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1.
Zhonghua Wai Ke Za Zhi ; 61(9): 801-806, 2023 Sep 01.
Artigo em Chinês | MEDLINE | ID: mdl-37491174

RESUMO

Objective: To examine the feasibility of the modified gasless trans-subclavian approach endoscopic thyroidectomy for lateral neck dissection (LND) in papillary thyroid carcinoma (PTC). Methods: The clinical data of 31 patients with PTC who underwent modified gasless trans-subclavian approach endoscopic LND in the Department of Head and Neck Surgery, Run Run Shaw Hospital, from January to October 2022 were retrospectively analyzed. There were 2 males and 29 females, aged (32.6±8.3) years (range: 17 to 55 years). The maximum diameter of the primary thyroid lesion (M(IQR)) was 1.06 (1.16) cm (range: 0.53 to 2.44 cm), and the maximum diameter of the metastatic lymph node was (1.04±0.37) cm (range: 0.44 to 1.88 cm). Operation time, postoperative hospital stay, number of lymph nodes dissected, and postoperative complications were recorded. Outpatient follow-up was conducted until November 30, 2022. Results: All operations were successfully completed with the endoscopy approach without conversion to open surgery. The operation time was 160 (20) minutes (range: 100 to 215 minutes), and the postoperative hospital stay was 4 (2) days (range: 2 to 14 days). The number of lymph nodes obtained by dissection in the central and lateral compartment of the neck was 11 (12) (range: 0 to 37) and 34.7±14.8 (range: 15 to 69), respectively. Temporary hypoparathyroidism occurred in 4 cases and all recovered within 1 month after the operation. One case suffered from recurrent laryngeal nerve injury (continuing followed up to assess whether it is a temporary injury). The complication of LND included 1 case of chylous leakage that was recovered with conservative treatment, 1 case of Horner syndrome returned to normal 3 months after surgery. During follow-up, there was no residual tumor or recurrence. Conclusion: The modified gasless trans-subclavian approach endoscopic LND for PTC is feasible, with a thorough dissection and concealed incision.


Assuntos
Carcinoma Papilar , Neoplasias da Glândula Tireoide , Masculino , Feminino , Humanos , Câncer Papilífero da Tireoide/cirurgia , Esvaziamento Cervical , Neoplasias da Glândula Tireoide/cirurgia , Neoplasias da Glândula Tireoide/patologia , Estudos Retrospectivos , Carcinoma Papilar/cirurgia , Endoscopia , Tireoidectomia
2.
Zhonghua Wai Ke Za Zhi ; 60(2): 154-158, 2022 Feb 01.
Artigo em Chinês | MEDLINE | ID: mdl-35012275

RESUMO

Objective: To examine the safety and feasibility of gasless submental-transoral combined appoach endoscopic thyroidectomy for papillary thyroid carcinoma (PTC). Methods: A retrospective analysis of the clinical data of 41 patients with PTC who underwent the gasless submental-transoral combined appoach endoscopic thyroidectomy at the Department of Head and Neck Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine from November 2020 to April 2021. There were 5 males and 36 females with the age of (35.0±8.7) years (range: 19 to 58 years). A horizontal incision with a length of 2.0 cm is made under the chin as an observation hole, a 10 mm Trocar and a self-developed retractor are inserted, and two 5 mm longitudinal incisions are made on the labial side in the vestibule of the oral cavity as an operation hole, each inserting a 5 mm Trocar, the operation direction is from the cranial side to the caudal side. The sensation of the lower lip and chin was measured on the first day and one month postoperative. The operation time, hospital stay, the number of lymph nodes dissected and postoperative complications were recorded. Results: Surgical procedures in all cases were successfully completed under endoscopic approach without transfering to open surgery. The operation time was (99±34) minutes (range: 50 to 180 minutes) and the postoperative hospital stay was (3.4±2.2) days (range: 2 to 16 days). The maximum diameter of PTC was (7.6±5.8) mm (range: 2 to 30 mm), and the number of lymph nodes of the central compartment dissection was 6(5) (M(IQR)) (range: 1 to 25). The duration of follow-up is 1 month after operation, and the follow-up method is adopted in outpatient clinic. Postoperation complications included 2 cases of transient hypoparathyroidism, One case of recurrent laryngeal nerve injury (continue to follow up to assess whether it is a temporary injury). Postoperative minor chyle leak, seroma, and local redness and swelling in 1 case each were cured after conservative treatment. 1 case of transient minor numbness of the lower lip was observed. No permanent hypoparathyroidism, postoperative bleeding and numbness of the chin was observed. Conclusion: The gasless submental-transoral combined appoach endoscopic thyroidectomy is a feasible approach in selected PTC patients and has clinical application value.


