BACKGROUND: Electrochemotherapy(ECT) is a new and an efficient approach for the treatment of tumor. The technique is very easy to control and operate and is of little harm to normal tissue. Especially, it is very efficient in curing the superficial tumors. In the treatment process of ECT, the selection and constitute of electromagnetic impulse are the key factors.OBJECTIVE: To use the ECT as a therapeutic method for S-180 sarcomas in Kunming mice and obtain the optimal parameters of electromagnetic impulse in curing.SETTING: Department of Wireless Physics, College of Electronics-Information, Sichuan University DESIGN: Random grouping design and controlled experiment study MATERIALS: This experiment was conducted at the Laboratory of Cells, College of Life Sciences, Sichuan University from January 2003 to May 2004. Totally 106 Kunming mice were randomly divided into 12 groups: voltage 500 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups, with 9 mice in each group;voltage 700 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups, with 9 mice in each group; voltage 900 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups,with 9 mice in each group; voltage 700 V, capacitance 10 μF, pulse number 7 and and 9 respectively in 2 groups , with 8 mice in each group , and control group with 9 mice in it.METHODS: The treatment includes injection of Bleomycin 0.04 mg per one (intraperitoneal injecting into tumors through multiple points), followed by application of electromagnetic impulse using the needles. The treatment was taken every three days andthree times in total. Mice were euthanized 18 days later. Tumor volume was measured before irradiation and 10 and 18 days after irradiation. Tumor volume (mm3)=3.141 59×length×width×height/6. Inhibitory rate(%)=(average tumor volume of the control group-average tumor volume after irradiation)/average tumor volume of control group×100%. The relative growth velocity=tumor volume after irradiation/Tumor volume before irradiation.MAIN OUTCOME MEASURES: Tumor volume; inhibitory rate; relative growth velocity RESULTS: Totally 106 animals were enrolled in the experiment and all of them entered the stage of result analysis. ①When the capacitance was 6 μF, the impulse numbers are 5, the voltage altered [for example the 10th day, when voltage was 500, 700, 900 V, the tumor volume was (66.99±91.17),(62.58±71.83),(78.43±73.91) mm3 , respectively]. The effect was the best when the voltage was 700 V, and was the worst when 900 V).②When the voltage and capacitance were fixed and the pulse numbers altered, for example voltage 700 V and capacitance 10 μF, taking the 10th day as the example , the pulse number was 5, 7 and 9, the tumor volume was (80.66±38.17),(41.33±36.40),(39.86±23.03) mm3, respectively. The inhibitory effect was better with the increase of pulse number, and the best when the pulse number was 9.CONCLUSION: According to experimental results, following parameters of electromagnetic impulse, can be concluded: ①The inhibitory effect is the best when the number of impulse is 5, voltage 700 V, capacitance 6 μF. ②The inhibitory effect increases with the increase of pulse number when the voltage is 700 V and capacitance is 10 μF, and is the best when the pulse number is 9.

Objective To investigate the expression of IFN-γ, IL-5, IL-17 and TGF-βby Th cells and Th17 cells in rats with allergic rhinitis upon the intervention of IL-10.Methods SD rats were ran-domly divided into three groups including allergic rhinitis ( AR ) group, IL-10 treated group and control group (n=10).Rats in AR group and IL-10 treated group were sensitized by injection of ovalbumin (OVA) and aluminum hydroxide on the 1st, the 7th and the 14th days.The rats treated with equal volume of saline were set up as the control.The corresponding interventions ( OVA, OVA and IL-10, saline) were respec-tively given to rats in each group on the 21th day for 7 consecutive days.The clinical manifestations in rats were observed within 30 minutes after each administration.Serum samples were collected at 48 hours after the last challenge for the detection of IgE and OVA-sIgE.ELISA and Western blot assay were performed to detect IFN-γ, IL-5, IL-17 and TGF-βin nasal mucosa samples.Results Some characteristic symptoms of AR were observed in rats from AR group and IL-10 treated group.Compared with IL-10 treated rats, rats in AR group showed severe clinical symptoms such as constant rubbing and tearing of the eyes (P<0.05).The levels of IgE and OVA-sIgE in serum samples and the levels of IFN-γ, IL-5, IL-17 and TGF-βin nasal tis-sues were significantly increased in rats with RA (P<0.05), but were reduced with IL-10 intervention (P<0.05).Conclusion Exogenous IL-10 could be used to treat AR by reducing the expression of IFN-γ, IL-5, IL-17 and TGF-βin nasal tissues.

