AIM:To investigate the role of ClC-3 chloride channel in the promotion of radio sensitization of na-sopharyngeal carcinoma CNE-2Z cells by dihydroartemisinin(DHA).METHODS:MTT was used to detect the inhibito-ry effect of DHA on the viability of CNE-2Z cells and normal nasopharyngeal epithelial NP69-SV40T cells,the radio sensi-tization effect of DHA on CNE-2Z cells was detected by cloning assay,the expression of ClC-3 protein was detected by Western blot,the expression of ClC-3 protein was down-regulated by siRNA technology,and the chlorine current of cells was recorded by whole cell patch-clamp technology.RESULTS:(1)Compared with NP69-SV40T cells,DHA selective-ly inhibited the proliferation of CNE-2Z cells,with IC10 values of(13.020±4.831)μmol/L and(5.244±1.050)μmol/L,respectively(P＜0.01).(2)The results of clonal formation experiments showed that DHA had a radio sensitizing effect on CNE-2Z cells,with a radio sensitization ratio of 1.9.(3)DHA could activate the chlorine channel of CNE-2Z cells and produce an outward chlorine current,but had no effect on the chlorine channel of NP69-SV40T cells.(4)DHA promoted the expression of ClC-3 chloric channel protein in CNE-2Z cells(P＜0.01).(5)Chlorine channel blocker NPPB could in-hibit the radio sensitizing effect of DHA on CNE-2Z cells by 1.84 times,and also inhibited the chlorine current activated by DHA.(6)the down-regulation of CNE-2Z ClC-3 protein could inhibit the radio sensitization effect of DHA on CNE-2Z cells by 4.19 times,and the activation of chlorine current by DHA on CNE-2Z cells was no longer produced.CONCLU-SION:DHA has a radio sensitizing effect on nasopharyngeal carcinoma CNE-2Z cells,which is likely to be related to the activation of ClC-3 chloride channel.

Objective:In order to explore the impact of corona virus disease 2019(COVID-19)on the hospitalization of children with bronchiolitis and to improve clinicians′ understanding of the characteristics of bronchiolitis during the COVID-19 epidemic.Methods:This was a multicenter clinical study, and the data have been collected from 23 children′s medical centers in China.All the clinical data were retrospectively collected from children with bronchiolitis who were hospitalized at each study center from January 1, 2019 to December 31, 2021.The results included gender, age at hospitalization, length of stay, respiratory syncytial virus(RSV) test results, severity rating, ICU treatment, and the total number of children hospitalized with respiratory tract infection during the same period.The clinical data of children with bronchiolitis in 2019 before COVID-19 epidemic and in 2020、2021 during COVID-19 epidemic were statistically analyzed and compared.Results:According to a summary of data provided by 23 children′s medical centers, there were 4 909 cases of bronchiolitis in 2019, 2 654 cases in 2020, and 3 500 cases in 2021.Compared with 2019, the number of bronchiolitis cases decreased by 45.94% in 2020 and 28.70% in 2021.In 2019, 2020 and 2021, there were no significant differences in gender ratio, age, and duration of hospitalization.Compared with 2019, the ratio of bronchiolitis to the total number of hospitalizations for respiratory tract infection decreased significantly in 2020 and 2021( χ2=12.762, P<0.05; χ2=84.845, P<0.05).The proportion of moderate to severe bronchiolitis cases in both 2020 and 2021 was lower than that in 2019, and the difference was statistically significant ( χ2=4.054, P<0.05; χ2=8.109, P<0.05).There was no statistically significant difference in the proportion of bronchiolitis cases requiring ICU treatment between 2019, 2020, and 2021 ( χ2=1.914, P>0.05).In 2019, a total of 52.60%(2 582/4 909) of children with bronchiolitis underwent RSV pathogen testing, and among them, there were 708 cases with RSV positive, accounting for 28.00%.In 2020, 54.14%(1 437/2 654) of children with bronchiolitis underwent RSV pathogen testing, and there were 403 cases with RSV positive, accounting for 28.04%.In 2021, 66.80%(2 238/3 500) of children with bronchiolitis underwent RSV pathogen testing, and there were 935 cases with RSV positive, accounting for 41.78%.Compared with 2019 and 2020, the RSV positive rate in 2021 showed a significant increase( χ2=99.673, P<0.05; χ2=71.292, P<0.05). Conclusion:During the COVID-19 epidemic, the implementation of epidemic prevention and control measures reduced the hospitalization rate and severity of bronchiolitis, but did not reduce the positive rate of RSV detection.

