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 investigate the pathogenesis and clinical efficacy of arthroscopic treatment for hallux ganglion cyst deriving from ankle joint.Methods:The clinical data of 21 patients with ankle arthroscopic in the Department of Hand and Foot Surgery,Affiliated Hospital of Jining Medical College from January 2019 to March 2021 were analyzed retrospectively.There were 15 male cases and 6 female cases,aged (52.6±8.2) years (range:42 to 70 years).There were 9 cases of primary operation and 12 cases of recurrence after operation in other hospital.All the patients were examined by ankle arthrography and MRI before operation.The synovial membrane of the ankle was debrided and the tendon sheath of flexor longus was removed at the ankle canal.One year after operation,MRI was performed,and the American Orthopedic Foot and Ankle Society(AOFAS) score of forefoot function and visual analogue scale (VAS) before and after operation were compared by the paired t test or Mann-Whitney U test.The postoperative complications and recurrence were recorded. Results:All patients were operated successfully.The joint capsule at the back of the ankle joint of the patients were ruptured and communicated with the tendon sheath of the flexor longus tendon at the ankle canal.No wound infection,vascular and nerve injury occurred.The follow-up period was (15.0±2.2) months (range:12 to 18 months).During the follow-up period,there was no recurrence of toe appearance and MRI.At the last follow-up,the AOFAS score (90.8±4.3) was significantly higher than that before operation (72.8±6.3) ( t=-10.810, P<0.01),and the VAS score( M(IQR)) was significantly lower than that before operation,the difference was significant (1.0(1.0) vs. 3.0(0.5), Z=-4.081, P<0.01). Conclusions:The possible mechanism of hallux ganglion cyst deriving from ankle joint is that the joint capsule at the back of the ankle joint ruptures and communicates with the tendon sheath of the flexor longus tendon at the ankle canal,and the intra-articular synovial fluid through the cylinder effect generated by sliding with the flexor tendon of the flexor longus tendon in the tendon sheath sac leads to the heel valange cyst.Ankle-synovial cleansing of the ankle joint under ankle arthroscopy and resection of the flexor tendon sheath of the flexor longus tendon at the ankle canal are effective and less invasive.

Objective:To investigate the pathogenesis and clinical efficacy of arthroscopic treatment for hallux ganglion cyst deriving from ankle joint.Methods:The clinical data of 21 patients with ankle arthroscopic in the Department of Hand and Foot Surgery,Affiliated Hospital of Jining Medical College from January 2019 to March 2021 were analyzed retrospectively.There were 15 male cases and 6 female cases,aged (52.6±8.2) years (range:42 to 70 years).There were 9 cases of primary operation and 12 cases of recurrence after operation in other hospital.All the patients were examined by ankle arthrography and MRI before operation.The synovial membrane of the ankle was debrided and the tendon sheath of flexor longus was removed at the ankle canal.One year after operation,MRI was performed,and the American Orthopedic Foot and Ankle Society(AOFAS) score of forefoot function and visual analogue scale (VAS) before and after operation were compared by the paired t test or Mann-Whitney U test.The postoperative complications and recurrence were recorded. Results:All patients were operated successfully.The joint capsule at the back of the ankle joint of the patients were ruptured and communicated with the tendon sheath of the flexor longus tendon at the ankle canal.No wound infection,vascular and nerve injury occurred.The follow-up period was (15.0±2.2) months (range:12 to 18 months).During the follow-up period,there was no recurrence of toe appearance and MRI.At the last follow-up,the AOFAS score (90.8±4.3) was significantly higher than that before operation (72.8±6.3) ( t=-10.810, P<0.01),and the VAS score( M(IQR)) was significantly lower than that before operation,the difference was significant (1.0(1.0) vs. 3.0(0.5), Z=-4.081, P<0.01). Conclusions:The possible mechanism of hallux ganglion cyst deriving from ankle joint is that the joint capsule at the back of the ankle joint ruptures and communicates with the tendon sheath of the flexor longus tendon at the ankle canal,and the intra-articular synovial fluid through the cylinder effect generated by sliding with the flexor tendon of the flexor longus tendon in the tendon sheath sac leads to the heel valange cyst.Ankle-synovial cleansing of the ankle joint under ankle arthroscopy and resection of the flexor tendon sheath of the flexor longus tendon at the ankle canal are effective and less invasive.

