assay.Rh2-O treated mouse hepatocellular carcinoma cells were used to pre-pare a tumor vaccine for in vivo vaccination experiments in mice.Extracellular ATP levels were detected in real-time.The expression of autophagy-related genes and proteins were measured by real-time fluorescence PCR and Western blot,and the mitochondrial morphol-ogy and co-localization with autophagy proteins were observed by laser confocal microscopy.Results:Rh2-O showed strong cytotoxicity to Hepa1-6 cells[cell viability:(58.54±3.56)%]at a concentration of 150 μmol/L,and a large amount of CRT was observed on the surface of the cell membrane.The tumor emergence rate was 36.36%in the vaccinated group and 100%in the control group.The tumor vaccine prepared by Rh2-O effectively protected mice from the same type of tumor attack;Rh2-O induced an increase in the level of cellular secreted ATP(P<0.05),the mRNA of autophagy-related genes ATG3,p62,LC3 expression levels and autophagy-associated proteins LC3A and LC3B expression levels were increased(P<0.05),and co-localization of mitochondria with autophagy proteins was significantly increased(P<0.05).In addition,Rh2-O action on 3-MA pretreated hepatocellular carcinoma cells resulted in a signifi-cant decrease in extracellular ATP levels(P<0.001).Conclusion:Rh2-O may induce immunogenic cell death by inducing autophagy in hepatocellular carcinoma cells.

Objective:To investigate whether octyl ester derivative of ginsenoside Rh2(Rh2-O)can induce immunogenic cell death of Huh-7 hepatocellular carcinoma cells and possible mechanism.Methods:Huh-7 cells were cultured in vitro,and divided into control group,Rh2-O group,positive control group(mitoxantrone treatment).Viability and apoptosis of cells were detected by CCK-8 and flow cytometry,respectively.Concentrations of high mobility family protein 1(HMGB1)and adenosine triphosphate(ATP)in supernatant were detected by ELISA and chemiluminescence assay,respectively.Membrane eversion of calreticulin(CRT)was detected by immunofluorescence assay.ROS level in cells was detected by fluorescence probe DCFH-DA,and expressions of proteins associated with endoplasmic reticulum stress signaling pathway were detected by Western blot.Results:Rh2-O treatment significantly reduced cell viability,promoted apoptosis,induced secretion of HMGB1,ATP,membrane eversion of CRT,increased accumulation of ROS in cells,and enhanced expressions of endoplasmic reticulum stress-related proteins PERK,eIF2α,p-eIF2α(all P<0.05).After addition of endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA),membrane eversion of CRT induced by Rh2-O was significantly inhibited(P<0.05).Conclusion:Rh2-O can induce immunogenic cell death in hepatocellular carcinoma cells,whose mechanism may be associated with activation of endoplasmic reticulum stress and promotion of CRT membrane eversion.