Objective:To study the clinical phenotype and molecular genetic characteristics of a Chinese Han family with X-linked retinoschisis (XLRS), and to determine the associated gene variations.Methods:A pedigree investigation was performed.The clinical characteristics and pedigree analysis of a Han Chinese family line with XLRS was conducted in August 2021 at the Xiamen Eye Center Affiliated to Xiamen University.All patients and the carriers underwent comprehensive medical history collection and routine ophthalmological examinations, including visual acuity, non-contact tonometer, slit lamp microscope, direct ophthalmoscope, and optical coherence tomography.The proband and some patients underwent medical optometry, fundus photography or wide-angle fundus photography, and electroretinogram examination.Peripheral venous blood samples were collected from the family members, and whole exome sequencing (WES) analysis was performed on the proband samples.For variants screened by WES, the expanded verification in other patients and normal persons in the family was carried out by Sanger sequencing.Multiple bioinformatic tools were used to analyze the pathogenicity of variants.This study protocol was approved by the Ethics Committee of Xiamen Eye Center of Xiamen University (No.XMYKZX-KY-2021-012). Written informed consent forms were obtained from each subject or guardian of minors.CADD, FATHMM and other bioinformatics tools were used to analyze the pathogenicity of the variation sites.Results:The Han XLRS pedigree consisted of 8 individuals in 3 generations.Out of the 3 cases diagnosed with XLRS based on clinical evaluation, all were male.The mother of the proband was a carrier of related genes.There were 5 persons with normal phenotypes.There was no history of consanguineous marriages within the family, and the disease was shown to be intergenerational, which is consistent with the recessive inheritance of the X chromosome.None of the patients had a history of systemic disease or any other abnormal manifestations.The prevailing feature of ophthalmopathy was poor binocular vision since childhood.The proband and his younger brother had spoke split in the macula, and their grandfather showed atrophy of retinal nerve fibers.Genetic analysis revealed a hemizygous variation c. 214G>C: p.Glu72Gln in the RS1 gene in all the patients in this family.The proband's mother was heterozygous at this site, and all other phenotypically normal family members exhibited wild type at this site.This variant was predicted to be a deleterious variation and likely to cause disease based on bioinformatics analysis. Conclusions:The proband and patients in this Han Chinese family have the known c. 214G>C: p.Glu72Gln hemizygous variation of the RS1 gene and exhibit mild XLRS, which was consistent with the recessive inheritance of X chromosome.

Objective:To investigate the changes of various cytokines and coagulation function in B cell acute lymphoblastic leukemia(ALL) patients with different CRS scores during CD19-CAR-T cell immunotherapy.Methods:87 patients with B-ALL hospitalized in the First Affiliated Hospital of Soochow University and 30 normal controls were enrolled into this study from July 2018 to October 2020. The age of the patients was 32(20, 56) years old and 36(41.4%) were female. All these coagulation indicators, prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer, fibrinogen (Fg) were analyzed by automatic blood coagulation in B-ALL patients before and after treated with CAR-T cell. The ratio of CD19-CAR-T cells and the expression of IL-6, IL-10, IFN-γ, TFN-α, IL-2, IL-4, and IL-17A were analyzed using flow cytometry. The patients′ clinical parameters were detected, and the CRS classification of severity was made according to the standard of consensus.Results:Patients with CRS>3 had prolonged PT and APTT, increased D-dimer, and decreased fibrinogen ( P<0.05). The levels of cytokines of IFN-γ, IL-6, and IL-10 were significantly higher in patients with CRS>3 than that in controls ( P<0.05).The D-dimer level is positively correlated with IL-10. Conclusion:Patients with severe CRS grading have significant coagulation dysfunction in CD19-CAR-T cell immunotherapy. Cytokines IFN-γ, IL-6, and IL-10 may affect coagulation function and CRS grading during CD19-CAR-T cell immunotherapy.

AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.

Objective To analysis the difference of lymph node cleaning and operative complication rate between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer.Methods A summary of 62 patients with thoracic segment esophageal cancer in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from August 2012 to August 2014,who were carried with thoracoscope surgery,were randomly chosed and designed as the thoracosc0Pe group,and 62 patients with thoracic segment esophageal cancer over the same period,carried out with routine thoracotomy,were designed as the control group.All the clinical data of the two groups were collected.The total of thoracic lymph node cleaned and the group of thoracic lymph node cleaned were compared between the two groups.The operation time,intraoperative blood loss,chest tube placement time and postoperative hospital duration were collected and compared.All the patients were followed up at least for one year.The incidence of postoperative complications such as pulmonary infection,pneumothorax,atelectasis,recurrent laryngeal nerve injury and anastomotic leakage in the follow-up period were compared.The follow-up time,mortality and recurrence rate were compared.Results The total of thoracic lymph node cleaned(13.36±3.28) and the group of thoracic lymph node cleaned(3.35±0.84) in the thoracoscope group were lower then these of the control group ((14.22± 2.78) and (3.58±0.75)),but with no statistical difference (t =1.57,1.61,P＞ 0.05).The operation time of the thoracoscope group((314.63±38.72) min) were higher then that of the control((217.46±41.54) min),and the intraoperative blood loss ((205.73 ± 114.38) ml),chest tube placement time ((6.83 ± 1.92) d) and postoperative hospital duration((18.47±5.36) d) of the thoracoscope group were remarkably lower then these of the control ((345.72 ±175.62) m1,(10.04±2.41) d,(22.65±6.84) d,t=13.47,5.26,8.20,3.79,P＜0.05).The incidence of pulmonary infection (4.8％ (3/62)) and atelectasis (1.6％ (1/62)) of the thoracoscope group,were evidently lower then these of the control (17.7％ (11/62),1.3％ (7/62),x2 =5.15,4.81,P＜0.05).There was no significant difference in mortality and recurrence ratebetween the two groups during the follow-up period (3.2％ (2/62) vs.8.1％ (5/62),11.3％ (7/62) vs.14.5％ (9/62),x2 =1.36,0.29,P＞0.05).Conclusion There are no significant difference inlymph node cleaning between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer,but thoracoscope surgery can shorten the length of hospital duration,reduce the intraoperative blood loss,chest tube placement time and postoperative complications.So the thoracoscope surgery is a safe and feasible operation for patients with thoracic segment esophageal cancer.