Assuntos
Neoplasias da Glândula Tireoide , Tireoidectomia , Adulto , Endoscopia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esvaziamento Cervical , Estudos Retrospectivos , Câncer Papilífero da Tireoide/cirurgia , Neoplasias da Glândula Tireoide/cirurgia , Adulto Jovem
3.
Hum Reprod ; 32(4): 836-852, 2017 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-28184438

RESUMO

Study question: What is the chance of clinical pregnancy when fallopian tube catheterization is used for proximal tubal obstruction? Summary answer: The pooled clinical pregnancy rate of tubal catheterization after proximal tubal obstruction is 27% (95% CI 25-30%). What is known already: Restoring fallopian tube patency by performing tubal catheterization has fallen out of favour since the increased availability of IVF. Our study is the first systematic review and meta-analysis to investigate reproductive outcomes following tubal catheterization for proximal tubal obstruction. Study design, size, duration: We undertook a systematic review and meta-analysis of 27 observational studies consisting of 1720 patients undergoing tubal catheterization for proximal tubal obstruction, who attempted to conceive naturally after the procedure. Participants/materials, setting, methods: Systematic literature searches were performed in MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials. A total of 2195 titles and abstracts were reviewed. Only studies that reported outcomes when tubal catheterization was performed with no other tubal surgery were included. Twenty-seven cohort studies matched the inclusion criteria for the meta-analysis. Main results and the role of chance: The meta-analysis showed a pooled clinical pregnancy rate of 27% (95% CI 25-30%) after the use of tubal catheterization for unilateral or bilateral proximal tubal obstruction (27 studies, 1556 patients). In women with bilateral obstruction (14 studies, 617 patients), the clinical pregnancy rate was 27% (95% CI 23-32%). Our meta-analysis demonstrated that the pooled cumulative clinical pregnancy rates were 22.3% (95% CI 17.8-27.8%) at 6 months, 25.8% (95% CI 21.1-31.5%) at 9 months, 26.4% (95% CI 23.0-30.2%) at 12 months, 26.0% (95% CI 22.8-29.7%) at 18 months, 27.0% (95% CI 24.0-30.5%) at 24 months, 27.9% (95% CI 24.9-31.3%) at 36 months and 28.5% (95% CI 25.5-31.8%) at 48 months. The pooled live birth rate (14 studies, 551 patients) was 22% (95% CI 18-26%). The pooled ectopic pregnancy rate (27 studies, 1556 patients) was 4% (95% CI 3-5%). The included studies scored satisfactorily on the Newcastle-Ottawa quality assessment scale. Limitations, reasons for caution: The pooled clinical pregnancy rate after tubal catheterization was found to be almost comparable to that after IVF. However, included studies were small, non-comparative series with significant clinical heterogeneity in population characteristics, follow-up and surgical equipment, technique and experience. Wider implications of the findings: These findings suggest fallopian tube catheterization as an alternative strategy to IVF in patients presenting with proximal tubal obstruction. Further research should focus on comparing different surgical techniques of fallopian tube catheterization with IVF and provide cumulative reproductive outcomes over long-term follow-up. Study funding/competing interest(s): No funding was required and the authors have no competing interests to declare. Registration number: N/A.


Assuntos
Doenças das Tubas Uterinas/cirurgia , Infertilidade Feminina/cirurgia , Taxa de Gravidez , Adulto , Cateterismo/métodos , Feminino , Humanos , Gravidez , Resultado do Tratamento
4.
Scand J Rheumatol ; 36(4): 278-84, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17763205

RESUMO

OBJECTIVES: To investigate the association between tumour necrosis factor-alpha (TNFalpha), TNF receptor superfamily member 1A (TNFRSF1A, also known as TNFRI), TNFRSF1B (TNFRII), and interleukin-1beta (IL-1beta) single nucleotide polymorphisms (SNPs) and the susceptibility to persistent palindromic rheumatism (PR). METHODS: Fifty-six unrelated patients with persistent PR and 100 unrelated healthy controls were genotyped for TNFalpha -308G/A, -238G/A, and +488G/A, TNFRSF1A -609G/T and +36A/G, TNFRSF1B +676T/G and +1663G/A, and IL-1beta -511C/T, -31T/C, and +3954C/T using real-time polymerase chain reaction (RT-PCR). RESULTS: The TNFRSF1A +36G allele [odds ratio (OR) = 3.94, p = 0.003, corrected p (p(c)) = 0.03] and the TNFRSF1A +36AG genotype (OR = 4.81, p = 0.002, p(c) = 0.04) were significantly associated with persistent PR. The frequency of TNFRSF1B +676T/+1663A was increased in PR patients (OR = 2.12, p = 0.01), but failed to reach statistical significance after Bonferroni correction. No correlation was observed between persistent PR and TNFalpha, TNFRSF1A -609G/T, or IL-1beta SNPs. CONCLUSIONS: The results of this study provide evidence of an association between persistent PR and SNPs within the TNFRSF1A gene, and suggest that TNFRI is involved in the aetiopathogenesis of PR.


