Objective To study the inhibitory effects of siRNA targeting survivin on the expression of survivin,as well as the apoptosis,proliferation and invasion of a human melanoma cell line,M14.Methods Two siRNAs targeting survivin were designed,chemically synthesized,and used to construct the recombinant plasmids,pRAT-H1.1/neo-survivin-siRNA1 and pRNAT-H1.1/neo-survivin-siRNA2.Then,recombinant plasmids were transfected into M14 cells mediated by Lipofectamine 2000 reagent.Those cells untransfected or transfected with empty vector served as the control.After culture over various periods of time.cells were collected for the detection of mRNA and protein expression of survivin with RT-PCR and Western blotting,respectively,and for the examination of apoptosis and proliferation of M14 cells by flow cytometry and MTT methods,respectively.Also,Transwell assay was performed to detect the invasive capability of M14 cells.Results A statistical decrease in the mRNA and protein expressions of survivin was observed along with an increase in apoptotic rate(x2=31.55,P＜0.01)in M14 cells transfectcd with siRNA-containing plasmid compared with untransfected and empty vector-transfected cells.As MTT assay indicated,on day 4 after the transfcorion,the proliferation of M14 cells was inhibited by(55.4±4.3)%,(34.5±4.3)%and(13.3±4.6)%,with pRNAT-H1.1/neo-survivin-siRNA1,pRNAT-H1.1/neo-survivin-siRNA2 and empty vector,respectively:there was a significant difference among the three groups(P＜0.05).Decreased invasive capability was noticed in M14 cells transfected with siRNA-containing plasmid compared with untransfected cells(all P＜0.05).Conclusions The plasmid containing siRNA against survivin can specifically inhibit the expression of sarvivin,proliferation and invasion of tumor cells,and induce cell apoptosis.The inhibition of survivin expression by siRNA may be a rational approach to the gene therapy for malignant melanoma.

Purpose: Temozolomide is used in first-line treatment for glioblastoma. However, chemoresistance to temozolomide is common in glioma patients. In addition, mechanisms for the anti-tumor effects of temozolomide are largely unknown. Ferroptosis is a form of programmed cell death triggered by disturbed redox homeostasis, overloaded iron, and increased lipid peroxidation. The present study was performed to elucidate the involvement of ferroptosis in the anti-tumor mechanisms of temozolomide. Materials and Methods: We utilized the CCK8 assay to evaluate cytotoxicity. Levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), iron, and glutathione (GSH) were measured. Flow cytometry and fluorescence microscope were used to detect the production of reactive oxygen species (ROS). Western blotting, RT-PCR and siRNA transfection were used to investigate molecular mechanisms. Results: Temozolomide increased the levels of LDH, MDA, and iron and reduced GSH levels in TG905 cells. Furthermore, we found that ROS levels and DMT1 expression were elevated in TG905 cells treated with temozolomide and were accompanied by a decrease in the expression of glutathione peroxidase 4, indicating an iron-dependent cell death, ferroptosis. Our results also showed that temozolomide-induced ferroptosis is associated with regulation of the Nrf2/HO-1 pathway. Conversely, DMT1 knockdown by siRNA evidently blocked temozolomide-induced ferroptosis in TG905 cells. Conclusion Taken together, our findings indicate that temozolomide may suppress cell growth partly by inducing ferroptosis by targeting DMT1 expression in glioblastoma cells.

Purpose: Temozolomide is used in first-line treatment for glioblastoma. However, chemoresistance to temozolomide is common in glioma patients. In addition, mechanisms for the anti-tumor effects of temozolomide are largely unknown. Ferroptosis is a form of programmed cell death triggered by disturbed redox homeostasis, overloaded iron, and increased lipid peroxidation. The present study was performed to elucidate the involvement of ferroptosis in the anti-tumor mechanisms of temozolomide. Materials and Methods: We utilized the CCK8 assay to evaluate cytotoxicity. Levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), iron, and glutathione (GSH) were measured. Flow cytometry and fluorescence microscope were used to detect the production of reactive oxygen species (ROS). Western blotting, RT-PCR and siRNA transfection were used to investigate molecular mechanisms. Results: Temozolomide increased the levels of LDH, MDA, and iron and reduced GSH levels in TG905 cells. Furthermore, we found that ROS levels and DMT1 expression were elevated in TG905 cells treated with temozolomide and were accompanied by a decrease in the expression of glutathione peroxidase 4, indicating an iron-dependent cell death, ferroptosis. Our results also showed that temozolomide-induced ferroptosis is associated with regulation of the Nrf2/HO-1 pathway. Conversely, DMT1 knockdown by siRNA evidently blocked temozolomide-induced ferroptosis in TG905 cells. Conclusion Taken together, our findings indicate that temozolomide may suppress cell growth partly by inducing ferroptosis by targeting DMT1 expression in glioblastoma cells.