Objective:To investigate the management of patients with intravenous misplacement of nephrostomy tube following percutaneous renal surgery.Methods:The data of 6 patients with intravenous misplacement of nephrostomy tube during percutaneous nephrolithotomy (PCNL) treated in the two hospitals of Chenzhou from January 2006 to December 2020 were retrospectively analyzed. The median age was 41.0(38.5, 53.0) years old. There were 4 males and 2 females. Three patients had undergone contralateral upper urinary tract operation. One patient had undergone ipsilateral upper urinary tract operation. Two patients had not undergone upper urinary tract operation. Two of the 6 patients had a solitary kidney. Two patients were diagnosed with staghorn calculi (combined with mild hydronephrosis in 1 patient, moderate hydronephrosis in 1 patient). Four patients were diagnosed with ureteral calculus (combined with mild hydronephrosis in 2 patients, moderate hydronephrosis in 1 patient, severe hydronephrosis in 1 patient). In all 6 patients, the tract was dilated with fascial dilators. Immediately after dilator removal, brisk venous bleeding was noted. A nephrostomy tube was inserted promptly through the sheath to tamponade the tract and was immediately closed. Five cases were diagnosed by CT after operation, and 1 case was early diagnosed by intraoperative injection of contrast medium through nephrostomy tube. The nephrostomy tube was misplaced in 5 patients with left upper urinary tract calculi, and in 1 patient with right upper urinary tract calculi. The tip of nephrostomy tube was located in ipsilateral renal vein in 3 patients with left upper urinary tract calculus, inferior vena cava in 2 patients with left upper urinary tract calculus, and contralateral renal vein in 1 patient with right upper urinary tract calculus. No venous thrombosis of renal vein or inferior vena cava was founded in the 6 patients. All 6 patients were managed with strict bed rest, intravenous antibiotics, and one-step or two-step tube withdrawal under close monitoring. One step method referred to total removal of nephrostomy tube under ultrasonic monitoring. Two step method referred to retracting the end of nephrostomy tube into the renal sinus under CT monitoring in the first step, then the nephrostomy tube was completely removed under ultrasound monitoring.Results:All 6 patients were successfully managed with strict bed rest, intravenous antibiotics, and one-step or two-step tube withdrawal under close monitoring. The tube was withdrew by one-step method in 1 patient, by two-step method in 5 patients. The original operations were performed successfully under close observation in 4 patients during the same hospitalization and in 1 patient during the next hospitalization. Other type of operation in 1 patient was performed during the next hospitalization. The all 6 patients were discharged uneventfully. The stone was cleared.Conclusions:Intravenous misplacement of a nephrostomy tube is mainly diagnosed by CT. The nephrostomy tube should be sealed immediately after diagnosis. The intravenously misplaced nephrostomy tube can be successfully removed by one-step or two-step withdrawing under close monitoring. Upper urinary tract stones can be successfully treated at the same time or by stages.

Objective To study the relationship between the red blood cell distribution width-standard deviation（RDW SD） and the risk of recurrent ischemic stroke（IS）. Methods We conducted a retrospective case-control study in 187 patients with recurrent IS who were admitted to neurology department of affiliated hospital of Inner Mongolia medical university，the time of first-ever IS and other baseline characteristic were recorded. The relationship between recurrent IS time and RDW-SD was analyzed by polynomial regression analysis. 187 patients were divides into two groups according to the quartile level of RDW-SD. Kaplan-Meier analysis was used to calculate the cumulative risk of recurrent IS in different groups，and log rank method（χ2 test） was used to compare the differences between groups to study the relationship between RDWSD and recurrent IS risk.Results The results from polynomial regression analysis showed that there was a significant U shaped correlation between RDW-SD level and the recurrent IS time，and its regression coefficient was significantly different from 0 at P<0.05.The median RDW-SD of 187 patients with recurrent IS was 41.80 FL.According to the RDW-SD quartile level，187 patients were divided into 2 groups，including 98 patients corresponding to the highest quartile and the lowest quartile（Q1+Q4）and 89 patients corresponding to the second and third quartiles（Q2+Q3）. There was no significant difference in baseline characteristics between the two groups. Kaplan-Meier analysis illustrated that the patients with the first quartile and the firth quartile（Q1+Q4）of RDW SD levels had a significantly increased cumulative risk of recurrent IS compared to patients with middle quartile（Q2+Q3）of RDW-SD levels（Log Rank=5.047，P=0.02），especially for female patients（Log Rank=6.143，P=0.013）.For male patients，the cumulative risk of recurrent IS was higher in patients with Q1+Q4 group than patients in Q2+Q3 group，but no significant difference was found among two groups（Log Rank=1.325，P=0.25）.Conclusion AU shaped association was found between RDW SD levels and the cumulative risk of recurrent IS，especially for female patients.

