Objective To explore the effect of norcantharidin (NCTD) on proliferation and apoptosis of implanted human gallbladder carcinoma in nude mice. MethodsGBC-SD cells of human gallbladder carcinoma were implanted subcutaneously into nude mice. Mice were randomly divided into control, 5-FU, NCTD and NCTD+5-FU -treatment groups. Tumor size, growth curve and inhibitory rate was respectively evaluated. Cell cycle and apoptosis were measured. Morphological changes of tumorous cells were observed. ResultsLD_ 50 of NCTD for nude mice was 139.96mg?kg -1. Tumor volume (5.61?0.39cm3 vs. 9.78?0.61cm3, P=0.000), percentage of the S phase cells (43.47%?2.83% vs. 69.85%?1.96%, P=0.000) in NCTD group was smaller than that in control group, with tumor inhibitory rate (42.63% vs. 0, P=0.012) and cell apoptosis rate (5.49%?0.59% vs. 15.08%?1.49%, P=0.000) being increased. Compared with other groups,the difference on tumor volume (4.51?1.11 cm3), tumor inhibitory rate (53.89%), percentage of the S phase cells (33.76%?2.39%) and cell apoptosis rate (18.68%?2.38%) in NCTD+5-FU group was statistically significant (P=0.000), with increased nuclear shrinkage, karyorrhexis and typical apoptosis. Conclusion NCTD inhibits the growth of implanted tumor of human gallbladder carcinoma in nude mice. The inhibitory effect could be intensified when combined with 5-FU.

Objective To observe the microstructural changes in lesions of alopecia areata (AA) with dermoscopy and to evaluate their correlation with clinicopathological manifestations. Methods The area of alopecia of 62 patients with AA and 44 patients with other types of hair loss were observed by using a noncontact polarized dermoscope (Dermlite, USA). Clinical data on and laboratory findings from these patients were collected. Pathological examination was carried out with scalp biopsy specimens from the alopecia area of 15 AA patients. Results Characteristic dermoscopic signs of AA included yellow dots, black dots, broken hairs, exclamation mark hairs, short vellus hair and newly-grown short hairs. Among these signs, yellow dots showed the highest prevalence (83.9%). Exclamation mark hairs, black dots and broken hairs were rather specific signs for AA, and the prevalence of the three signs was positively correlated with disease activity and positivity rate of hair-pull test. A positive correlation was also noted between the prevalence of elevated thyroid peroxidase antibody levels and positivity rate of hair-pull test (r = 0.269, P ＜ 0.05 ) as well as prevalence of broken hairs (r = 0.445, P ＜ 0.05), and between the prevalence of yellow dots and that of keratinous plug in follicular orifice. There was a negative correlation between the prevalence of newly-grown short hairs and perifollicular mast cell infiltration and between the prevalence of black dots and the anagen/catagen ratio. Conclusions Yellow dots can serve as a preliminary screening marker for AA. Exclamation mark hairs, black dots and broken hairs are highly sensitive for the confirmation of diagnosis of AA, and often predict progressive AA.Dermoscopic signs are well correlated to the histopathology features of AA, and may be useful for the evaluation of disease severity and guidance on the treatment of AA.

ObjectiveTo assess the relationship of filaggrin expression with atopic diathesis and disease severity in patients with alopecia areata (AA).MethodsThirty-seven patients with AA aged (26.3 ± 10.6) years were enrolled in this study.Atopic diseases were noted in 8 of these patients.Clinical data and laboratory test resuhs were reviewed.Immunohistochemical staining was performed to quantify the expression of filaggrin protein in scalp biopsy specimens from all of the 37 patients with AA and from 10 human controls,and fluorescence-based semiquantitative reverse transcription-PCR to detect the expression of filaggrin mRNA in scalp biopsy specimens from 22 patients with AA and 13 healthy controls.Data were statistically analyzed by Mann Whitney U test,chi-square test,and Spearman's rank correlation test.ResultsThe expressions of filaggrin protein and mRNA were significantly lower in patients with AA than in the controls(P ＜ 0.05 or 0.01 ),and the decrease seemed more obvious in patients with large areas of lesions,long duration of disease,and nail abnormalities,but the degree of decrease was unrelated to the complication with atopic diseases.No significant differences were observed in sex ratio,age at onset,disease duration,area of hair loss,the prevalence of family history or incidence of nail abnormalities and increase in serum IgE and eosinophils,between patients with atopic diseases and those without.ConclusionsThe expressions of filaggrin protein and mRNA are decreased in patients with AA,suggesting that filaggrin may participate in the development of AA and is correlated with the severity of AA.