Objective To study the effect of hGC-MSCs from human gastric cancer tissue on cell proliferation, invasion and epithelial-mesenchymal transition in tumor tissue of gastric cancer tumor-bearing mice. Methods BABL/c nude mice were selected as experimental animals and gastric cancer tumor-bearing mice model were established by subcutaneous injection of gastric cancer cells, randomly divided into different intervention groups. hGC-MSCs group were given different amounts of gastric cancer cells for subcutaneous injection, PBS group was given equal volume of PBS for subcutaneous injection. Then tumor tissue volume were determined, tumor-bearing mice were killed and tumor tissues were collected, mRNA expression of proliferation, invasion, EMT-related molecules were determined. Results 4, 8, 12, 16, 20 d after intervention, tumor tissue volume of hGC-MSCs group were significantly higher than those of PBS group and the more the number of hGC-MSCs, the higher the tumor tissue volume; mRNA contents of Ki-67, PCNA, Bcl-2, MMP-2, MMP-7, MMP-9, MMP-14, N-cadherin, vimentin, Snail and Twist in tumor tissue of hGC-MSCs group were higher than those of PBS group, and mRNA contents of Bax, TIMP1, TIMP2 and E-cadherin were lower than those of PBS group. Conclusion hGC-MSCs from human gastric cancer tissue can promote the tumor growth in gastric cancer tumor-bearing mice, and the molecular mechanism includes promoting cell proliferation, invasion and epithelial-mesenchymal transition.

Objective To study the clinicopathologic characteristics of dysplastic nevus. Methods The specimens from 11 patients with dysplastic nevus were studied for their histological characteristics using haematoxylin and eosin staining, and the patients were analyzed for their clinical manifestations. Results Among the 11 patients, 7 had multiple lesions while the remaining 4 had single lesion. Of the 11 studied lesions, 8 had a diameter ≥ 5 mm; 4 had an obscure margin; 6 had an irregular shape; 4 were irregularly pigmented; 6 displayed an erythematous base. Skin biopsy demonstrated that 3 cases were junctional nevus and 8 were compound nevus. Lentiginous proliferation along the dermal-epidermal junction was observed as a typical histological pattern of all cases. The nevus cells proliferated irregularly and tended toward confluence, forming an appearance of “bridging”. Atypical melanocytes spread subepidermally in a pagetoid manner. Extensive proliferation of melanocytes at the epidermal-dermal junction was observed, with some cells extending beyond the dermal nevus component. Cytological criteria for melanocytic atypia included a nucleus, which was polymorphous and larger than that of a keratinocyte, presence of nucleoli, and hyperchromasia as well as variation in nuclear staining. Conclusions It is important to evaluate the relationship between the histopathologic characteristics and clinical phenotypes of dysplastic nevus, which cannot be diagnosed only based on the atypia of its histological appearance.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with genetic epilepsy with febrile seizures plus (GEFS+). METHODS: Clinical data of the proband and his family members were collected. Following extraction of genomic DNA, the proband was subjected to high-throughput sequencing. Candidate variant was verified by Sanger sequencing of the proband and other family members. RESULTS: The pedigree, including 6 patients with febrile seizures from 3 generations, was diagnosed with typical GEFS+. Among them, 2 had febrile seizures (FS), 1 had febrile seizures plus (FS+), and 3 had febrile seizures with focal seizures. High-throughput sequencing revealed that the proband has carried a heterozygous missense variant of c.4522T>A (p.Tyr1508Asn) of the SCN1A gene. Sanger sequencing confirmed that other five patients and one normal member from the pedigree have also carried the same variant, which yielded a penetrance of 85.7%. CONCLUSION The c.4522T>A (p.Tyr1508Asn) of the SCN1A gene probably underlay the disease in this pedigree. The pattern of inheritance was consistent with autosomal dominant inheritance with incomplete penetrance. Above finding has enriched the variant spectrum of the SCN1A gene.
Epilepsy/genetics* ; Humans ; NAV1.1 Voltage-Gated Sodium Channel/genetics* ; Pedigree ; Phenotype ; Seizures, Febrile/genetics*

