OBJECTIVETo clone and identify one novel regeneration related gene H(3) (CA854305) from the differential expression genes library we had set up before.

METHODSUse the method of Northern blot to detect the different expressions of the novel gene under different situations, employ the technique of in silico cloning to scan the span of the novel gene, and analyze their sequences. Also we used reverse transcription PCR to validate the largest open reading frame.

RESULTSNorthern blotting results of H(3) (CA854305) showed that the transplanted group had more efficient and extensive expression than untreated and uninjured groups 5 days after spinal cord injury, while the untreated group had more extensive expression than uninjured group. It implied that H(3) might have some relationship with nerve regeneration after spinal cord injury. From the results of in silico cloning we got a longest contig of 1635 bp and an largest open reading frame of 542 bp from 49 to 591 bp correspondent with the Cozak rules. Reverse transcription PCR validated the largest open reading frame sequence primarily.

CONCLUSIONSWe got the sequence of novel gene H(3) which might be one of the regenerationjrelated genes. Key words:Gene library; Genes; Nerve regeneration; Spinal cord injuries.

Animals ; Base Sequence ; Blotting, Northern ; DNA, Complementary ; genetics ; Gene Library ; Molecular Sequence Data ; Nerve Regeneration ; genetics ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Spinal Cord Injuries ; genetics

The proteins encoded by the Herpesviridae β-gene play a critical role in the replication stage of the virus. In this paper, phylogenetic analyses provided evidence that someβ-gene products, such as UL2 and UL23 from HSV1, have their homologous genes in its family, and also exist in prokaryotic organisms, indicating that these viruses appear to have been assembled over evolutionary time by numerous independent events of horizontal gene transfer.

BACKGROUND: There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment. METHODS: We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes. RESULTS: Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100). CONCLUSION 5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.
Animals ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12/genetics* ; Fibroblasts ; Fluorouracil/therapeutic use* ; Humans ; Mice ; RNA, Messenger ; Receptors, CXCR4

The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.

OBJECTIVETo establish transplanted models of VX2 tongue carcinoma in rabbits by three methods and compare these models.

METHODSAfter establishment of VX2 tumor-bearing rabbits, 72 New-Zealand white rabbits were randomly divided into 3 groups. Intact tumour tissue, modified tumour cell suspension, tumour cell suspension were respectively injected into the middle-third lateral border of the tongues of rabbits in 3 groups to induce transplanted VX2 tongue carcinoma. The histological features, the tumour-take rates and the metastasis rates of the 3 models were observed.

RESULTSThe tumour-take rate of 3 models were 83.3%, 91.7% and 33.3% respectively; the lymph node metastasis rates were 71.4%, 100.0% and 37.5% respectively; the lung metastasis rates were 35.7%, 81.3% and 0 respectively. The histological features of the transplanted VX2 tongue carcinoma of 3 models were all consistent with those of moderately differentiated carcinoma.

CONCLUSIONThe biological properties of the transplanted VX2 tongue carcinoma of 3 models is much alike to tongue carcinoma in humans. The model established with modified tumor cell suspension is considered to be more suitable for tongue cancer study.

Animals ; Carcinoma ; Disease Models, Animal ; Humans ; Lymphatic Metastasis ; Neoplasm Transplantation ; Rabbits ; Tongue Neoplasms

Castleman's disease is a slowly progressive and rare lymphoproliferative disorder. Here, we report a 55-year-old woman with superior mediastinal Castleman's disease being misdiagnosed for a long term. We found a 4.3 cm mass localized in the superior mediastinum accompanied with severe clinical symptoms. The patient underwent an exploratory laparotomy, but the mass failed to be totally excised. Pathologic examination revealed a mediastinal mass of Castleman's disease. After radiotherapy of 30 Gy by 15 fractions, the patient no longer presented previous symptoms. At 3 months after radiotherapy of 60 Gy by 30 fractions, Computed tomography of the chest showed significantly smaller mass, indicating partial remission. Upon a 10-month follow-up, the patient was alive and free of symptoms.
Antigens, CD20 ; metabolism ; Castleman Disease ; diagnosis ; immunology ; pathology ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Mediastinal Diseases ; diagnosis ; immunology ; pathology ; radiotherapy ; surgery ; Mediastinum ; diagnostic imaging ; pathology ; Middle Aged ; Multimodal Imaging ; Positron-Emission Tomography ; Radiotherapy, Intensity-Modulated ; Tomography, X-Ray Computed

OBJECTIVETo study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.

METHODSDetermination of the human RhoA sequence as well as the design and constructionof a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into humantongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerasechain reaction was used to examine the mRNA expressionlevels of RhoA. Protein expressions of mRNA, galectin-3,and matrix metalloproteinase (MMP)-9 were evaluated byWestern blot. Transwell invasion assay was performed toassess the invasion ability of tongue carcinoma.

RESULTSRhoA expressions in Tca8113 and SCC-4 cells were reducedsignificantly after transfection of RhoA-siRNA. Protein levels f galectin-3 and MVP-9 were also down-regulated significantly. Invasion ability was inhibited as well.

