Objective To study the effects of 5-aza-dc on the expression of IGFBP7 in and proliferation of melanoma cell lines A375 and M14.Methods Reverse transcription-PCR and immunocytochemistry were performed to detect the mRNA and protein expression of IGFBP7 in A375 cells and M14 cells after treatment with 5-aza-dc of 10μmol/L for 48 hours,and MTT assay to measure the proliferation of both cell lines treated with 4 different concentrations (2.5,5,10,20μmol/L) of 5-aza-dc for various durations.Results The treatment with 5-aza-dc restored IGFBP7 expression at both mRNA and protein levels.The four concentrations of 5-aza-dc inhibited the proliferation of A375 and M14 cells in a dose-dependent (F=561.12,271.43,respectively,both P<0.01) and time-dependent (F=141.35,549.33,respectively,both P<0.01) manner.Conclusions DNA methylation may be involved in the modulation of aberrant IGFBP7 gene expression in melanoma,and 5-aza-dc could inhibit the proliferation of A375 and M14 cells.

Objective To improve the understanding of cutaneous intravascular natural killer/T-cell lymphoma (CIVNKTC). Methods Clinical data on five cases of CIVNKTC were collected. The histopathological feature, treatment and prognosis of CIVNKTC were retrospectively analyzed and discussed. Results Of the 5 patients, 1 was male and 4 were female. The age of onset ranged from 38 to 83 years (average, 56.2 years). All the patients presented with multiple plaques and nodules as the primary symptoms. Histopathological examination revealed vasodilatation in the dermis and subcutaneous tissue, as well as atypical lymphoid cells with large hyperchromatic nuclei containing 1-2 small nucleoli in dilated veins. Immunohistochemical studies of tumor cells showed positive staining for CD3ε, cytotoxic proteins (including T cell-restricted intracellular antigen-1, granzyme B and perforin)and Epstein-Barr virus(EBV)-encoded microRNA, but negative staining for cytokeratin, CD20, CD79a, CD4 and CD8. Furthermore, the tumor cells stained positive for CD56 in two patients. Among the 5 patients, only 2 received chemotherapy and the remaining received no treatment. During a 24-month follow-up, 4 patients died, and only 1 survived with the tumor. Conclusion CIVNKTC is a rare extranodal Hodgkin′s lymphoma with distinct histologic manifestations and immunophenotypes, rapid and aggressive clinical course, and poor prognosis.

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.

Objective To study the effects of ankle-foot retractor on ankle joint motion and equilibrium function of stroke hemiplegic patients.Methods 74 patients were divided into two groups: observation group (n=36) and control group (n=38). The control group was trainedby routine rehabilitation training program and electro-uprise bed, the observation group was trained by ankle-foot retractor based on routinerehabilitation training program. The effect was evaluated after 8 weeks. Results After training, either the observation group or the controlgroup showed significant improvement at motion of ankle joint, activity of daily living (ADL), and equilibrium function, but the observationgroup was better than the control group (P<0.001). Conclusion Ankle-foot retractor plays a positive role on motion of ankle joint, ADL andequilibrium function in stroke patients.

The present case report was designed to summarize the clinical experience of operative technique. lung preservation, lung perfusion, and perioperative management. Of 7 cases who underwent allogenic single lung transplantation (LT), 3 were idiopathic pulmonary fibrosis, 2 were chronic obstructive pulmonary disease, 1 was silicosis, emphysema, and bulla, and I was tuberculosis in both sides and presented with destroyed lung in one side. All donors were already brain death. Donor lungs were well preserved utilizing Euro-Colins liquid or low-potassium dextran solution. Donors and recipients were matched in blood type. Of 7 cases selected,5 received single right lung transplantation, and 2 received single left LT. End-to-end anastomosis was performed for pulmonary branches and pulmonary arteries. while atrium-to-atrium anastomosis was performed for pulmonary vein. Antibiotics and immunosuppressants were routinely used prior to and subsequent to LT. Following LT, heart and lung function, usage of antibiotics, and adjustment of immunosuppressant were monitored. Stomal complications regarding bronchus and pulmonary artery and vein did not appear in any patient. Five cases survived for about 2 months, one for approximately 1 year, and one for nearly 2 years. Four cases died of multi-organ failure caused by pulmonary infection, and one of severe pulmonary hemorrhage caused by aspergillus sydowi infection. Rejection occurred in 6 cases. One case sufiered from rejection three times. Selection of indication, selection and preservafton of donor lung, LT operation and pre-and post-operative management of LT have acquired satisfactory achievements. High mortality occurred in patients with preoperative poor cardiac and pulmonary functions and postoperative severe infections accompany with application of immunosuppressant.

