AIM:To explore the mechanism of PX-12 induced apoptosis of hepatocellular carcinoma cells.METHODS:Human hepatoma cell line Huh7 was se-lected as the main research object.After the cells were treated with thioredoxin inhibitor PX-12,the cell viability was detected by CCK8 method,the cell mi-gration ability was detected by cell scratch test,the cell proliferation ability was detected by cell prolif-eration kit,the levels of intracellular reactive oxygen species and apoptosis were detected by flow cy-tometry,and the expression of apoptosis-related proteins were detected by Western blot.RESULTS:Compared with the control group,the cell viability,migration ability and proliferation ability of PX-12 treatment group were significantly decreased(P＜0.05),and the level of intracellular reactive oxygen species was increased(P＜0.05)in a concentration-dependent manner.Apoptosis inhibitor Z-VAD-FMK and antioxidant NAC could restore the cell viability,and NAC could reduce the accumulation of intracel-lular reactive oxygen species induced by PX-12 and restore the apoptosis induced by PX-12(P＜0.05).CONCLUSION:PX-12 induces apoptosis of hepato-cellular carcinoma cells through oxidative stress.

Objective: To explore the association between circadian rhythm with depressive and anxiety symptoms of college students in Jinzhou City, to provide a theoretical basis for targeted depression and anxiety prevention among college students. Methods: A total of 1 938 college students were selected by convenient sampling method from November to December 2020 for questionnaire survey. The relationship between circadian rhythm and depression and anxiety symptoms was analyzed by using questionnaire,survey including Self Rating Depression Scale (SDS), Self Rating Anxiety Scale (SAS) and Munich Chronotype Questionnaire (MCTQ). Results: There were significant differences in the distribution of depressive symptoms in different majors, smoking, drinking and physical exercise ( χ 2=46.80, 5.88, 5.76, 12.23, P <0.05). There were significant differences in the distribution of anxiety symptoms in different majors, smoking and drinking ( χ 2=9.41, 5.80, 5.56, P <0.05). Stratified analysis showed that the depressive symptoms of different chronotype were statistically varied by age, gender, professional, grade, registered residence, body mass index, smoking, drinking, and sports( χ 2=8.16, 14.42, 12.25, 6.19, 10.99, 15.29, 17.41, 15.63, 7.47, 9.59, 10.51 , P <0.05). The anxiety symptoms of different chronotype were statistically different in age (21 years) and smoking (no), ( χ 2= 8.34 , 7.16, P <0.05). Spearman rank correlation showed that the corrected Mid sleep on Free Days Corrected for Sleep Debt on Work Days (MSFsc) was positively correlated with the standard scores of depression and anxiety ( r s=0.10, 0.09), and social jet lag was positively correlated with the standard scores of depression and anxiety ( r s=0.09, 0.05)( P <0.05). After controlling for age, major, smoking and drinking, binary Logistic regression showed that mean sleep length was inversely correlated with depressive symptoms ( OR =0.82), and weekly insomnia frequency was positively correlated with depressive symptoms ( OR=1.14 ).Early type and intermediate type of chronotypes were negatively correlated with depression ( OR =0.66,0.57). Intermediate type of chronotype was negatively correlated with anxiety symptoms ( OR =0.65). Conclusion Circadian rhythm is related to depressive and anxiety symptoms,among which the average sleep length, early rise and intermediate sleep patterns are negatively related to depression symptoms, and intermediate sleep patterns and anxiety symptoms, suggesting that circadian rhythm disorder may affect depression and anxiety symptoms.

Objective To observe the differences of biological property of glioma stem cells (GSCs) and glioma non-stem cells (nGSCs), and their related protein expressions. Methods The proliferations of GSCs1, GSCs2 and nGSCs1 and nGSCs2 were detected by CCK8 after two, 4, 6, 8, 10 and 12 d of culture in vitro. The sensitivities of the cells to temozolomide (TMZ) were detected by CCK8 after 2 d of culture. The adhesion abilities of cells were tested by adhesion assay. Transwell assay was used to detect the migration and invasion abilities of cells. The activity of matrix metalloproteinase-2 (MMP-2) was detected by gelatin zymography. Western blotting and immunofluorescence staining were used to detect the protein expressions of Notchl and epidermal growth factor receptor (EGFR). Results The survival rate of nGSCs1 was significantly higher than that of GSCs1 and the survival rate of nGSCs2 was significantly higher than that of GSCs2 after 4, 6, 8, 10 and 12 d of culture (P<0.05). The inhibitory concentration (IC)50 of TMZ for GSCs1, nGSCs1, GSCs2 and nGSCs2 was (1536.0±17.67) μmol/L, (514.5±13.44) μmol/L, (2543.0±39.87) μmol/L, (889.6±17.43) μmol/L, respectively (P<0.05). Number of GSCs1 adhering to extracellular matrix proteins Fibronectin and Collagen I was significantly larger than that of nGSCs1, and that of GSCs2 was significantly larger than that of nGSCs2 (P<0.05). The number of migrated GSCs112 and 24 h of cultivation was statistically larger than that of nGSCs1, and that of GSCs2 was statistically larger than that of nGSCs2 (P<0.05). The number of invaded GSCs124 and 36 h of cultivation was larger than that of nGSCs1, and that of invaded GSCs2 was larger than that of nGSCs2, with statistical differences (P<0.05). The activity of MMP2 secreted by GSCs1 was significantly higher than that by nGSCs1, and that of MMP2 secreted by GSCs2 was significantly higher than that by nGSCs2 (P<0.05). Western blotting showed that the relative protein expression level of EGFR/Notch1 in GSCs1 was significantly lower than that in nGSCs1, and that in GSCs2 was significantly lower than that in nGSCs2 (P<0.05). The results of immunofluorescence staining were consistent with those of Western blotting; EGFR protein strongly expressed in nGSCs and weakly expressed in GSCs; Notch1 protein strongly expressed in GSCs and weakly expressed in nGSCs. Conclusion As compared with the high-EGFR-expressing and proliferative primary glioma cells, the high-Notch1-expressing glioma stem cells have higher activity level of MMP-2,stronger abilities of adhesion, migration and invasion, which may be contributed to glioma treatment resistance and its occurrence.