Changes in nuclear morphology are common in malignant tumors, but the underlying molecular mechanisms remain poorly understood. Lamins is involved in supporting nuclear structure, and the expression of Lamins is the molecular basis for nuclear morphological changes during tumor progression. In recent years, the research on the relationship between Lamins and malignant tumors has made great progress. Lamins is of great value in the diagnosis, treatment, and prognosis of various malignant tumors.
Cell Nucleus ; Humans ; Lamins/genetics* ; Neoplasms/genetics* ; Prognosis

Objective:To explore the clinical value of peripheral blood lymphocyte subsets in the differential diagnosis of BK virus nephropathy (BKVN) in renal transplantation recipients.Methods:From 2014 to 2018, a total of 172 renal transplant recipients were recruited. Their peripheral blood lymphocyte subsets were detected. According to the pathological puncture results of transplanted kidney, they were divided into acute rejection group (AR, n=68), BKVN group ( n=73) and stable graft function group (STA, n=31). The proportion and absolute number of peripheral blood lymphocyte subsets in each group were measured by flow cytometry and the proportion and absolute count of peripheral blood lymphocyte subsets in each group compared. Results:The proportion and absolute number of CD19 + B cells were markedly lower in BKVN group than those in AR group ( P=0.005, 0.003; 8.5% vs 13.2%, 0.094×10 9/L vs 0.202×10 9/L) and STA group ( P=0.005, 0.003; 8.5% vs 14.8%, 0.094×10 9/L vs 0.198×10 9/L); the proportion of CD3 + CD8 + T cells was significantly higher in BKVN group than that in AR group ( P=0.013; 36.9% vs 31.2%). In addition, no obvious difference existed in the proportion and absolute number of lymphocytes, CD3 + T, CD3 + CD4 + T and CD16 + CD56 + natural killer (NK) among three groups ( P>0.05). No obvious difference existed in the proportion of CD3 + CD4 + / CD3 + CD8 + T cells among three groups ( P>0.05). Conclusions:No difference exists in T cell-related lymphocyte subsets between BKVN and acute rejection recipients. However, the number and proportion of CD19 + B cells decrease markedly in BKVN.

OBJECTIVE:To prepare Idebenone solid dispersion,and to investigate its dissolution rate in vitro. METHODS:Us-ing Poloxamer 407(P407)as carrier,the influence of preparation methods(solvent method,melting method)and the ratio of the drug to P407(1∶1,1∶3,1∶8)on the dissolution of drug were investigated by single factor design. The state of idebenone in ma-trix of solid dispersion was further determined by using differential scanning calorimetry (DSC) and X-ray powder diffraction (XRD). RESULTS:Idebenone solid dispersion prepared by solvent method(the ratio of the drug to poloxamer was 1∶3)showed dissolution rate of 80%. The majority of idebenone existed in the solid dispersion at amorphous forms or molecular state. CONCLU-SIONS:Idebenone solid dispersions with high dissolution rate in vitro is prepared successfully.

Objective: To study the effect of sulforaphane (SUL) on cell growth inhibition, cell cycle, apoptosis and its mechanism in different breast cancer cell lines. Methods: By means of MTT assay, flow cytometry and Western blotting, the effects of the SUL different concentrations on cell growth inhibition, cell cycle arrest, F3Ⅱapoptosis and expression of p34cdc2 and Cdc25C were studied. Results: (1) SUL had strong inhibition effects on the cell growth of tested mammary cancer cell lines, in which the sensitivity of ERP cell lines to SUL was stronger than that of ERN cell line. (2) SUL exhibited obviously G2/M cell cycle arrest to F3Ⅱ and two kinds of ERP cell lines at 5-10?mol/L, whereas no effect on the cell cycle of ERN cell line. (3)The mechanism of hindering transition of F3 Ⅱ cells from G2 phase to M phase was enhancing the phosphorylation of Cdc2, down-regulating the expression of Cdc25C, and inducing inhibition to thedephosphorylation of cyclin B1-Cdc2 complex. (4) No apoptosis was observed under tested conditions. Conclusion: Sulforaphane exhibited obvious differences in the inhibiting effects on the cell proliferation and cell cycle arrests of four different tested breast cancer cell lines, and did not induce apoptosis in F3Ⅱcell line under tested conditions. The mechanism of cell cycle arrest of F3Ⅱcell line appears to involve enhancing the phosphorylation of Cdc2, and down-regulating the expression of Cdc25C.