OBJECTIVETo probe into a method for increasing clinical therapeutic effect on renal hypertension of chronic kidney disease.

METHODSOne hundred and fifty-two cases were randomly divided into a combined acupuncture and medicine group and a medication group, 76 cases in each group. The combined acupuncture and medicine group were treated with acupuncture balance points "Jiangya" and "Shenbing" as main, combined with small dose of hypotensor, Irbesartan; and the medication group were treated with oral administration of Irbesartan and Fosinopril. Their clinical therapeutic effects were compared.

RESULTSAfter treatment of 4 weeks, 56.58% of the patients in the combined acupuncture and medicine group reached to the objective value [DBP < or = 84.96 mm Hg (1 mm Hg= 0.133 kPa)], and 53.95% of the patients in the medication group reached to the objective value, with no significant difference between the two groups (P > 0.05). After treatment of 8 weeks and 24 weeks, the blood pressure-decreasing effect in the combined acupuncture and medicine group was better than that of the medication group (P < 0.01). After treatment, protein in the urine decreased and blood creatinine reduced, the clearance rate of endogenous creatinine raised in the two groups, with very significant differences (P < 0.01).

CONCLUSIONAcupuncture combined with small dose of medicine and simple medicine have a same therapeutic effect on renal hypertension of chronic kidney disease, but with prolongation of treatment time, the therapeutic effect and advantages of the combined acupuncture and medicine therapy were superior to the medication.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Chronic Disease ; Combined Modality Therapy ; Female ; Humans ; Hypertension, Renal ; therapy ; Kidney Diseases ; complications ; Male ; Middle Aged

Objective To analyze the epidemiological,clinical and histopathological characteristics of cutaneous leishmaniasis.Methods This study included six patients with cutaneous leishmaniasis diagnosed in the Institute of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College,over the past 10 years.The epidemiological features as well as clinical and histopathologic presentations of these patients were analyzed retrospectively.Results All of the six patients were male.The mean age at onset of skin eruptions was 47.67 (range:37-67) years,and the mean duration of disease was 10 (range:6-18) months.Clinical presentations included erythema,nodules and ulcers in the face and limbs.Skin biopsy revealed infection-associated granulomatous inflammation with many amastigotes (basophilic bodies) in the cytoplasm of histiocytes,which were highlighted with Giemsa stain.All the patients had a history of working at or travel to epidemic areas.Conclusion The diagnosis of cutaneous leishmaniasis mainly depends on epidemiological data,clinical manifestation and histopathologic findings.

A 24-year-old man presented with a 6-year history of pustules,atrophic scars,and alopecia of the scalp.Dermatological examinations showed generalized alopecia and atrophic scarring of the scalp.A few hairs remained at the vertex region with keratotic papules,pustules,or black sears around the hair follicles.There were also residual hairs at the forehead,temples,and occipital region near the hairline.Clusters of hairs were noted in the occipital region,and keratotic papules were observed around the hair follicles.Erythema,scales,and follicular keratotic papules were found in the superciliary arch.The eyebrows and axillary hairs partially shed and were sparse.Follicular keratotic papules were also found on both cheeks,axillae,chest,abdomen,back and limbs.Stomatological examinations revealed a large space between the upper and lower anterior teeth,anteverted upper anterior teeth,congenital absence of (+1) tooth,deep overbite and deep overjet of anterior teeth.He had a fissured tongue which lacked filiform papillae.Pathological examinations of the skin lesionsshowedathickenedspinouslayer andmassiveinfiltrationsof plasmacytes, neutrophils, and multinucleated giant cells around hair follicles.A diagnosis of folliculitis spinulosa decalvans was made based on the clinical manifestations and histopathological findings.

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.