AIM To explore the radioprotective function of Xihuang Capsules (Bovis Calculus artijactus,Moschus,Olibanum,Myrrha) in patients with acute radiation-induced oral mucositis and its mechanisms.METHODS Eighty patients with nasopharyngeal carcinoma undergone radiotherapy were randomized into two groups:treatment group (radiation plus Xihuang Capsules) and control group (radiation alone).The comparison of two groups in the onset and the tolerated dose of mucositis was made according to the acute radiation injury classification standard.The levels of TNF-α and IL-6 in saliva were detected by ELISA.RESULTS After the treatment with Xihuang Capsules,the onset of oral mucositis delayed significantly and the tolerated dose elevated markedly comparing with the control group (t =2.180,12.930,P ＜ 0.05).The morbidity rate of Ⅲ-N-degree of oral mucositis in the treatment group was lower than that of the control group at the dosage of 40 Gy and 70 Gy,respectively (Z =3.661,4.270,P ＜0.01).Furthermore,there was no difference in the levels of TNF-α and IL-6 in the pre-treatment in the two groups (t =1.010,1.469,P ＞ 0.05).With the increase in radiant dose,the levels of TNF-α and IL-6 in the two groups both elevated dramatically,and peaked at the dose of 40 Gy,but both levels in the treatment group were lower than those of the control group (t =8.305,6.069,P ＜ 0.05).When DT =70 Gy,the levels of TNF-α and IL-6 were lower than pre-radiation,and the levels of TNF-α and IL-6 in the treatment group were lower than those in the radiation alone group (t =3.835,2.488,P ＜ 0.05).CONCLUSION Xihuang Capsules can delay and reduce acute radiation-induced oral mucositis and improve radiation tolerated dose,so it may involve the release of TNF-oα and IL-6 in saliva.

Objective To explore the measurement of(1,3)-β-D glucan in plasma for the diagnosis of pulmonary fungal infections in pulmonary tuberculosis patients. Methods 40 pulmonary tuberculosis patients with pulmonary fungal infections in Guangzhou chest hospital from January 2015 to December 2015 were enrolled as a test group,among which 35 were confirmed and 5 were suspected pulmonary fungal infections. 52 pulmonary tuber-culosis patients without fungal infections were selected as a control group.(1,3)-β-D glucan content(G test)in this 92 patients plasma were detected. The results of G tests were compared with those from etiological diagnosis to assess the performance of G test. Results 13 strains of candida albicans,13 strains of aspergillus,2 strains of candida tropicalis,2 strains of candida glabrata and 6 strains of other yeast were obtained from patients of test group,but no fungal identified from those of control group. The median of G test in test group and in control group was 126.1 and 29.56 pg/mL,respectively,the level in test group was significantly higher than that in control group (P<0.001). 35 cases were identified as positive and 5 were negative in test group by G test ,while 41 cases were identified as negative and 11 were positive in control group. The sensitivity,specificity,positive predictive value, negative predictive value ,concordance and Youden index of G test were 87.5%,78.85%,76.09%,89.13%, 82.6%and 0.663,respectively. Conclusions Candida albicans and aspergillus are more common pathogens than the other fungi isolated from pulmonary tuberculosis patients with pulmonary fungal infection. G test ,used in pul-monary tuberculosis with pulmonary fungal infections diagnosis,is reliable and fast,and has a higher sensitivity, specificity and accuracy.

AIM: To study the effects of disulfiram/copper complex (DSF/Cu) on ultrastructures and mechan-ical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy (AFM) based on the nanoscale resolution and piconewton force measurement level.METHODS: The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10A cells induced by DSF/Cu were compared by flow cytometry.The cell surface morphology, ultra-structure, height, width and roughness were detected by AFM.The effects of DSF/Cu on the hardness (Young’s modu-lus) of the 2 kinds of cells were determined by AFM with indentation technique.RESULTS: DSF/Cu significantly in-duced apoptosis of the MCF-7 cells in a dose-dependent manner, whereas had little effect on the MCF-10A cells.The cell cycle analysis showed that DSF/Cu induced G2 /M arrest in the MCF-7 cells, but led to G0 /G1 arrest in the MCF-10A cells.The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology, and the filopo-dia were retracted obviously, even some became into lamellipodia, or disappeared completely after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.The quantitative analysis indicated that the MCF-7 cells showed smaller width and
larger height, and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.However, little effect in the MCF-10A cells was observed.The biomechanics test at a single cell level demonstrated that the Young’s modulus of the MCF-7 cells and MCF-10A cells were both increased, yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10A cells after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.CONCLUSION: DSF/Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.

[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.