Objective To study the effect of siRNA targeting survivin gene on the apoptosis of malignant melanoma cell line A375. Methods The eucaryotic expression vector of pU-survivin-siRNA was constructed and transfected into the A375 cells by electroporation. The protein expression of survivin was examined by Western blotting, and cell apoptosis by flow cytometry. Results The transfection of pU-sur-vivin-siRNA significantly down-regulated survivin expression ( 0.24 ?0.02 in the transfected group versus 0.98 ?0.21 in the control group ) in A375 cells, and promoted cell apoptosis ( 83% in the transfected group versus 28% in the control group, P

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.

Objective Compare the complications between laparoscopic and abdominal cervical cancer surgery, and investigate the safety of laparoscopic cervical cancer surgery and complications classified by Clavien-Dindo classification, then analyze the risk factors. Method Clinical data of 215 cases of cervical cancer received surgery from March 2011 to October 2014 was collected, which include intraoperative, postoperative and postoperative fol﹣low-up data. All the cases were divided into two groups: LRH group (n= 116) and ARH group (n= 88), then ana﹣lyze and compare the difference of intraoperative, postoperative complications and postoperative follow-up data be﹣tween the two groups, assess the safety of the two groups, and statistically concluded related independent risk factors. Results The complications of 204 patients were classified into 4 grades. The criticality and morbidity of intraopera﹣tive and postoperative complications have no significant difference between the two groups (P> 0.05). Logistic re﹣gression analysis show that over criticality grade Ⅱ of LRH intraoperative complication related to operating time, over criticality grade Ⅱ of ARH intraoperative complication related to aged over 50 yr, over criticality grade Ⅱ of ARH intraoperative complication related to BMI> 25. Conclusions Through the analysis of laparoscopic operation and the criticality of complications, the safety of the two groups were no difference. The patients with high risk fac﹣tors should be evaluated comprehensively. And strictly grasp the contraindication and indication.

Objective To evaluate the profile of proliferating T cells and dendritic cells ( DCs ) in the skin infiltrates of patients with mycosis fungoides ( MF) in different stages. Method Paraffin section and immunohistochemisty with monoclonal antibody were used to detect the expression of special antigen per section. Results The numbers of Ki-67+ cells and cutaneous lymphocyte-associated antigen ( CLA+) cells both increased significantly in the skin infiltrates of MF. Most Ki-67+ cells expressed both CLA and CD4 antigen. The number of Ki-67+ cells was significantly higher ( P

An 18-year-old female presented with painful erythema and nodules on both legs for more than 2 years.Dermatological examination showed irregularly sized,mildly indurated,tender,deep subcutaneous nodules arising in diffused infiltrated dark erythematous patches in the inner and posterior region of the left leg.Histopathology showed no significant changes in the epidermis.There were perivascular lymphoid cell infiltrates in the dermis and subcutis.Multiple sites of necrosis of blood vessel walls with vascular occlusions were noted.The lumens of some blood vessels were filled with lymphocytes,among which were many atypical cells with hyperchromatic nuclei and pathologic mitotic figures.Immunohistochemistry showed that lymphocytes in the cavities of blood vessels were positive for CD3(+++),CD3ε(+++),CD2(+),CD56(+++),granzyme B(+++),perforin(+++),CD30(+),Ki67 (+++,100％ ),but negative for CD20,CD5,CD7,CD4,CD8,TdT,anaplastic lymphoma kinase,early membrane antigen (EMA) or pan cytokeratin (pCK).The endothelial cells lining the blood vessels stained positively for CD34.The intravascular lymphocytes were also positive for EBER1/2 by in situ hybridization.A diagnosis of cutaneous intravascular NK/T cell lymphoma was made.

A 20-year-old male patient presented with myalgia of upper limbs and myasthenia of extremities for more than 1 month. Physical examination showed diffuse erythema on the cheeks, upper eyelids, upper chest, neck and dorsa of the hands. The myodynamia of the proximal and distal muscles of upper and lower extremities was grade Ⅳ, Ⅴ, Ⅲ and Ⅴ respectively. Laboratory examinations revealed that the serum levels of creatine kinase, CK-MB and lactate dehydrogenase were 2103 U/L, 83 U/L and 489 U/L respectively, which were all above the normal range. Electromyogram revealed myopathic abnormality and normal nerve conduction velocity. Histopathology of gastrocnemius muscle showed hypertrophy and swelling of muscle fibers, disappearance or fuzziness of transverse striation, and intermuscular lymphoid cell infiltration. A biopsy of the skin lesion from the upper chest showed liquefaction degeneration of and colloid bodies in basal cell layer, perivascular lymphoid cell infiltration in the dermis. A diagnosis of dermatomyositis was established based on the clinical and laboratory findings. After management with intravenous prednisolone 80 mg once daily and symptomatic treatment for 4 weeks, the myodynamia of upper limbs was improved, serum levels of creatine kinase,CK-MB and lactate dehydrogenase reached the normal ranges. However, the myodynamia of lower limbs progressively deteriorated with the emergence of paresthesia. Enhanced MRI scan showed a tumor in the vertebral canal at the level of thoracic vertebra 11 to 12. A spherical encapsulated tumor measuring 3 cm in diameter was surgically removed. The tumor was diagnosed as paraganglioma in vertebral canal according to pathological and immunohistochemical findings. The patient was finally diagnosed with paraganglioma in vertebral canal superimposed on dermatomyositis.