BACKGROUND: Lung transplantation can improve quality of life of patients who get terminal pulmonary disease and also it can help to get better survival.Now it has become one of the best therapeutic methods for terminal pulmonary disease.However,limited donors leave the development of lung transplantation in dilemma.The emergence of living lobar transplantation and cadeveric lobar transplantation let this procedure much easier.OBJECTIVE: To evaluate the clinical probability of bilateral lobar transplantation.METHODS: Sequential bilateral lobar transplantation was performed for one 26 years old cystic fibrosis female.Cardiac pulmonary bypass was used during operation.Anti-rejection(Tacrolimus,mycophenolate,etc)and anti-infection was used postoperatively.RESULTS AND CONCLUSION: The recovery course postoperatively was smooth,and the recipient got out of hospital 7weeks later.Bilateral lobar transplantation could offer satisfied short-term pulmonary function.The long term results should be further evaluated.

Objective: Molecular genetics and clinical phenotypic characteristics of 5 alpha reductase deficiency were analyzed. Methods: The genetic results and clinical features classied as Prader grade of external genitalia of 86 children with SRD5A2 mutation seen from 2007 to 2017 at Department of Endocrinology of Beijing Children′s Hospital were analyzed, and the mutation differences in different were compared regions according to the literatures. Results: Among the 86 children, 15 had were homozygous mutations, accounting for 17%, and 71 cases of compound heterozygous mutations accounted for 83%. Totally 172 alleles mutations in this series. The mutation was mainly located on exon 1 and exon 4, in which the mutation frequency of exon 1 was 23.8% (41/172), and the frequency of exon 4 mutation was 55.8% (96/172). A total of 19 mutation types of the SRD5A2 gene in this group were detected, of which 5 were new mutations (p.A228F, p.E57D, p.V124D, p.A117D, p.E197K); 65 patients had p.R227Q mutation, accounting for 76%, while 31 had p.Q6* mutation, accounting for 36%. Other rare types such as p.R246W, p.R103* and so on were also seen in the present study, there was no significant difference between north China and south China (P>0.05). The clinical phenotypes of p.R227Q variation varied, mainly in Prader 3-4, accounting for 82%, while (Prader 0-1) were less, accounting only 2%. The variation of p.Q6* was mainly manifested in Prader 3, accounting for 50%. p.R246Q mainly presented Prader 3. The variation of p.G203S appeared to have Prader 2 and Prader 4-5, accounting for 20% and 73% respectively. There was no significant difference in clinical phenotype corresponding to each protein type (P>0.05) . Conclusion Among the 86 children have identified 19 SRD5A2 mutation types, p.R227Q is a hotspot mutation in Chinese. Variations at different types may have different clinical phenotypes, while the same variations may have different clinical features. There was no significance different in the variation types between the north and the south.

NR5A1 gene mutation is one of the common cause of 46 XY dysplasia (46,XY disorder of sex development,46,XY DSD),which is an autosomal dominant disease.It has wide phenotypes:46,XY gonadal dysplasia is the most common one,site-specific mutations can lead to adrenal dysfunction and may affect the height.In recent years,more and more studies have shown that the mutation of NR5A1 gene can lead to 46,XX ovotesticular DSD and 46,XX testicular DSD.The disease is also characterized by hypergonadotropic hypogonadism,so LH and FSH are high,especially the FSH,leading to a decrease in LH/FSH.The treatment of NR5A1 gene mutation is mainly symptomatic.Gender identification needs to take many factors into consideration.Before puberty,children can use GnRHa to inhibit gonad development and avoid premature ovarian failure.In this review,recent progress of NR5A1 is summarized.

As a digestive organ, the liver has the functions of metabolism, synthesis, and detoxification. It is also an immune organ and plays an important role in maintaining anti-infection, autoimmune stability, and anti-tumor. In particular, the liver has unique immunological advantages. Its immune cells can maintain the liver's immune homeostasis and participate in immunoregulation. A variety of immunotherapy is used in clinical trials for the treatment of difficult and critical liver diseases. This review mainly summarizes the recent clinical trials of immunotherapy in chronic hepatitis B, cirrhosis, hepatocellular carcinoma, and autoimmune liver disease.