Objective To detect the expressions of apoptosis-related factors and inflammatory cytokines in superficial and deep layers of as well as anagen hair follicles in lesions of early alopecia areata (AA).Methods Scalp biopsy samples were collected from 25 patients with early AA and 15 healthy human controls.Fluorescence-based quantitative PCR was performed to detect mRNA expressions of apoptosis-related genes p53,caspase 3,Fas,survivin and bcl-2,as well as those of inflammatory cytokines interleukin (IL)-4,IL-10,IL-12 and interferon (IFN)-γ.An immunohistochemical assay was conducted to measure the expression of p53 protein in anagen hair follicles.Results Compared with control skin samples,anagen hair follicles in AA lesions showed significantly increased mRNA expression levels (expressed as 2-△△Ct) of pro-apoptotic factors caspase 3,p53 and Fas (6.78,8.01,9.74,respectively,all P ＜ 0.05),but decreased mRNA expression levels of antiapoptotic factors bcl-2 and survivin (0.08 and 0.03 respectively,both P ＜ 0.01),and similar mRNA expression levels of inflammatory cytokines.There was a significant increase in mRNA expression levels of Th1 cytokines IFN-γ and IL-12 (2.75 vs.1.00,P ＜ 0.05; 85.67 vs.1.00,P ＜ 0.01),but a significant decrease in the expression level of the Th2 cytokine IL-10 (0.002 vs.1.000,P ＜ 0.01) in superficial layers of AA lesions compared with those of normal control skin.The degree of changes in mRNA expression levels of IL-10 and IL-12 was significantly higher in superficial layers than in deep layers of AA lesions (P＜0.01 and 0.05 respectively).The immunohistochemical assay showed that the number of p53-positive cells per 100 cells in anagen hair follicles of AA lesions was higher than that in those of control skin (t =23.79,P ＜ 0.01).Conclusions Anagen hair follicles in AA lesions exhibit high expressions of pro-apoptosis factors,but low expressions of antiapoptotic factors,suggesting that apoptotic factors play a role in the occurrence of AA.

Objective:To explore the diagnosis, treatment and prognostic of pediatric intracranial atypical teratoid/rhabdoid tumor(AT/RT).Methods:A total of 15 pediatric patients with intracranial AT/RT were treated between January 2012 and June 2019 at the First Affiliated Hospital of Zhengzhou University.The clinical data were retrospectively analyzed.Overall survival (OS) rate and progression free survival (PFS) rate were calculated by adopting Kaplan- Meier method.The differences between the 2 groups were tested by performing Log- rank method, and the prognostic factors were analyzed by COX regression. Results:There were 12 males and 3 females, with the median age of 5.5 years (ranging from 8 months to 17.1 years). All patients underwent surgical resection.Gross-total resection (GTR) was achieved in 10 cases and subtotal resection (STR) was carried out in 5 patients.The conducted treatments were as follows: surgery+ radiotherapy+ chemotherapy+ intrathecal injection in 6 cases, surgery+ chemotherapy+ intrathecal injection in 4 cases, surgery+ radiotherapy in 2 cases, and surgery alone in 3 cases.Until January 2020, the median survival time of all the 15 patients was 18 months (ranged 1-27 months), and the survival rate was 33.3%.The 1-year OS rate and PFS rate for all 15 cases were 71.5% and 49.7%, respectively.The 2-year OS rate and PFS rate were 17.9% and 0, respectively. Log- rank analyses revealed that the 1-year OS rates of children less than 3 years old and those older than 3 years were 87.5% and 57.1%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with GTR and those with STR were 90.0% and 40.0%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with tumor dissemination and those without tumor dissemination were 100.0% and 33.3%, respectively( χ2=9.865, P=0.002). The 1-year OS rates of children in the standard-risk group and those in the high-risk group were 88.9% and 41.7%, respectively ( χ2=5.111, P=0.024). COX regression analyses proved that age, the extent of tumor resection, tumor dissemination and risk stratification are independent risk factors for prognosis [hazard radio( HR)=3.411, 3.795, 5.245, 3.397; P=0.025, 0.011, 0.001, 0.017]. Conclusions:Pediatric intracranial AT/RT is rare.The preliminary diagnosis and prognosis are difficult and poor, respectively.The complete resection of tumors with maximal safety remains the primary treatment.Age, the extent of tumor resection, tumor dissemination and risk stratification are independent prognostic factors for AT/RT children.