OBJECTIVETo evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.

METHODSA fibroblast cell line (Tri-18; GM02732, 47, XY, +18) was used as the template, and 6 groups were set up in the study. Groups A and B were positive and negative control, respectively; groups C-F were experimental groups involving the MDA products from 1, 2, 5 and 10 cells respectively. In combination of single nucleotide polymorphism (SNP) array, the product of each group was assessed based on the genome coverage, loss of heterozygosity (LOH) rate and allele dropout (ADO) rate.

RESULTSThe nonspecific product of negative control presented an average call rate of 3.2%. The genome coverage of the MDA product increased from 86.4% to 96.4% with the increasing number of template from 1 to 10 cells, while the LOH rate and ADO rate decreased significantly (P<0.05).

CONCLUSIONMDA is a highly efficient and reliable method for whole genome amplification. The fidelity of MDA will be improved significantly with the increasing number of template cells. 10K 2.0 SNP mapping array is a quick, accurate and comprehensive method to evaluate the fidelity of amplified DNA products, but the ADO SNPs should be distinguished from those of preferential amplification from the LOH loci to avoid errors.

Cell Line ; Cells ; cytology ; DNA ; genetics ; Humans ; Loss of Heterozygosity ; Nucleic Acid Amplification Techniques ; methods ; Polymorphism, Single Nucleotide ; Templates, Genetic

Objective To evaluate the value of modified calcofluor white fluorescent staining in the histopathological diagnosis of subcutaneous mycosis,in order to provide a new method for histopathological diagnosis of subcutaneous mycosis.Methods A total of 21 lesional skin tissues were collected from patients with subcutaneous mycosis in the Affiliated Hospital of Chengde Medical University between 1987 and 2017,and embedded in paraffin.Then,each paraffin-embedded tissue section was cut into 4 4-μm-thick serial sections,and subjected to modified calcofluor white fluorescent staining,hematoxylin and eosin (HE) staining,periodic acid Schiff (PAS) staining and Gomori methenamine silver nitrate (GMS) staining respectively.Positive rates and staining outcomes were compared among the above staining methods.Statistical analysis was carried out with SPSS 19.0 software by using chi-square test for comparing the positive rates among the above 4 staining methods.Results Of 21 patients with fungal infections,14 (66.67％) were positive for modified calcofluor white fluorescent staining,5 (23.80％) for HE staining,6 (28.57％) for PAS staining,and 11 (52.38％) for GMS staining.The positive rate by modified calcofluor white fluorescent staining was significantly higher than that by HE staining and PAS staining (x2 =6.718,5.200,respectively,both P ＜ 0.05),while no significant difference was observed between the modified calcofluor white fluorescent staining and GMS staining (x2 =0.693,P =0.530).Conclusion The modified calcofluor white fluorescent staining is an accurate method for detecting fungi,and has a certain application value in the histopathological diagnosis of subcutaneous mycosis.