Assuntos
Polimorfismo de Nucleotídeo Único , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Doenças Reumáticas/genética , Adulto , Idade de Início , Anticorpos Antinucleares/sangue , Mapeamento Cromossômico , Cromossomos Humanos Par 1 , Cromossomos Humanos Par 12 , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Interleucina-1beta/genética , Masculino , Pessoa de Meia-Idade , Peptídeos Cíclicos/sangue , Doenças Reumáticas/sangue , Doenças Reumáticas/classificação , Fator Reumatoide/sangue
5.
Microbes Infect ; 9(1): 1-6, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17196419

RESUMO

Dengue virus (DV) is a flavivirus and its urban transmission is maintained largely by its mosquito vectors and vertebrate host, often human. In this study, investigation was carried out on the involvement of domain III of the envelope (E) glycosylated protein of dengue virus serotypes 1 and 2 (DV-1 and DV-2 DIII) in binding to host cell surfaces, thus mediating virus entry. Domain III protein of flavivirus can also serve as an attractive target in inhibiting virus entry. The respective DV DIII proteins were expressed as soluble recombinant fusion proteins before purification through enzymatic cleavage and affinity purification. The purified recombinant DV-1 and DV-2 DIII proteins both demonstrated the ability to inhibit the entry of DV-1 and DV-2 into HepG2 cells and C6/36 mosquito cells. As such, the DV DIII protein is indeed important for the interaction with cellular receptors in both human and mosquito cells. In addition, this protein induced antibodies that completely neutralized homologous dengue serotypes although not with the same efficiency among the heterologous serotypes. This observation may be of importance when formulating a generic vaccine that is effective against all dengue virus serotypes.


Assuntos
Vírus da Dengue/fisiologia , Proteínas do Envelope Viral/fisiologia , Internalização do Vírus , Animais , Linhagem Celular , Culicidae/citologia , Culicidae/virologia , Vírus da Dengue/metabolismo , Vírus da Dengue/patogenicidade , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Terciária de Proteína , Proteínas do Envelope Viral/metabolismo
6.
Antiviral Res ; 72(3): 216-23, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16870272

RESUMO

RNA interference is one of the effective emerging anti-viral strategies to inhibit virus infection in cells. In this study, a small interfering RNA expressing vector (pSilencer-NS5) targeting the NS5 gene of West Nile virus (WNV) was employed to target and destroy WNV transcripts. Real-time PCR revealed drastic reduction in WNV RNA transcripts in pSilencer-NS5-transfected Vero cells. The virus infectious titre was also significantly reduced by 90% as determined by plaque assays. The resulting decrease in virus replication was shown to be specific since both scrambled and nucleotide(s) mismatch siRNA against WNV NS5 gene did not have any effect on WNV productive yields. Furthermore, Western immunoblot analysis on the expression of viral NS5 and envelope (E) proteins showed significant down-regulation on the expression of viral NS5 and envelope (E) proteins in virus-infected cells that were pre-transfected with pSilencer-NS5. These data clearly supported the notion that the expression of vector-based siRNA against WNV NS5 gene is able to exert its silencing effect on WNV-infected cells without inducing cytotoxicity, hence holding promise in therapeutic treatment of this important emerging infectious disease.


Assuntos
Interferência de RNA , RNA Interferente Pequeno/metabolismo , Proteínas não Estruturais Virais/biossíntese , Replicação Viral , Vírus do Nilo Ocidental/fisiologia , Animais , Chlorocebus aethiops , Citoplasma/química , Expressão Gênica , Vetores Genéticos , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Estabilidade de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Viral/genética , RNA Viral/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Vero , Proteínas do Envelope Viral/biossíntese , Proteínas não Estruturais Virais/genética , Replicação Viral/genética , Vírus do Nilo Ocidental/genética
7.
Int J Clin Pract ; 59(5): 614-6, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15857361

RESUMO

Aneurysm of the superior mesenteric artery (SMA) is rare. We, in this study, present the case of a 21-year-old woman with a history of heroin abuse who was admitted to our hospital for infective endocarditis complicated by floating vegetation at the posterior mitral valve. After receiving 2-week antibiotic treatment, the patient had acute abdominal pain. Computed tomography demonstrated an aneurysm at the SMA. The mycotic aneurysm was resected and the mitral valve was repaired successfully. This report reviews the pathophysiology of mycotic aneurysms of the SMA and role of computed tomography in the differential diagnosis of this condition from acute mesenteric ischaemia.