Objective To explore the features of recurrence of hepatocellular carcinoma (HCC) after liver transplantation (LT) and the related factors.Methods 117 cases of LT for HCC were retrospectively analyzed in a single transplantation center,Medical School of Hanover between Nov 1993 and May 2006.The cumulative survival rates were calculated using the Kaplan-Meier method and the differences of the related factors between various groups were analyzed by Log-rank test.Results 63 cases (58.3％) met the Milan criteria,45 cases (41.7％) exceeded the Milan criteria.Those comply with and exceed The University of California,San Francisco (UCSF) criteria were 73 cases (67.6％) and 35 cases (32.4％),respectively.103 cases were cadaveric donor liver transplantation (CDLT) and 14 cases of living donor liver transplantation (LDLT) with no significant difference between the two groups in postoperative survival rate (P =0.911).Age(P =0.048),microvascular invasion(P =0.001),over UCSF criteria(P =0.013),and the grading of tumor differentiation(P =0.015) were major factors of tumor recurrence after transplantation.Conclusion Appropriate selection criteria,perioperative comprehensive treatment and effective control of recurrence are the keys for improving survival rate of liver transplantation recipients for HCC.

Objective:To explore the clinical efficiency evaluation and prognostic factors of aspiration guided by neuronavigation in the treatment of pediatric brain abscess (PBA).Methods:A total of 47 patients with PBA were treated with aspiration guided by neuronavigation between January 2013 and January 2019 at the First Affiliated Hospital of Zhengzhou University.All clinical data were retrospectively analyzed.According to Glasgow Outcome Scale on discharge, all children were divided into 2 groups, namely good prognosis group and poor prognosis group.Prognostic factors were analyzed by using univariate analysis and binary Logistic regression multivariate analysis. Results:Among the 47 children, 38 children (80.9%) were assigned to the good prognosis group, and 9 children (19.1%) were assigned to the poor prognosis group.Univariate analysis proved that abscess volume>4 cm( χ2=5.650, P=0.017), multiple or multilocular abscess ( χ2=3.258, P=0.027), and abscess located in functional areas ( χ2=6.187, P=0.013) were correlated with poor prognosis.Multivariate analysis revealed that abscess volume>4 cm( OR=5.913, 95% CI: 2.241-25.917, P=0.023) and abscess located in functional areas ( OR=10.519, 95% CI: 3.918-62.513, P<0.001) were independent risk factors for poor prognosis. Conclusion:The treatment of PBA with aspiration guided by neuronavigation is safe, effective and minimal invasive, and the clinical efficiency is satisfactory.Abscess volume>4 cm and abscess located in deepbrain/functional areas are independent risk factors for poor prognosis.

Purpose To investigate the feasibility of CT-based feature tracking technology to quantify left ventricular myocardial strain(MS)and its significance in hypertrophic obstructive cardiomyopathy(HOCM).Materials and Methods A total of 35 HOCM patients who underwent cardiac coronary angiography from March 2019 to December 2021 in the First Affiliated Hospital of the Air Force Military Medical University were retrospectively included,and a total of 60 cases who were negative for cardiac coronary angiography among those who visited the hospital with suspected coronary artery disease were randomly enrolled.Conventional cardiac functional parameters and MS parameters were quantified via post-processing software,and differences of parameters between the groups were analyzed.The diagnostic efficacy of MS parameters for HOCM was further evaluated.Results Compared to the control group,the HOCM group exhibited significant increases in various conventional left ventricular functional parameters,including left ventricular wall thickness,mass,mass index,end-diastolic volume and stroke volume(t=2.119 to 24.861,all P<0.05).However,there were no statistically significant differences in end-systolic volume and cardiac output between the two groups(P>0.05).The global longitudinal and radial strain values of HOCM group were significantly lower than those of control group(t=12.857,-6.427,P<0.01),while the endocardial global circumferential strain of HOCM group was significantly higher than that of control group(t=-2.369,P<0.05).Among MS parameters,global longitudinal strain exhibited the best diagnostic efficacy for HOCM,with an area under the curve of 0.997.A cutoff value of≤20.78%for global longitudinal strain showed that the sensitivity and specificity was 100%and 95%,respectively.Conclusion The MS parameters quantified by the CT-based feature tracking technique are superior to left ventricular ejection fraction in quantifying left ventricular function,with the highest sensitivity and specificity for early myocardial function impairment of longitudinal strain.In addition,the technique has good repeatability and is expected to become a new indicator for the assessment of myocardial function in HOCM.