OBJECTIVE: To explore the pathogenesis for a SRY-negative male with 46,XX disorder of sex development (DSD). METHODS: Peripheral blood samples of the patient and his family members were subjected to chromosomal karyotyping, routine PCR, real-time fluorescence quantitative PCR, whole exome sequencing and whole genome sequencing. The data was analyzed with NextGENe software. RESULTS: Both the proband and his brother presented a 46,XX karyotype with negative SRY gene, while their father presented normal phenotype and karyotype with positive SRY gene. No pathogenic variant associated with sex development was detected by whole exome sequencing, while a 243 kb duplication was detected by whole genome sequencing in the 5' upstream region of the SOX9 gene in the proband, his brother and father. The same duplication was not found in his sister and mother. CONCLUSION The 243 kb duplication at the 5' upstream of the SOX9 gene may predispose to the 46,XX DSD in this family. It is speculated that there exist an unknown core regulatory element in the upstream of the SOX9, and its duplication may trigger expression of SOX9 and initiate testicular differentiation in the absence of SRY gene.
Disorders of Sex Development/genetics* ; Female ; Humans ; Male ; Mutation/genetics* ; Regulatory Sequences, Nucleic Acid/genetics* ; Sex-Determining Region Y Protein/genetics* ; Testis ; Whole Exome Sequencing

Objective: To evaluate the effects of extended nursing mode on the asthmatic children with allergic rhinitis (AR). Methods: Totally116 children aged 6-14 years old with asthma and AR were enrolled to this study from November 2015 to October 2016 in our hospital. They were divided into the regular nursing group and the extended nursing group according to the voluntary principle. The patients in regular nursing group were received routine nursing care in or out of hospital, while the patients in extended nursing group received extended care besides routine nursing. The children were required to record diary about asthma and AR And participate in asthma action projects. Before and after intervention we observed the quantitative score of symptoms and signs, the times of acute attack, the times of oblivion medication, the average days of stay in hospital, the days of fail to School or kindergarten, the lung function and the fractional exhaled nitric oxide (FeNO) in children with asthma and AR within 1 year. These above marks were assessed five times respectively at starting (baseline) , 3 months, 6 months, 9 months and 12 months (1 year) The differences between two groups were compared with appropriate statistical methods. Results: 1 year later, out of 58 cases in extended nursing group, 40 patients (68.97%)were in good control and 18 cases (31.03%) in partial control. Out of 58 cases in regular nursing group, 22 cases (37.93%) were in good control and 36 patients (62.07%) in partial control. There were significant differences between two groups in the effect of disease (χ²=11.23, all P<0.01), the experimental group was significantly better than the control group. The symptoms and signs of allergic rhinitis average scores in regular nursing group were 1.88 ± 0.72, extended nursing group were 0.79 ± 0.71, the difference was statistically significant (t=8.080, P<0.01) with the extended nursing group much better that the regular nursing group. The symptoms and signs of asthma average scores in regular nursing group were1.83 ± 0.75, extended nursing group were 0.88 ± 0.67, the difference was also statistically significant (t=7.133, P<0.01) with the extended nursing group much better that the regular nursing group. In extended nursing groups within 1 year the numbers of acute attack (0.60±0.59), the times of oblivion medication (11.05±7.40), the average days of stay in hospital (8.83±2.79) days,the average days of failing to school or kindergarten (8.69±5.46) days, while in regular nursing group within 1 year the numbers of acute attack(2.94±1.52), the times of oblivion medication (35.28±8.84), the average days of stay in hospital(20.95±5.46 days), the average days of fail to school or kindergarten(24.72±5.92) days, the differences were also statistically significant (t=10.50-15.87, P<0.01). The lung function in extended nursing group (PEF: 82.02±6.04, FEVI: 88.19±5.10, FEV25: 80.67±4.88, FEV50: 80.07±3.73, FEV75: 81.52±3.85) and in regular nursing group (PEF: 79.02±6.12,, FEV1: 80.52±4.72, FEV25: 75.05±7.79, FEV50: 77.59±4.60, FEV75: 78.41±4.19) . The differences were also statistically significant (t=2.90-6.15, P<0.01).The FeNO(13.66±2.87) ppb in extended nursing group and (14.95±3.12) ppb in regular nursing group. There were statistical significant differences between the two groups (t=2.30, P<0.05) with extended nursing group much better than regular nursing group. Conclusion The effect of extended care group is better than that of regular nursing group, and extended care is much more benefit to control asthma and allergic rhinitis in children.