Assuntos
Aneurisma Infectado/microbiologia , Dependência de Heroína/microbiologia , Artéria Mesentérica Superior/microbiologia , Adulto , Aneurisma Infectado/diagnóstico , Aneurisma Infectado/cirurgia , Ecocardiografia , Endocardite Bacteriana/diagnóstico , Endocardite Bacteriana/microbiologia , Endocardite Bacteriana/cirurgia , Feminino , Humanos , Artéria Mesentérica Superior/cirurgia , Valva Mitral/microbiologia , Insuficiência da Valva Mitral/diagnóstico , Insuficiência da Valva Mitral/microbiologia , Insuficiência da Valva Mitral/cirurgia , Tomografia Computadorizada por Raios X
8.
J Gen Virol ; 86(Pt 2): 405-412, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15659760

RESUMO

The envelope glycoprotein located at the outermost surface of the flavivirus particle mediates entry of virus into host cells. In this study, the involvement of domain III of West Nile virus (WNV-DIII) envelope protein in binding to host cell surface was investigated. WNV-DIII was first expressed as a recombinant protein and purified after a solubilization and refolding procedure. The refolded WNV-DIII protein displays a content of beta-sheets consistent with known homologous structures of other flavivirus envelope DIII, shown by using circular dichroism analysis. Purified recombinant WNV-DIII protein was able to inhibit WNV entry into Vero cells and C6/36 mosquito cells. Recombinant WNV-DIII only partially blocked the entry of dengue-2 (Den 2) virus into Vero cells. However, entry of Den 2 virus into C6/36 was blocked effectively by recombinant WNV-DIII. Murine polyclonal serum produced against recombinant WNV-DIII protein inhibited infection with WNV and to a much lesser extent with Den 2 virus, as demonstrated by plaque neutralization assays. Together these results provided strong evidence that immunoglobulin-like DIII of WNV envelope protein is responsible for binding to receptor on the surface of host cells. The data also suggest that similar attachment molecule(s) or receptor(s) were used by WNV and Den 2 virus for entry into C6/36 mosquito cells.


Assuntos
Antígenos Virais/farmacologia , Proteínas do Envelope Viral/farmacologia , Replicação Viral/efeitos dos fármacos , Vírus do Nilo Ocidental/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Antígenos Virais/biossíntese , Linhagem Celular , Chlorocebus aethiops , Culicidae , Vírus da Dengue/efeitos dos fármacos , Soros Imunes/imunologia , Camundongos , Dados de Sequência Molecular , Testes de Neutralização , Estrutura Terciária de Proteína , Proteínas Recombinantes/farmacologia , Alinhamento de Sequência , Solubilidade , Células Vero , Proteínas do Envelope Viral/biossíntese , Proteínas do Envelope Viral/química , Vírus do Nilo Ocidental/imunologia
9.
J Virol ; 78(19): 10543-55, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15367621

RESUMO

The pathway of West Nile flavivirus early internalization events was mapped in detail in this study. Overexpression of dominant-negative mutants of Eps15 strongly inhibits West Nile virus (WNV) internalization, and pharmacological drugs that blocks clathrin also caused a marked reduction in virus entry but not caveola-dependent endocytosis inhibitory agent, filipin. Using immunocryoelectron microscopy, WNV particles were seen within clathrin-coated pits after 2 min postinfection. Double-labeling immunofluorescence assays and immunoelectron microscopy performed with anti-WNV envelope or capsid proteins and cellular markers (EEA1 and LAMP1) revealed the trafficking pathway of internalized virus particles from early endosomes to lysosomes and finally the uncoating of the virus particles. Disruption of host cell cytoskeleton (actin filaments and microtubules) with cytochalasin D and nocodazole showed significant reduction in virus infectivity. Actin filaments are shown to be essential during the initial penetration of the virus across the plasma membrane, whereas microtubules are involved in the trafficking of internalized virus from early endosomes to lysosomes for uncoating. Cells treated with lysosomotropic agents were largely resistant to infection, indicating that a low-pH-dependent step is required for WNV infection. In situ hybridization of DNA probes specific for viral RNA demonstrated the trafficking of uncoated viral RNA genomes to the endoplasmic reticulum.


