Objective To discuss the activation and biological effects of STAT3 signaling pathway and its negative regulator PIAS3 in glioma cells. Methods Immunohistochemistry(IHC)was used to test the expression of p?STAT3 and PIAS3 in gliomas. AG490(60μmol/L),the in?hibitor of STAT3 signaling pathway,was used to treat U118 and U87 cells for 24/48 h,and the method of MTT assay was taken to evaluate the pro?liferation after AG490 treatment. The expression of p?STAT3 and PIAS3 was also examined by immunofluorescence(IF). Results There was no obvious significance between p?STAT3 and PIAS3 in nuclei or cytoplasm at different grades of gliomas. Whereas,p?STAT3 and PIAS3 were nega?tively correlated in the nuclei of vary grades malignancy gliomas. After AG490 treatment,U118 cells showed no obvious quantitative changes. How?ever,U87 cells showed obvious growth inhibition. IF results showed that there was no significant change at the levels of p?STAT3 and PIAS3 after AG490 treatment in U118 cells. However,the expression of p?STAT3 in the nuclei was down?regulated,and PIAS3 showed obvious nuclear translo?cation in U87 cells. Conclusion Nuclear translocation of PIAS3 plays the key role in modulating JAK/STAT signaling activation and inhibiting glioma cells proliferation.

Based on collecting, arranging, analyzing the literature, the most widely used assessment tools in caring behavior, caring ability and nursing caring characters were introduced. The problems were found out from its application at home and abroad. The aim was to provide the reference that developing and revising the assessment tools of nurses′humanistic that fit for the Chinese local culture.

Objective To evaluate if continuous hemihepatic inflow occlusion(HH)during hepatectomy can be as safe and effective as intermittent total hepatic inflow occlusion(TH)in reducing blood loss during hepatectomy.Methods From November 2001 to March 2006.eighty patients undergoing liver resections were included in a prospective randomized study comparning the intra-and postoperative course underTH(n=40)or HH(n=40).TH was performed with periods of 20 minutes of occlusion and 5 minutes of releasing,while HH with continuous occlusion.The surface area of liver transection was measured and blood loss was calculated.The amount of blood loss,levels of alanine aminotransferuse (ALT)and aspartate aminotransferase(AST),and postoperative course were recorded. Results The total ischemic time of the HH groups was longer than in the TH group[(42±13)min,(31±13)min,P=0.37],and the operative time in the HH group was longer than in the TH group[(236 ±49)min,(204±38)min,P=0.02 ].No signincant difierenee was found between HH and TH group in blood loss during liver parenchyma transection[(500 ±269)ml,(416 ±235)ml,P=0.14]and in the changes of ALT and AST on the first postoperative day[ALT:(677±572)IU/L,(577 ±327)IU/L,P=0.12;AST:(591 ±468)IU/L,(512±301)IU/L,P=0.66].There were no difierences on postoperative morbidity between the two groups(22.5%versus 20.0%,P=0.35).Conclusion The technique of continuous hemihepatic inflow occlusion is as safe and effective as intermittent total hepatic inflow occlusion.

Objective:To evaluate the safety and efficacy of endoscopic cryoablation (ECA) in patients with upper tract urothelial carcinoma (UTUC).Methods:The clinical data of 9 patients with UTUC treated with ECA from April 2018 to September 2019 were retrospectively analyzed. Patients consisted of 3 males and 6 females, with median age of 76 years old (ranging from 50 to 88 years old). Among the patients, 6 cases had tumors of ureter, 1 case had tumor of renal pelvis and 2 cases had tumors of renal pelvis combined with ureter. Of the 9 patients, two had bilateral UTUC, six were presented with single lesion, three were presented with multiple lesion. The size of tumors were (1.53±0.91)cm. The tumors of all cases were localized (≤stage T 2), and there was no carcinoma or suspicious lymph node/distant metastasis. All patients enrolled in this study had strong will to choose kidney-sparing therapy. Biopsy, resection of intraluminal lesion with laser and cryoablation under ureteroscopy or percutaneous nephroscopy was performed under general aneasthesia.Ureteroscopy was performed 3 months after cryoablation. Perioperative complications and follow-up results were recorded and assessed. Results:Cryoablation was successfully performed in patients under ureteroscopy (n=8) or nephroscopy (n=1). The median cryoablation time was 6 (ranging from 4-16) minutes. The median follow-up was 16 months (ranging from 4-24 months). No tumor recurrence was observed at primary sites during follow-up. Two patients with multiple lesions were observed denovo ureteral neoplasms outside the primary sites 3 months and 6 months after cryoablation and treated with second cryoablation. One case died due to cardiovascular events 4 months after surgery. One patient underwent ureteral stricture during follow-up and received ureteroscopic balloon dilatation. No recurrent stricture was found in this case during the subsequent follow-up of 16 months. The other 5 cases showed no recurrence or complications like stricture during follow-up.Conclusions:ECA could probably be a promising treatment for localized UTUC. No recurrence in primary site and low incidence of ureteral stricture was observed during follow-up. The efficacy and safety of ECA need to be verified with large sample study.