Objective To explore the sen sation recovery of superficial palmar branc h of the radial artery flap with palmar branch of median nerve and donor si te. Methods From January, 2014 to June, 2016, 12 cases of finger soft tissue defects were repaired with sup erficial palmar branch of the radial artery flap with palmar branch of median nerve. The 2 g tactile sensation, 5 g pain sen sation, 30 g pressure sensation and static two-point discrimination (S2 -PD) of the flap was tested regularly. The S2 -PD of the palmar cutaneous branch of the median nerve were recorded respectively on the affected side and the unaffected side. The results were applied comparative t-test to perform statistical analysis, to observe the sensory recovery of the flap and donor site. Results The flaps survived.Twelve cases was followed-up for 6-18 mont hs. The pressure sensation of about 83.3％ of patients recovered after 2 months and 100％ of patients recovered after 3 months. The tactile sensation of about 91.7％ of patients recovered after 3 months and 100％ of patients recovered after 4 months. The pain sensation of about 91.7％ of patients recovered after 4 months of and 100％ of patients recovered after 5 months. S2-PD of the flap was the average of 8.3 mm in 6 months after operation. There were no significant differences in the S2-PD between the affected group [(12.08±2.15)mm] and unaffected group [(10.58±2.11)mm](P>0.05). And the sensory recovery of the control area of the palmar branch of the median nerve was S4 in 2 cases, S3+ in 9 cases, and S3 in 1 case. The sensory recovery was good. Conclusion Superficial palmar branch of the radial artery flap with palmar branch of median nerve can be used repair the skin defect of the fingers. The flap has a good sensery recovery, and the sensation of the donor area is gradually restored.

Objective To explore the influencing factors of the onset of autism spectrum disorder in male children.Methods Totally 151 male children with autism spectrum disorder were selected as case group and 119 healthy male children matched with the age of the case group in the same administrative region were taken as the control group.All children were assessed with the questionnaire for children's autism etiology and risk factors.Results (1) The differences in children having anorexia and partial eclipse (x2 =50.763,P＜0.01),father's age during pregnancy (x2 =11.441,P=0.043),place of pregnancy (x2 =50.763,P＜0.01),hypertension of pregnancy (x2 =5.693,P=0.026),intrauterine hypoxia (x2 =9.332,P=0.002),umbilical cord around the neck(x2 =18.483,P＜0.01),parents smoking and drinking history during pregnancy (x2 =13.660,P=0.008),parental smoking (x2 =12.901,P=0.005) and alcohol consumption (x2 =8.386,P=0.039) during pregnancy,birth height of child (x2 =8.870,P=0.031),amniotic fluid pollution (x2 =4.561,P=0.043),participation time of artificial feeding,major caregivers,delayed development indicators in infants and young children and whether or not the harmonious parent-child relationship were statistically significant(P＜0.05).(2) Children with anorexia and partial diet (OR =12.284,95％ CI =2.768-54.507),living in rural areas during pregnancy (OR =17.251,95％ CI =1.899-1 56.745),parents' history of smoking and drinking (OR =6.191,95％ CI =1.678-22.838),and intrauterine hypoxia during pregnancy (OR=38.859,95％CI=2.944-512.930) may be risk factors for male autism spectrum disorder.Conclusion To correct children's anorexia bias,improve the living environment in pregnancy,reduce pregnancy complications and avoid exposure to tobacco and alcohol pollution during maternal pregnancy can be an effective entry point for the prevention and control of autism spectrum disorders in male children.