Assuntos
Vesículas Revestidas por Clatrina/virologia , Endocitose , Vírus do Nilo Ocidental/fisiologia , Citoesqueleto de Actina/virologia , Animais , Antígenos CD/análise , Transporte Biológico , Proteínas do Capsídeo/análise , Chlorocebus aethiops , Vesículas Revestidas por Clatrina/ultraestrutura , Microscopia Crioeletrônica , Citocalasina D/farmacologia , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/virologia , Retículo Endoplasmático/virologia , Endossomos/virologia , Concentração de Íons de Hidrogênio , Proteína 1 de Membrana Associada ao Lisossomo , Proteínas de Membrana Lisossomal , Lisossomos/virologia , Proteínas de Membrana/análise , Microscopia de Fluorescência , Microscopia Imunoeletrônica , Microtúbulos/virologia , Nocodazol/farmacologia , RNA Viral/análise , Células Vero , Proteínas de Transporte Vesicular , Proteínas do Envelope Viral/análise , Vírion/metabolismo , Replicação Viral , Vírus do Nilo Ocidental/patogenicidade
10.
Int J Clin Pract ; 58(6): 564-7, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15311555

RESUMO

Apoptosis, or programmed cell death, is now recognised as an important cellular event during both normal development and specific disease progression. Apoptosis has been suggested to play a critical role in several cardiovascular diseases, but has not yet been identified as a major influence in primary cardiac tumours. A retrospective review of the achieved material at Chang Gung Memorial Hospital revealed seven patients with cardiac myxoma and one with a tumour originating from the crista terminalis, from January 2002 to December 2002. The medical chart, surgical pathology reports and microscopic slides were available in all cases. All patients, including eight cardiac myxomas and one tumour from crista terminalis, were assessed for apoptosis by terminal deoxynucleotidyl transferase nick-end labelling assay. In this study, apoptosis is well documented in all seven myxoma and has even been reported in tumour from the crista terminalis. Interestingly, apoptosis appears related to the nature of the cell properties rather than the incidence of embolism. In conclusion, apoptosis is important in the progression of the primary cardiac tumours, but the mechanism of cardiac tumour regression still remains uncertain.


Assuntos
Apoptose , Neoplasias Cardíacas/patologia , Mixoma/patologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
11.
Transplant Proc ; 35(8): 3139-41, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14697998

RESUMO

Cyclosporine (CsA), a calcineurin inhibitor, has been associated with endothelial dysfunction in transplant patients. Human and in vitro studies suggest that CsA produces endothelial dysfunction by impairing vascular endothelium-dependent relaxation. However, little is know about the CsA effects to modulate the vasorelaxation after prolonged graft preservation. In this study using a protocol designed to eliminate the influences of infusion pressure and shear stress, we evaluated the effect of CsA on vasorelaxation of coronary and pulmonary arteries after 16-hour University of Wisconsin (UW) solution preservation.


Assuntos
Adenosina , Alopurinol , Vasos Coronários/fisiologia , Ciclosporina/farmacologia , Glutationa , Insulina , Preservação de Órgãos/métodos , Artéria Pulmonar/fisiologia , Rafinose , Vasodilatação/fisiologia , Animais , Vasos Coronários/efeitos dos fármacos , Cães , Modelos Animais , Soluções para Preservação de Órgãos , Artéria Pulmonar/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Vasodilatadores/farmacologia
12.
J Med Virol ; 71(3): 463-72, 2003 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12966555

RESUMO

West Nile (Sarafend) virus has previously been shown to egress by budding at the plasma membrane of infected cells, but relatively little is known about the mechanism involved in this mode of release. During the course of this study, it was discovered that actin filaments take part in the virus maturation process. Using dual-labeled immunofluorescence and immunoelectron microscopy at late infection (10 hr p.i.), co-localization of viral structural (envelope and capsid) proteins with actin filaments was confirmed. The virus structural proteins were also immunoprecipitated with anti-actin antibody, further demonstrating the strong association between the two components. Perturbation of actin filaments by cytochalasin B strongly inhibited the release of West Nile virus (approximately 10,000-fold inhibition) when compared with the untreated cells. Infectious virus particles were recovered after the removal of cytochalasin B. Further confirmation was obtained when nucleocapsid particles were found associated with disrupted actin filaments at the periphery of cytochalasin B-treated cells. Together, these results showed that actin filaments do indeed have a key role in the release of West Nile (Sarafend) virions.