Objective To observe changes of hepatic hemodynamics and hepatic functional reserve after splenectomy and periesophagogastric devascularization. Methods From July 2006 to August 2007, thirty patients with portal hypertension caused by hepatitis B cirrhosis underwent splenectomy and periesophagogastric devascuiarization. The PVPG (portal venous pressure gradient) was measured by inductor continually during operation. The HAF (hepatic artery flow), PVF (portal venours flow) and hepatic arterial RI(resistant index) were measured with Doppler sonography. The EHBF(effective hepatic blood flow) and ICGR15 (indocyaninegreen retention rate at 15 minutes) were obtained respectively by indocyaninegreen clearance test before and after the operation. Results PVPG after laparotomy (19±4) mm Hg, ligating the splenic artery(14±4) mm Hg, splenectomy(14±3)mm Hg and periesophagogastric devascularization (12±4) mm Hg showed a tendency to decrease progressively. The PVF decreased [from (42±14) ml/s to (16±8) ml/s] and HAF increased in compensation after operation. The EHBF increased [from (0.48±0.10) L/min to (0.56±0.10) L/min], and the ICGR15 decreased (from 23%±8% to 18%±4%) postoperatively. Conclusion After splenectomy and periesophagogastric devascularization, the hepatic functional reserve improves at least in a short term notwithstanding the decrease of PVPG and PVF.

OBJECTIVE:To establish a method for the content determination of polymer in cefatrizine propylene glycol. METHODS:High performance sephadex gel chromatography was performed on the column of Sephadex G-10 with mobile phase A of 0.01 mol/L phosphate buffer [0.01 mol/L Disodium hydrogen phosphate solution-0.01 mol/L Sodium dihydrogen phosphate solution (61∶39,V/V)](pH7.0)and mobile phase B of water at a flow rate of 1.0 ml/min,the detection wavelength was 254 nm,column temperature was 30℃,and volume injection was 200μl. RESULTS:The linear range of polymer was 2.07-103.30 mg/ml(r=0.999 4);the limit of quantitation of 10.4 ng,limit of detection was 4.1 ng;RSDs of precision and reprodicibility tests were lower than 3%. CONCLUSIONS:The method is specific with high sensitivity and good reproducibility,and can be used for the content determination of polymer in active pharmacentical ingredient cefatrizine propylene glycol.

Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

Objective:To investigate the influence of collateral circulation evaluated by CT perfusion (CTP) imaging in infarction progression and clinical prognoses of patients with acute large-artery occlusion of the anterior circulation before and after thrombectomy.Methods:One hundred and ten patients with acute large-artery occlusion of the anterior circulation within 24 h of onset, admitted to our hospital from May 2018 to September 2019, were chosen in our study; all patients completed thrombectomy; their clinical data were analyzed retrospectively. Regional leptomeningeal collateral-temporally fused maximum intensity projection (rLMC-tMIP) was used to evaluate the collateral circulation based on 4D-CT angiography (4D-CTA). According to the core infarct volume in CTP imaging and the diffusion weighted imaging (DWI) results of MR within one week of surgery, the progressive infarct volume was calculated. Modified Rankin scale (mRS) was used to evaluate the prognoses of these patients 3 months after surgery.Results:(1) There were 56 patients with good collateral circulation and 54 patients with poor collateral circulation. Age ( OR=0.951, 95%CI: 0.910-0.993, P=0.023), cardiac dysfunction ( OR=0.116, 95%CI: 0.018-0.731, P=0.022), baseline fasting blood glucose ( OR=0.788, 95%CI: 0.646-0.961, P=0.019), wakefulness stroke ( OR=0.093, 95%CI: 0.023-0.380, P=0.001), and site of vascular occlusion ( OR=7.604, 95%CI: 2.650-21.821, P=0.000) were independent influencing factors for collateral circulation. (2) Scores of rLMC-tMIP ( 95%CI: -2.947- -1.474, P=0.000), volume of ischemic penumbra ( 95%CI: 0.065-0.126, P=0.000), scores of edema in the brain tissues ( 95%CI: 2.952-7.600, P=0.000), hemorrhage transformation ( 95%CI: 8.966-23.114, P=0.000), and 24 h NIHSS scores ( 95%CI: 0.606-1.248, P=0.000) were independent influencing factors for volume of progressive infarction. (3) There were 59 patients having good prognosis and 51 patients having poor prognosis. Hemorrhage transformation ( OR=0.019, 95%CI: 0.001-0.275, P=0.004) and progressive infarction volume ( OR=0.824, 95%CI: 0.756-0.897, P=0.000) were independent influencing factors for prognoses of patients with acute large-artery occlusion of the anterior circulation after thrombectomy. Conclusion:The scores of rLMC-tMIP based on 4D-CTA can well predict the infarction volume in patients with acute large-artery occlusion of the anterior circulation within 24 h of onset, which can effctively evaluate the clinical prognoses of the patients.