CONCLUSIONRhoA-siRNA can effectively inhibit RhoA expression in Tca8113 and SCC-4 cells. The invasion ability of tongue carcinoma cells decreased with down-regulation of the protein expressions of galectin-3 and MMP-9, indicating that RhoA-siRNA can inhibit invasion of tongue carcinoma. Results show that RhoA may play an important role in the processes of invasion and metastasis of tongue carcinoma.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Galectin 3 ; metabolism ; Gene Silencing ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection

BACKGROUNDAlthough severe acute respiratory syndrome (SARS) has been controlled, the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method, which would be of great help in clinical diagnosis and also wild host screening. This study aims to establish an effective and rapid serological tool for the diagnosis of SARS-CoV by comparison among whole viral, N and N199 proteins by ELISA.

METHODSSARS-CoV N and N199 (a truncated nucleocapsid gene) genes were cloned, expressed, identified by Western blotting, and applied in screening of human and swine samples. Sera of SARS convalescent-phase patients, normal human sera, sera of patients with other respiratory diseases, and swine sera were screened by ELISA, with whole SARS-CoV F69, N and N199 proteins as antigens.

RESULTSThe sensitivity and specificity of N and N199 proteins in human sera diagnosis were approximate (P = 0.743), which was higher than whole viral protein but the difference was not significant (P = 0.234). The N199 protein proved to be more specific in swine sera screening than whole viral and N protein (P < 0.001).

CONCLUSIONN199 protein is feasible in both clinical diagnosis and SARS-CoV reservoir screening.

Amino Acid Sequence ; Animals ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Molecular Sequence Data ; Nucleocapsid Proteins ; blood ; genetics ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; diagnosis ; Swine

Objective: To analyze the incidence characteristics of occupational diseases in Guangzhou from 2010 to 2020, provide scientific basis for formulating occupational disease prevention and control policies. Methods: In January 2021, based on the data of occupational diseases in Guangzhou reported in the Information Monitoring System of Occupational Diseases and Occupational Health, descriptive epidemiological method was used to analyze the types and characteristics of occupational diseases in Guangzhou from 2010 to 2020. Results: A total of 1341 cases of 38 kinds of occupational diseases in 9 categories were reported in the past 11 years. The incidence of occupational pneumoconiosis, occupational otolaryngology and oral diseases and occupational chemical poisoning ranked the top three, accounting for 38.1% (511/1341) , 30.5% (409/1341) and 16.2% (217/1341) of the total cases respectively. The cases of pneumoconiosis in welders and silicosis accounted for 47.7% (244/511) and 34.4% (176/511) of the cases of occupational pneumoconiosis respectively. The cases of noise deafness accounted for 99.8% (408/409) of occupational otorhinolaryngology oral diseases. Acute occupational chemical poisoning cases accounted for 26.7% (58/217) of the occupational chemical poisoning cases, in which dichloroethane poisoning cases ranked the first, accounting for 79.3% (46/58) . Chronic occupational chemical poisoning cases accounted for 73.3% (159/217) of the occupational poisoning cases, in which benzene and lead poisoning cases ranked the top two, accounting for 79.2% (126/159) and 17.6% (28/159) respectively. Conclusion: Pneumoconiosis, silicosis, noise deafness, benzene poisoning, lead poisoning, dichloroethane poisoning should be supervised and managed as key occupational diseases in Guangzhou.
Benzene ; China/epidemiology* ; Deafness ; Ethylene Dichlorides ; Humans ; Incidence ; Lead Poisoning ; Occupational Diseases/epidemiology* ; Pneumoconiosis/epidemiology* ; Silicosis

ObjectiveTo analyze the expression of Lactate dehydrogenase A（LDHA） in both renal cell carcinoma （RCC） tissue and RCC cell lines， and to investigate the impact of LDHA expression on the progression of RCC. MethodsFrom June 2018 to June 2022， totally 52 cases of RCC tissue samples and 49 cases of para-cancerous tissue samples were collected through surgical procedures from our hospital. LDHA expression was detected using immunohistochemistry （IHC）. The expression levels of LDHA in vitro were also detected in the normal human proximal tubule epithelial cell line HK-2 and renal cell carcinoma cell lines A498， Caki-2， ACHN， and 786-O by using qRT-PCR and Western blot. A recombinant plasmid carrying LDHA-shRNA was constructed and then transfected into 786-O cells to down-regulate the expression of LDHA. Tumor proliferative capacity was monitored using CCK-8 assay， clonal formation assay and EdU assessments. Additionally， cell glycolytic activity was assessed through glucose uptake assay， lactate secretion assay， and ECAR analysis. ResultsIHC analysis revealed significantly higher expression of LDHA in RCC tissue compared to adjacent tissues（P<0.05）. Furthermore， RCC tissues with higher TNM stage exhibited greater expression of LDHA than those with lower TNM stage （P<0.05）. The results of qRT-PCR and Western blot demonstrated that the expression of LDHA in each RCC cell line was significantly higher than that in HK-2（P<0.05）. After blocking the expression of LDHA in 786-O， there was a significant down-regulation of cell proliferation and glycolysis capacity （P<0.05）. ConclusionsThe expression of LDHA in RCC tissue and RCC cell lines is significantly overexpressed compared with normal one， particularly in those with higher TNM stage. Knockdown of the expression of LDHA significantly suppresses cell proliferation and aerobic glycolysis capacity in 786-O.