Objective To investigate the expression of FoxO1 gene in idiopathic congenital talipes equinovarus(ICTEV) in Xinjiang Uigur and its correlastion with deformity degree of ICTEV.Methods The FoxO1 protein expression level of foot muscle tissues in 51 Uigur children cases of ICTEV and 25 Uigur normal children were detected by Western blot,the relative expression level of FoxO1 mRNA in different deformity degrees of ICTEV was detected by using real time fluorescence quantitative PCR.Results The Western Blot detection results showed that the expression level of FoxO1 protein in ICTEV children patients was (0.52 ± 0.03),which was lower than(1.61 ± 0.15) in normal children,the difference was statistically significant (P =0.017).The real time fluorescence quantitative PCR detection showed that the relative expression level of FoxO1 mRNA in the Dimeglio type Ⅱ,Ⅲ and Ⅳ were 1.02±0.24,0.67±0.15 and 0.24±0.19 respectively.There was negative correlation between the deformity degree of ICTEV and FoxO1 mRNA relative expression level.Conclusion The transcription level of FoxO1 gene is gradually decreased with the deformity degree aggravation of ICTEV,suggesting that the FoxO1 gene may involve in the whole onset process of Uigur ICTEV.

Chloroplast-based expression system is promising for the hyper-expression of plant-derived recombinant therapeutic proteins and vaccines. To verify the feasibility of obtaining high-level expression of the SARS subunit vaccine and to provide a suitable plant-derived vaccine production platform against the severe acute respiratory syndrome coronavirus (SARS-CoV), a 193-amino acid fragment of SARS CoV spike protein receptor-binding domain (RBD), fused with the peptide vector cholera toxin B subunit (CTB), was expressed in tobacco chloroplasts. Codon-optimized CTB-RBD sequence was integrated into the chloroplast genome and homoplasmy was obtained, as confirmed by PCR and Southern blot analysis. Western blot showed expression of the recombinant fusion protein mostly in soluble monomeric form. Quantification of the recombinant fusion protein CTB-RBD was conducted by ELISA analysis from the transplastomic leaves at different developmental stages, attachment positions and time points in a day and the different expression levels of the CTB-RBD were observed with the highest expression of 10.2% total soluble protein obtained from mature transplastomic leaves. Taken together, our results demonstrate the feasibility of highly expressing SARS subunit vaccine RBD, indicating its potential in subsequent development of a plant-derived recombinant subunit vaccine and reagents production for antibody detection in SARS serological tests.
Chloroplasts ; metabolism ; Cholera Toxin ; Protein Interaction Domains and Motifs ; Recombinant Fusion Proteins ; biosynthesis ; SARS Virus ; Spike Glycoprotein, Coronavirus ; biosynthesis ; Tobacco ; metabolism ; Vaccines, Subunit ; biosynthesis

Objective To research the application of rapid dura-mater reticular incision in the evacuation of acute severe subduml hematoma.Methods The clinical data of 30 patients performed evacuation of acute severe subdural hematoma in our hospital were retrospectively analyzed; the dumra maters was rapidly incised after the first bore hole being drilled,and then the dura mater was incised in reticular shape after the bone flap was formed.The subdural hematoma could effuse slowly and uniformly,and intmcranial hypertension would reduce effectively.Results The subdural hematoma of all patients was removed totally.Based on the Glasgow outcome scale (GOS) scores three months after surgery,8 patients got recovered,13 were referred to moderate maim,5 were severe disability and 4 died.By using this modified method,the acute cenencephalocele rate decreased to 23.3％,which was more effective as compared with that using traditional method (48.2％).Conclusion In acute severe subdural hematoma operation,reticular incision of dura mater may achieve rapid hematoma evacuation and controllable and balanceable decompression,increase curative rate and decrease mortality.