Assuntos
Citoesqueleto de Actina/metabolismo , Membrana Celular/virologia , Vírus do Nilo Ocidental/fisiologia , Animais , Chlorocebus aethiops , Citocalasina B , Humanos , Microscopia de Força Atômica , Microscopia Imunoeletrônica , Células Vero , Proteínas Estruturais Virais/metabolismo
13.
Virology ; 312(2): 458-69, 2003 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-12919750

RESUMO

This study attempts to isolate and characterize West Nile virus-binding molecules on the plasma membrane of Vero and murine neuroblastoma cells that is responsible for virus entry. Pretreatment of Vero cells with proteases, glycosidases (endoglycosidase H, alpha-mannosidase), and sodium periodate strongly inhibited West Nile virus infection, whereas treatments with phospholipases and heparinases had no effect. The virus overlay protein blot detected a 105-kDa molecule on the plasma membrane extract of Vero and murine neuroblastoma cells that bind to WN virus. Treatment of the 105-kDa molecules with beta-mercaptoethanol resulted in the virus binding to a series of lower molecular weight bands ranging from 30 to 40 kDa. The disruption of disulfide-linked subunits did not affect virus binding. N-linked sugars with mannose residues on the 105-kDa membrane proteins were found to be important in virus binding. Specific antibodies against the 105-kDa glycoprotein were highly effective in blocking virus entry. These results strongly supported the possibility that the 105-kDa protease-sensitive glycoprotein with complex N-linked sugars could be the putative receptor for WN virus.


Assuntos
Membrana Celular/metabolismo , Glicoproteínas/metabolismo , Receptores Virais/metabolismo , Vírus do Nilo Ocidental/metabolismo , Animais , Chlorocebus aethiops , Glicoproteínas/antagonistas & inibidores , Glicoproteínas/química , Camundongos , Peso Molecular , Neuroblastoma/metabolismo , Ligação Proteica , Transporte Proteico , Receptores Virais/antagonistas & inibidores , Receptores Virais/química , Células Vero
14.
J Gen Virol ; 83(Pt 10): 2427-2435, 2002 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-12237424

RESUMO

Both polarized epithelial Vero (C1008) and non-polarized Vero (control) cells were grown on permeable cell culture inserts and infected either apically or basolaterally with West Nile (WN) or Kunjin (KUN) virus. KUN virus (closely related to WN virus) was used as a comparison. Using indirect immunofluorescence and plaque assays of productive virus titres, entry of WN and KUN viruses was confined to the apical surface of polarized epithelial cells. For the first time, these results provided evidence on the distribution of flavivirus-specific receptor(s) in polarized epithelial cells; that is to say that receptor expression was shown to be predominant at the apical surface. In addition, the release of these viruses from polarized Vero C1008 epithelial cells was also examined. Egress of WN virus strain Sarafend (S) was observed to occur predominantly at the apical surface of Vero C1008 cells. In contrast, the release of KUN virus was bi-directional from polarized Vero C1008 cells. Furthermore, disruption of the cellular microtubule network was shown to inhibit the apical release of WN (S) virus but had no effect on the release of KUN virus. Hence, the difference in the release of these closely related viruses suggested the involvement of a microtubule-dependent, polarized sorting mechanism for WN virus proteins but not for KUN virus proteins in polarized epithelial cells.


Assuntos
Vírus do Nilo Ocidental/fisiologia , Animais , Membrana Celular/ultraestrutura , Membrana Celular/virologia , Polaridade Celular , Chlorocebus aethiops , Células Epiteliais/virologia , Microtúbulos/virologia , Células Vero , Proteínas do Envelope Viral/metabolismo , Vírus do Nilo Ocidental/metabolismo , Vírus do Nilo Ocidental/ultraestrutura
15.
J Med Virol ; 67(1): 127-36, 2002 May.
Artigo em Inglês | MEDLINE | ID: mdl-11920827

RESUMO

Previous studies have shown that West Nile (Sarafend) virus matured by budding at the plasma membrane, which differs from the usual intracellular maturation of other flaviviruses. The present study investigated the trafficking mechanism of the envelope (E) and capsid (C) proteins of West Nile (Sarafend) virus during the replication cycle. The use of time-based double-immunofluorescence labelling coupled with the Triton X-100 extraction procedure revealed that both the E and C proteins were transported from the perinuclear region towards the plasma membrane along the microtubules simultaneously. The strong association of these virus proteins with the microtubules was demonstrated further with Triton X-100 extraction procedure coupled with double immunogold-labelling. Extraction of infected cells with Triton X-100 in high salt also revealed that virus E proteins were associated with the microtubules via protein-protein interaction. The disruption of microtubules with vinblastine sulphate inhibited the trafficking of both the virus E and C proteins. Both virus structural proteins were observed to co-localise and retained within vinblastine sulphate-induced microtubulin paracrystals. Extracellular virus production was also reduced drastically by vinblastine sulphate at non-cytotoxic concentration. Subsequent studies revealed that the transportation of virus E protein was associated with the microtubules-based motor protein, kinesin.