In order to investigate the expression of endothelin receptor B (ETR-B) in human malignant melanoma (MM) cells A375 and SK-mel-1 and the proliferative effects of endothelin 3 (ET3) on A375 cells, RT-PCR was applied to detect the expression of ETR-B gene in human MM cells A375 and SK-mel-1. MTT method was used to evaluate the growth enhancing effects of ET3 on A375 cell line in vitro. The results showed that ETR-B gene was expressed in both MM A375 and SK-mel-1 cells. ET3 had stronger ability to enhance the proliferation of A375 cells in vitro in a concentration-dependent manner. It was suggested that ET3/ETR-B might play an important proliferative role in MM.
Cell Line, Tumor ; Cell Proliferation/*drug effects ; Endothelin-3/*pharmacology ; Melanoma/*metabolism ; Melanoma/*pathology ; Receptor, Endothelin B/*metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective : To investigate whether the RW-splint could be used to guide or determine the CR position of the lower jaw so as to provide help for the later diagnostic design. Methods: 20 class ⅡⅠ malocclusion patients were recruited in orthodontic department of Foshan Stomatological Hospital. They were treated by RW-splint for half a year before orthodontic treatment. The overjet of anterior teeth were recorded before and after treatment. Results : The overjet of anterior teeth was (6.792 ± 0.795) mm before treatment and (7.720 ± 0.930) mm after half a year's treatment. The overjet of anterior teeth had significant difference (t=6.319, P <0.01). The overjet change of anterior teeth between before treatment and half year after treatment was (0.928 ± 0.657) mm. Conclusion The RW-splint wearing before treatment can be used to guide or determine the mandible in the CR position.

The online version of the original article can be found at https://doi.org/10.1631/jzus.B2000190 Erratum to: J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2020 21(10):779-795 https://doi.org/10.1631/jzus.B2000190.

Objective:To understand the pathogen distribution of children with influenza in North China in the past 2018-2019 years, and compare the accuracy of influenza virus antigen test results with that of influenza virus nucleic acid test results, provide reference data for clinical use good influenza virus pathogen detection methods.Methods:Five hundred and eighty throat swab samples of influenza-like children in 10 hospitals, northern China, were collected from December 2018 to January 2019.Each sample was tested by rapid influenza diagnostic test and reverse-transcription polymerase chain reaction(RT-PCR).Results:Of all 580 clinical samples, 256 positive samples (256/580 cases, 44.14%)were detected by the influenza rapid influenza diagnostic test, of which 235 were pure influenza A(235/256 cases, 91.8%), 21 cases were pave influenza B(21/256 cases, 8.2%), and 324 case were negative samples(324/580 cases, 55.86%). No cases were detected positive A and B at the same time.Of all 580 samples were detected using the A /B influenza virus RT-PCR, and a total of 353 cases(353/580 cases, 60.9%) were positive (of which 242 cases were influenza virus antigen-positive), of which 311 were pure A influenza(311/353 cases, 88.1%) and 41 were pure B influenza(41/353 cases, 11.6%), 1 case of mixed infection of A and B(1/353 cases, 0.3%), and 227 cases were negative(227/580 cases, 39.1%). In 324 cases of influenza virus antigen negative samples, 111 cases(111/324 cases, 34.3%) were positive for influenza virus nucleic acid.The detection rate of influenza A in Taiyuan was 23.2% (22/95 cases), and the detection rate of influenza B was 43.2% (41/95 cases), which was significantly different from other regions.With reverse-transcription polymerase chain reaction detection as the standard, the diagnostic value of influenza pathogen detection reagents was evaluated.The sensitivity, specificity, missed diagnosis rate, misdiagnosis rate, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio, Youden index and area under the receiver operating characteristic curve were 68.56%, 93.83%, 31.44%, 6.17%, 94.53%, 65.74%, 11.12, 0.335, 0.624 and 0.812.Conclusions:From December 2018 to January 2019, the majority of children′s influenza in northern China is influenza A virus.Except Taiyuan which is dominated by influenza B. Influenza virus nucleic acid detection has high sensitivity and specificity for diagnosing influenza, and also has the ability to distinguish virus subtypes.Influenza virus antigen detection has a certain diagnostic value, a good specificity (93.83%), sensitivity (68.56%) which needs to be further improved, and a certain rate of missed diagnosis (31.44%) needs to be paid attention to possible missed diagnosis.Detecting positive cases of influenza virus antigens should be given a fast and effective anti-viral treatment, while the negative cases, especially those at high risk for influenza complications, should be confirmed influenza virus RT-PCR as soon as practical.