BACKGROUND:Tyrosine kinase inhibitors,as well as other types of small-molecule cancer drugs,can cause severe cardiotoxicity. OBJECTIVE:To perform a heart safety re-evaluation by observing the effects of antitumor drugs on isolated heart electrocardiograph,cardiac action potential and associated ion channels and cytotoxicity. METHODS:Extracorporeal cardiac perfusion was given to the isolated rabbit heart using Langendorff perfusion:Sunitinib(0.3,3,10 μmol/L),Crizotinib(0.3,1,3 μmol/L),and Doxorubicin(1,30 μmol/L)were perfused sequentially for 120 minutes to record electrocardiograph and left ventricular pressure.A blank control group was set for comparison.Manual patch clamp was used to record the effects of Crizotinib,Sunitinib,Doxorubicin on hERG,Cav1.2,Nav1.5 channel currents and action potential in human induced pluripotent stem cell derived cardiomyocytes.Adenosine triphosphate level in human induced pluripotent stem cell derived cardiomyocytes was detected by CellTiter-Glo luminescent cell viability assay. RESULTS AND CONCLUSION:Isolated rabbit heart using Langendorff perfusion:Compared with the blank ontrol group,Sunitinib and Crizotinib at≥3 μmol/L decreased heart rate(P<0.01)and prolonged QT/QTc interval(P<0.01),and reduced left ventricular pressure to different extents.Manual patch clamp recording:Compared with the blank control group,Sunitinib and Crizotinib at 3 μmol/L inhibited the activities of hERG,Nav1.5 and Cav1.2 channels and significantly prolonged the duration of action potential(P<0.01).According to the analysis of the test article,the difference between the labeled concentration and the measured concentration of the recovered solution was not significant.Cell viability assays:Compared with the blank control group,adenosine triphosphate content in human induced pluripotent stem cell derived cardiomyocytes significantly decreased after treatment with Sunitinib(IC50=4.64 μmol/L),Doxorubicin(IC50=4.21 μmol/L)and Crizotinib(IC50=2.87 μmol/L),indicating that cell viability significantly decreased(P<0.01).To conclude,this study successfully established an early cardiac safety evaluation method for antitumor drugs,which provides good support and help for the subsequent development of antitumor drugs.

BACKGROUND:Hyperuricemia is a common metabolic disease,and the main clinical manifestation of patients with hyperuricemia is the formation of uric acid crystals leading to gout.Previous studies have only reported that uric acid crystals lead to intervertebral disc degeneration,but there are fewer studies on the correlation between hyperuricemia and intervertebral disc degeneration. OBJECTIVE:To retrospectively analyze the characteristics of intervertebral disc degeneration in patients with hyperuricemia and the correlation between serum uric acid level and intervertebral disc degeneration. METHODS:A retrospective analysis was performed in all patients diagnosed with intervertebral disc degeneration admitted at the Department of Orthopedics,the Affiliated Hospital of Southwest Medical University from January 2021 to December 2022.There were 97 hyperuricemia patients in the hyperuricemia group and 194 non-hyperuricemia patients in the control group according to sex and age in a ratio of 1:2.Blood uric acid test results were collected,and Pfirrmann scoring was performed for the degree of disc degeneration in patients based on the whole spinal MRI images.The difference in the degree of disc degeneration between the two groups was compared,and the correlation between the serum uric acid level and the degree of intervertebral disc degeneration was analyzed. RESULTS AND CONCLUSION:The Pfirrmann score in the hyperuricemia group was higher than that in the control group,and the total number of disc degeneration in the hyperuricemia group was also significantly higher than that in the control group(P<0.05).Spearman correlation analysis showed that the degree of disc degeneration in male patients was positively correlated with serum uric acid level at many spinal segments in the hyperuricemia group(C3/4:r=0.317,C4/5:r=0.333,C5/6:r=0.309,L2/3:r=0.443,P<0.05);the degree of disc degeneration in female patients was also positively correlated with serum uric acid level(C3/4:r=0.354,C4/5:r=0.388,C6/7:r=0.312,T7/8:r=0.282,T9/10:r=0.305,T11/12:r=0.277,L4/5:r=0.319,L5-S1:r=0.367,P<0.05).In the control group,there was no significant correlation between the degree of disc degeneration and serum uric acid level in male and female patients(P>0.05).To conclude,in patients with hyperuricemia,the higher serum uric acid level indicates the more serious intervertebral disc degeneration.Therefore,hyperuricemia is one of the risk factors for intervertebral disc degeneration.