Objective:To study the amino acid site variation of HIV-1 Tat protein from different parts of AIDS patients with HAD and non HAD and its effect on oxidative stress of U87 cells.Methods:HIV-1 Tat amino acid sequences were analyzed by BLAST and MEGA6 software to study the variation of amino acid sites in four parts of central nervous tissue basal ganglia(BG) and peripheral spleen (SPL) of an HIV-associated dementia (HAD) patient(H) and a non-HAD patient(N), The HIV-1 tat genes were transfected into U87 cell. The green fluorescent protein was observed under microscope to determine the Tat protein expression. The expression of Tat protein in U87 cells was detected by Western blotting. CK-8 method , Western blotting and malondialdehyde (MDA) detection kit were used to study the effect of Tat protein on cell activity, oxidative stress index glutathione peroxidase (GPX), MDA level. Results:Amino acid sequence analysis showed that the key amino acid sites of HIV-1 Tat protein from N-BG, N-SPL, H-BG and H-SPL were different; Tat protein could inhibit the activity of U87 cells, which could be reversed by antioxidant N-acetyl-L-cysteine (NAC). Compared with the control group, the levels of MDA were increased and the expression of GPX protein was decreased in the four experimental groups ( P<0.05). And different sources of Tat protein had different ability to induce oxidative stress, the level of MDA in H-BG group was higher than that in N-BG group( P<0.05). The expression of GPX protein in BG group of both HAD and non-HAD patients was lower than that of SPL group( P<0.05). Conclusions:There are differences in the key amino acid sites of Tat protein in peripheral and central nervous system between HAD and non-HAD patients, and their effects on oxidative stress were also different.

Objective:To investigate the role of HIV-1 negative regulator (Nef) in HIV-1 neuropathogenicity.Methods:Five different HIV-1 nef genes were obtained from the central nervous system (CNS) and peripheral regions (basal ganglia, frontal cortex, meninges, temporal cortex and spleen) of a patient with HIV-1-associated dementia (HAD). The recombinant pcDNA3.1- nef eukaryotic expression vectors were constructed by connecting them with pcDNA3.1 vector and transfected into human glioma cell line U87 respectively. The expression of Nef protein was detected by immunohistochemistry and Western blotting at 22ndhour, 27 th hour, 32nd hour, 37th and 42nd hour after transfection. The result were analyzed quantitatively by JEDA801D and JD-801 software. The supernatant of U87 cells was collected every 5 hours from 27th hour to 62nd hour after transfection. The levels of TNF-α and IL-1 β in the supernatant were determined by ELISA, and the dynamic expression of the two cytokines was analyzed. Results:Five recombinant pcDNA3.1- nef eukaryotic expression vectors of nef genes from different tissues of an HAD patient were successfully constructed and transfected into U87 cells. The result of immunohistochemistry showed that Nef protein began to express at 42nd hour after transfection, which was further confirmed by Western blot. The result of ELISA showed that the levels of cytokines in the supernatant of each group increased gradually with time from 22ed hour to 37th hour after transfection, but there was no significant difference among the groups (TNF-α: F=0.445, P=0.837; F=0.579, P=0.742; F=0.617, P=0.714; F=2.728, P=0.057. IL-1β: F=2.656, P=0.062; F=0.485, P=0.809; F=0.165, P=0.982; F=2.463, P=0.093); The levels of TNF-α and IL-1 β in the experimental group were significantly higher than those in the control group from the 42nd hour ( P<0.05); after 42nd hour, the levels of cytokines in each group gradually decreased, and the levels of TNF-α and IL-1 β remained stable from the 57th hour to the 62nd hour, while the levels of TNF-α and IL-1 β in the experimental group were higher than those in the control group from the 42nd hour to the 62nd hour, the difference was still statistically significant (TNF-α: F=241.310, P<0.001; F=242.638, P<0.001; F=250.114, P<0.001; F=143.877, P<0.001; F=146.172, P<0.001. IL-1β: F=251.578, P<0.001; F=188.816, P<0.001; F=276.240, P<0.001; F=238.136, P<0.001; F=163.361, P<0.001), and there was no significant difference among the experimental groups ( P>0.05). Conclusions:HIV-1 Nef protein can induce and enhance the secretion of TNF-α and IL-1 β in U87 cells. However, the amino acid variation of HIV-1 Nef protein from different sources in an HAD patient had no effect on the secretion of TNF-α and IL-1 β.