Objective To evaluate the effectiveness of endovascular intervention in treating acute cerebral stroke.Methods Thirty-two patients with acute cerebral stroke (22 with middle cerebral artery occlusion and 10 with internal carotid artery occlusion),admitted to and treated with endovascular intervention in our hospital from January 2010 to October 2013,were chosen in our study;14 were performed arterial thrombolysis+mechanical fragrnengation and 18 were performed mechanical fragrnengation+ solitaireAB stent thrombectomy.Retrospective analysis was performed on the clinical data and treatment efficacy,and the prevention experience of complications was concluded.Results CTA showed that complete recanalization was noted in 18 patients and partial recanalization in 5,with a recanalization rate reaching 7 1％.Postoperative symptomatic intracerebral hemorrhage was noted in 2 patients (6％) and 1 (3.1％) had large area of cerebral infarction.One month after the treatment,MRI indicated decreased areas of infarction in all patients.NIHSS scores were 4-6 points of all patients;favorable prognosis was noted in 23 patients (72％).Nine patients had heavy neurological dysfunction,with limb muscle strength less than grade Ⅲ.Conclusion Endovascular intervention in treating acute cerebral stroke can get high recanalization rate,and achieve good clinical outcome.

Objective To explore the reasons and the treatment counter-measures for secondary intracranial hemorrhage happens in the intracranial tumor craniotomy. Methods Retrospectively analyzed the clinical data of 15 patients with intracranial tumor who suffered secondary intracranial hemorrhage intraoperation. Summarized the tumor characteristics and the situation of corresponding vessels confined by second operation. Results In these 15 cases,the rank of course of disease was 6.5 months to 2 years, mean 1.2 years. The size of the tumor was big with diameter 4.62 ～5. 82cm,mean 5. 12cm,and the tumor was deep surrounding by large range edema,which led to intracranial hypertension. The emissary vein,bridging vein and cortical draining vein were considered as the corresponding vessels for ' secondary intracranial hemorrhage during the second operation carried out for all 15 cases. There wsa no death cases in this research and all patients recovered the nomal ability for self-caring after 3 months following up. Conclusion Sudden drawdown of intracranial pressure and perfusion pressure breakthrough of local vessels had relationship with secondary intracranial hemorrhage during craniotomy for intracranial tumor. Accurate judgement for the occurrence of secondary intracranial hemorrhage intra-operation and quickly taking the effective corresponding measures was the important strategy for prognosis improving for these patients.

Objective To discuss the diagnosis and treatment of superior sagittal sinus thrombosis.Methods The early diagnosis and treatment of superior sagittal sinus thrombosis was induced and summarizd,7 cases in our hospital treated from 2005.6 to 2008.9.were analyzed.Results In this investigation,3 cases were puerperal women,and the other 4 had no special medical history.Brain edema of difference degree was found in all cases on CT scans,including sulcus (7 cases),compression of cistern(5 cases) or even disappeared of cistern(2 cases).CT scanning also revealed dense triangle in 2 cases,multiple small encephalomalacia focus of frontal lobe lesions in 3 cases,and sub-arachnoid hemorrhage(SAH) in 2 cases.The digital subtraction angio-grapby(DSA) also was underwent,of 2 superior sagittal sinus not seen,of 4 appear unevenness,of 7 Arterio-venous circulation delayed and of 5 collateral circulation vasodilatation.After treatment,the clinical symptoms and signs of all patients were improved.All of them could be self-supporting.Followed up for 3 years,the mortality was 0,and no one cerebral hemorrhage again.Conclusion The early diagnosis and treatment of superior sagittal sinus thrombosis could reduce the morbidity and mortality, also could improve prognosis.Especially for the one without cerebral hemorrhage,using heparin at the beginning was an effective and safe way.

Objective To explore hematoma piercing attract joint hematoma clearance to bone valves decompression technique in hematoma volume,skull,pressure high blood pressure brain hemorrhage treatment,and to further explore hypertension brain hemorrhage of reasonable surgery. Methods Hematoma in relatively large quantities of high intracranial pressure in hypertensive cerebral hemorrhage patients were randomly divided into experimental group and control group,and patients in the experimental group were implemented early hematoma puncture to attract a joint hematoma decompressive craniectomy treatment,and the control group were implemented the traditional hematoma de-compressive craniectomy treatment. The prognosis of the two groups were compared. Results The two groups were followed up for 6 months,and evaluated by ADL grade,between the two groups was statistically significant difference in ADL classification( P<0.01). Conclusion In the hematoma volume larger, high intracranial pressure in hypertensive cerebral hemorrhage in the surgical treatment of patients,the early line to attract a joint hematoma puncture decompressive craniectomy was a reasonable and feasible surgical method which can improve these patients prognosis.