Objective:To investigate the effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride (GelMA) hydrogel (hereinafter referred to as composite hydrogel) in the repair of infected full-thickness skin defect wounds in mice.Methods:This study was an experimental study. Cerium oxide nanoenzyme with a particle size of (116±9) nm was prepared by hydrothermal method, and GelMA hydrogel with porous network structure and good gelling performance was also prepared. The 25 μg/mL cerium oxide nanoenzyme which could significantly promote the proliferation of human skin fibroblasts and had high superoxide dismutase activity was screened out. It was added to GelMA hydrogel to prepare composite hydrogel. The percentage of cerium oxide nanoenzyme released from the composite hydrogel was calculated after immersing it in phosphate buffer solution (PBS) for 3 and 7 d. The red blood cell suspension of mice was divided into PBS group, Triton X-100 group, cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group, which were treated with corresponding solution. The hemolysis of red blood cells was detected by microplate reader after 1 h of treatment. The bacterial concentrations of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were determined after being cultured with PBS, cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h. The sample size in all above experiments was 3. Twenty-four 8-week-old male BALB/c mice were taken, and a full-thickness skin defect wound was prepared in the symmetrical position on the back and infected with MRSA. The mice were divided into control group without any drug intervention, and cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group applied with corresponding solution, with 6 mice in each group. The wound healing was observed on 3, 7, and 14 d after injury, and the remaining wound areas on 3 and 7 d after injury were measured (the sample size was 5). The concentration of MRSA in the wound exudation of mice on 3 d after injury was measured (the sample size was 3), and the blood flow perfusion in the wound of mice on 5 d after injury was observed using a laser speckle flow imaging system (the sample size was 6). On 14 d after injury, the wound tissue of mice was collected for hematoxylin-eosin staining to observe the newly formed epithelium and for Masson staining to observe the collagen situation (the sample size was both 3). Results:After immersion for 3 and 7 d, the release percentages of cerium oxide nanoenzyme in the composite hydrogel were about 39% and 75%, respectively. After 1 h of treatment, compared with that in Triton X-100 group, the hemolysis of red blood cells in PBS group, GelMA hydrogel group, cerium oxide nanoenzyme group, and composite hydrogel group was significantly decreased ( P<0.05). Compared with that cultured with PBS, the concentrations of MRSA and Escherichia coli cultured with cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h were significantly decreased ( P<0.05). The wounds of mice in the four groups were gradually healed from 3 to 14 d after injury, and the wounds of mice in composite hydrogel group were all healed on 14 d after injury. On 3 and 7 d after injury, the remaining wound areas of mice in composite hydrogel group were (29±3) and (13±5) mm 2, respectively, which were significantly smaller than (56±12) and (46±10) mm 2 in control group and (51±7) and (38±8) mm 2 in cerium oxide nanoenzyme group (with P values all <0.05), but was similar to (41±5) and (24±9) mm 2 in GelMA hydrogel group (with P values both >0.05). On 3 d after injury, the concentration of MRSA on the wound of mice in composite hydrogel group was significantly lower than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (with P values all <0.05). On 5 d after injury, the volume of blood perfusion in the wound of mice in composite hydrogel group was significantly higher than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively ( P<0.05). On 14 d after injury, the wound of mice in composite hydrogel group basically completed epithelization, and the epithelization was significantly better than that in the other three groups. Compared with that in the other three groups, the content of collagen in the wound of mice in composite hydrogel group was significantly increased, and the arrangement was also more orderly. Conclusions:The composite hydrogel has good biocompatibility and antibacterial effect in vivo and in vitro. It can continuously sustained release cerium oxide nanoenzyme, improve wound blood perfusion in the early stage, and promote wound re-epithelialization and collagen synthesis, therefore promoting the healing of infected full-thickness skin defect wounds in mice.

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 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.