Objective To discuss the effect of PI3K-AKT signaling pathway regulated by microRNA-155(miRNA-155)targeted protein tyrosine phosphatase non-receptor type 21(PTPN21)on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells.Methods Lentivirus transfection was used to silence the expression of miRNA-155 in human Huh7 HCC cells,and real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the silencing effect of miR-155.After obtaining stable cell lines,the cell lines were randomly divided into Blank group(normal Huh7 cells),shNC group(Huh7 cells+empty miR-155 vector),sh-miR-155(Huh7 cells+miR-155 silencing),sh-miR-155+Recilisib group(Huh7 cells+miR-155 silencing+PI3K-AKT agonist),shNC+Recilisib group(Huh7 cells+empty miR-155 vector+PI3K-AKT agonist).Dual luciferase assay was used to determine whether PTPN21 was the downstream of miR-155.The cell proliferation ability of cells in each group was detected by MTT assay.The apoptosis level of each group was tested by flow cytometry.The invasion and migration ability of cells was assessed by Transwell assay.Western blot analysis was used to observe the differences in protein expression of PTPN21,PI3K,P-PI3K,AKT,P-AKT,and apoptosis-related proteins including BAX,BCL-2 and caspase-3 in all groups.Results The expression level of miR-155 in sh-miR-155 group was lower than that in Blank group and shNC group(P＜0.000 1),and the difference in miR-155 expression level between Blank group and shNC group was not statistically significant(P＞0.05).MTT results showed that A values of Huh7 cells at 2,3,4 and 5 day in sh-miR-155 group were lower than those in Blank group and shNC group(P＜0.000 1),while these differences between Blank group and shNC group were not statistically significant(P＞0.05).In sh-miR-155 group the A values at 2,3,4 and 5 day were lower than those in sh-miR-155+Recilisib group and shNC+Recilisib group(P=0.0052 and P＜0.0001,respectively),while the A values at 2,3,4 and 5 day in sh-miR-155+Recilisib were lower than those in shNC+Recilisib group(P＜0.000 1).There was no significant differences in cell migration and number of invasion cells between the Blank group and shNC group(P＞0.05).After activation of PI3K-AKT signaling pathway,the migration and invasion capacity of HCC cells in the shNC+Recilisib group were significantly enhanced when compared with the Blank group(P＜0.000 1).In contrast,the number of migrated and invaded Huh7 cells after miR-155 silencing was significantly lower than that in the Blank group and shNC group(P＜0.000 1)and this phenomenon became reversed by PI3K agonist.Compared with the sh-miR-155 group,in the sh-miR-155+Recilisib group the migration and invasion ability of HCC cells was enhanced(P=0.000 2).Lentiviral transfection of Huh7 human HCC cells to silence miR-155 and downregulate miR-155 inhibiting PTPN21 regulation of the PI3K-AKT signaling pathway,thus inhibiting the invasion,migration and proliferation ability of HCC cells and promoting the apoptosis of HCC cells.Conclusion miR-155 inhibits the migration,invasion and proliferation of HCC cells through targeting PTPN21 regulation of PI3K-AKT signaling pathway.The miR-155 may be a potential therapeutic target for HCC in the future.(J Intervent Radiol,2024,32:44-51)