Assuntos
Capsídeo/metabolismo , Proteínas do Envelope Viral/metabolismo , Vírus do Nilo Ocidental/metabolismo , Animais , Transporte Biológico , Chlorocebus aethiops , Técnica Indireta de Fluorescência para Anticorpo , Cinesinas/metabolismo , Microscopia Eletrônica/métodos , Microscopia de Fluorescência/métodos , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Células Vero , Vimblastina/farmacologia , Proteínas Estruturais Virais
16.
J Med Virol ; 65(4): 758-64, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11745942

RESUMO

It has been difficult to detect and visualize the physical nucleocapsid particles during the replication process of the flaviviruses. The use of cryo-immunoelectron microscopy has clearly revealed the capsid proteins and nucleocapsid particles of West Nile (Sarafend) virus (a flavivirus) for the first time. Physical nucleocapsid particles accumulated in large numbers from 8 hr postinfection. Double immunolabeling of the envelope and capsid proteins showed a close association of these structural proteins for most of the replication cycle. By 10 hr postinfection, budding of nucelocapsids from the plasma membrane was very obvious. Although maturation at the plasma membrane was the dominant mode, during late infection, intracellular maturation into large vacuoles was also observed.


Assuntos
Membrana Celular/virologia , Nucleocapsídeo/fisiologia , Vírus do Nilo Ocidental/fisiologia , Animais , Chlorocebus aethiops , Microscopia Crioeletrônica , Imuno-Histoquímica , Nucleocapsídeo/química , Nucleocapsídeo/ultraestrutura , Proteínas do Nucleocapsídeo/análise , Fatores de Tempo , Células Vero , Proteínas do Envelope Viral/análise , Replicação Viral , Vírus do Nilo Ocidental/ultraestrutura
17.
Chang Gung Med J ; 24(7): 451-4, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11565252

RESUMO

How to optimally treat deep sternal wound infection after open wound infection remains controversial. Biomaterial advances have made local antibiotics-releasing systems a promising alternative for treating deep sternal wound infection. Two patients with deep sternal wound complications were treated with radical wound debridement, sternal refixation, retrosternal suction drainage, bilateral pectoralis major muscle flaps and placement of collagenous drug carriers loaded with vancomycin underneath, above and between the sternal edges. No treatment failure and death occurred in these patients. There were no side effects, treatment failures or deaths after adjuvant treatment with collagenous vancomycin. Preliminary results of these 2 case studies demonstrate the feasibility of successfully treating deep sternal wound infections with collagenous vancomycin in combination with surgical debridement. This technique is easily performed, reliable and safe.


Assuntos
Antibacterianos/administração & dosagem , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Colágeno/administração & dosagem , Esterno/cirurgia , Infecção da Ferida Cirúrgica/tratamento farmacológico , Vancomicina/administração & dosagem , Idoso , Portadores de Fármacos , Humanos , Masculino , Pessoa de Meia-Idade
18.
J Formos Med Assoc ; 100(1): 32-4, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11265257

RESUMO

BACKGROUND AND PURPOSE: Tuberculous (TB) pleurisy results from a delayed hypersensitivity reaction involving macrophages, T-cells, and many cytokines (including tumor necrosis factor, interferon-gamma, and interleukin 1 and 2). Infection by Mycobacterium tuberculosis induces apoptosis in gamma/delta T-cells and macrophages. Fas ligand (FasL) is a type II membrane protein that plays an important role in the regulation of apoptosis and has an intimate relation with these cells and cytokines. A soluble form of FasL (sFasL) exists in a variety of human body fluids, including serum, pleural effusion, cerebral spinal fluid, and ocular fluid. Therefore, we hypothesized that Fas activity is elevated in TB pleurisy. This study investigated the concentration of sFasL in TB pleural effusions and compared it with expression of sFasL in various other pleural effusions. METHODS: Using an enzyme-linked immunosorbent assay, we investigated the sFasL concentrations of 80 pleural effusions from patients with various diagnoses. RESULTS: The median sFasL concentration in the TB pleural effusion group was 104.91 pg/mL (n = 32). This was significantly higher than values in the transudate group (median value, 20.02 pg/mL, n = 9, p < 0.001) and patients with malignant effusion associated with adenocarcinoma of the lung (median value, 23.29 pg/mL, n = 14, p < 0.001). Lymphoproliferative disease could not be distinguished from TB based on sFasL concentrations in pleural effusion. CONCLUSIONS: The sFasL concentration in TB pleural effusions is significantly higher than that in adenocarcinomatous pleural effusions, which are the most common malignant pleural effusions. This difference may serve as a diagnostic tool to differentiate these two most commonly encountered unexplained pleural effusions. Determination of the cellular source and the actual role of the abundant sFasL in TB pleurisy will require further investigation.


Assuntos
Glicoproteínas de Membrana/análise , Derrame Pleural/metabolismo , Tuberculose Pleural/metabolismo , Adenocarcinoma/metabolismo , Idoso , Ensaio de Imunoadsorção Enzimática , Proteína Ligante Fas , Feminino , Humanos , Neoplasias Pulmonares/metabolismo , Masculino , Pessoa de Meia-Idade
19.
Sheng Li Xue Bao ; 53(4): 296-302, 2001 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-11930209

RESUMO

In order to elucidate the intrinsic mechanism underlying proliferation and differentiation of megakaryocytes during ontogenesis, CD34(+) cells were isolated from human fetal liver (FL) with a high-gradient magnetic sorting system (MACS) and were incubated in liquid suspension with 50 and 100 ng/ml of thrombopoietin (TPO) and in MegaCult(Tm) -C semi-solid culture system with 0, 12.5, 25, 50, 100, and 200 ng/ml of TPO. The cell number, colony number of CFU-Mk, platelet-associated antigen phenotype, and DNA ploidy of CD41(+) cells were examined from d 0 to d 12 in culture. The expression patterns of cyclins B1, D1, and D3 were also analyzed by using immunoblot and flow cytometry. TPO stimulated proliferation of CD34(+) cells of FL from 1 x 10(5)/ml to 13.12 +/-4.06 10(5)/ml with 95% of CD41a(+) cells and 3% of CD34(+) cells after 12 d of culture. Most of the megakaryocytes (MKs) derived from FL were in 2 N ploidy class, and few in 4 N ploidy class, but no megakaryocytes ploidy class was higher than 4 N. The effect of TPO on the formation of CFU-Mk colonies from FL derived CD34(+) cells is shown in a dose-response curve. The expression of cyclin B1 increased progressively and the high level of cyclin B1 was maintained in FL CD34(+) cells induced by TPO during 12 d of culture. A high level of cyclin B1 appeared on FL derived MKs of G1 phase at d 12. The expression of cy-of cyclins D1 and D3 gradually increased in FL CD34(+) cells, which was induced by TPO during the initial 6-day incubation. Afterwards, the level of cyclins D1 and D3 decreased progressively, particularly in MKs which were in G2+M phases. These data suggest that (1) TPO induced proliferation and differentiation of FL derived CD34(+) cells through upregulation of cyclin B1 in G2+M phases and cyclins D1 and D3 in all phases of cell cycle, and (2) Continuing high level of cyclin B1 and decreases of cyclins D1 and cyclin D3 on MKs in G2+M phases may contribute to a retardation of MK endoreduplication.


Assuntos
Antígenos CD34/metabolismo , Ciclinas/metabolismo , Fígado/citologia , Megacariócitos/citologia , Trombopoetina/farmacologia , Diferenciação Celular , Divisão Celular , Células Cultivadas , Feto , Células-Tronco Hematopoéticas/citologia , Humanos
20.
Chang Gung Med J ; 23(7): 405-12, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10974755

RESUMO

BACKGROUND: Cost containment is the driving force behind current health care reform. While video-assisted thoracic surgery (VATS) permits a less invasive approach to surgical diseases of the chest, cost is one aspect that is seldom discussed. In Asia, cost-effectiveness essential for the survival of this approach. We present our cost-effective experience with VATS in 2300 patients over a 7-year period. METHOD: Between 1992 and 1999, 2300 patients underwent video-assisted thoracic surgery at Chang Gung Memorial Hospital. The mean age was 53.2 years (range, 22 days to 102 years); 67% (1541 patients) were men. The VATS technique was mainly performed based on traditional surgical principles. Conventional instruments and the fundamental surgical techniques of dissection, suturing, hemostasis, and tissue approximation that are familiar in the open setting were modified and used to enhance cost savings during VATS. RESULTS: Surgery was performed on 41 patients on an emergency basis (24 with impending cardiac tamponade and 17 with hemothorax). The mean hospital stay of the patients treated by VATS was 4.5 days. The majority of the patients were operated on successfully using conventional instruments under video vision. The overall operative cost was decreased as compared to common VATS techniques. No delayed morbidity was noted in our patients after a mean follow-up period of 39 months (range, 1 to 68 months). CONCLUSION: It is our experience that VATS procedures should be performed with the same expertise as open surgery. Conventional instruments similar to those used in open thoracotomy can be incorporated in VATS, and with a more natural hand manipulation. Only through stringent use of expensive endoscopic consumables and application of modified techniques based on traditional surgical principles can VATS be performed efficaciously and economically for a wide range of thoracic conditions.


Assuntos
Cirurgia Torácica Vídeoassistida/economia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade
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