Development and evaluation of a bivalent vaccine based on recombinant newcastle disease virus expressing infectious bursal disease virus VP2L-CH3-CH4 in SPF chickens.

Newcastle disease (ND) and infectious bursal disease (IBD) are two viral infectious diseases that are extremely damaging to the poultry industry and are widespread throughout the world. It is necessary to develop a safe and effective vaccine against IBD and ND because vaccination is an effective preventive measure. It has been discovered that recombinant proteins expressed by an expression system in which a fragment of mammalian Immunoglobulin G (IgG) Fragment crystallizable (Fc) is linked to a segment of a gene have antibody-like properties that increase the exogenous protein's serum half-life. Heavy chain constant region 3 and heavy chain constant region 4 (CH3-CH4) of Avian Immunoglobulin Y (IgY) is structurally very similar to mammalian Ig G Fc. In this study, a bivalent vaccine rClone30-VP2L-CH3-CH4-GMCSF was developed by using NDV rClone30-chGM-CSF vector to produce VP2L-CH3-CH4 fusion protein. The vaccine has been given to 14-day-old specific pathogen free (SPF) free chickens to test whether it has the potential to prevent IBD and ND. Anti-IBDV and anti-NDV antibody levels in serum were evaluated using ELISA and HI, respectively, and the contents of CD4+ T, CD8+ T, and B cells in leukocytes were determined via flow cytometry. The contents and mRNA transcription levels of four inflammatory factors, IL-1ß, IL-4, IFN-γ and chGM-CSF, were detected by ELISA and real-time PCR respectively. The results showed that after vaccination with the rClone30-VP2L-CH3-CH4-GMCSF vaccine, the levels of anti NDV and anti IBDV antibodies in chickens were significantly higher than those of the rClone30 vaccine and commercial vaccines. Meanwhile, the contents and transcription levels of inflammatory factors in chickens inoculated with rClone30-VP2L-CH3-CH4-GMCSF were significantly increased, and the proliferation response of B cells, CD4+ and CD8+ T cells was also stronger. However, the rClone30-VP2L-CH3-CH4-GMCSF vaccine had no significant advantage over the rClone30-VP2L-GMCSF vaccine in any of the above-mentioned features. In summary, rClone30-VP2L-CH3-CH4-GMCSF can stimulate the body to produce a stronger immune response, showing its potential to be considered as vaccine against IBD and ND, but the addition of CH3-CH4 did not improve the vaccine's immune effect as expected. The research lays the foundation for developing vaccines for other infectious viral diseases and avoids a unrealistic vaccine optimization method.

Inhibition of miR-22 enhanced the efficacy of icotinib plus pemetrexed in a rat model of non-small cell lung cancer.

OBJECTIVES: To investigate the role of miR-22 in the efficacy of combined icotinib (BPI-2009H) and pemetrexed (LY-231514) on tumor growth and apoptosis in rats with non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Rats were injected with HCC827 cells, which were transfected with anti-miR-22, followed by the treatment of BPI-2009H and/or LY-231514. MTT assay was used to detect the inhibition rate of HCC827 cells. qRT-PCR was performed to examine miR-22 expression in HCC827 cells and lung tumor tissues. Moreover, immunohistochemistry and Western blotting were performed to detect the related-molecule expressions, while TUNEL staining was used to observe cell apoptosis of lung tumor tissues. RESULTS: MiR-22 expression was decreased in HCC827 cells after the treatment of BPI-2009H or LY-231514 in a dose-dependent manner. Both BPI-2009H and LY-231514 increased the inhibition rate of HCC827 cells, which was enhanced by anti-miR-22 with decreased IC50 values. Furthermore, the decreased expression of miR-22 was found after the treatment of BPI-2009H or/and LY-231514 in lung tumor tissues. In addition, the expressions of PCNA, Ki67, and Bcl-2 were reduced, but Bax and Caspase-3 were increased in treated rats, typically in those rats treated with the combination of anti-miR-22, BPI-2009H, and LY-231514. CONCLUSION: Inhibition of miR-22 could enhance the efficacy of icotinib combined with pemetrexed in rats with NSCLC, providing a new perspective for NSCLC therapy.

Results of surgical treatment for primary malignant melanoma of the esophagus: A multicenter retrospective study.

OBJECTIVE: Few large-sample research data sets exist on long-term survival and prognostic factors among patients with primary malignant melanoma of the esophagus (PMME), a rare malignancy associated with poor outcomes. We sought to investigate postoperative survival and prognostic factors in patients with PMME. METHODS: We retrospectively analyzed long-term follow-up results for patients with PMME who underwent surgery at 10 Chinese centers between January 1998 and January 2018. We performed uni- and multivariate analyses to investigate clinicopathologic factors associated with survival. RESULTS: Median overall survival for the entire group (N = 70 patients) was 13.5 months. Female sex (hazard ratio [HR], 0.352; 95% confidence interval [CI], 0.138-0.900; P = .029), ≥12 lymph nodes dissected (HR, 0.274; 95% CI, 0.133-0.563; P < .001), absence of lymph node metastasis (HR, 0.195; 95% CI, 0.084-0.451; P < .001), and postoperative adjuvant therapy (HR, 0.474; 95% CI, 0.249-0.901; P = .023) were factors of favorable prognosis. Preoperative pathologic diagnosis of PMME was as low as 47.1%. A high proportion of patients had lymph node metastasis, including those with early-stage tumors. Rates of lymph node metastasis were 54.2% (13/24) for pT1, 44.4% (12/27) for pT2, 57.1% (8/14) for pT3, and 100% (5/5) for pT4. Regional lymph node recurrence (N = 43 [61.4%]) was the predominant postoperative pattern of recurrence or metastasis. CONCLUSIONS: Female sex, pN0, increased number of lymph nodes dissected, and postoperative adjuvant therapy were associated with better outcomes among patients with PMME. Preoperative pathologic diagnosis of PMME was low, patients had lymph node metastasis (even those with early-stage tumors), and regional lymph node recurrence was common.

Identification of nine microRNAs as potential biomarkers for lung adenocarcinoma.

Lung cancer is a leading global cause of cancer-related death, and lung adenocarcinoma (LUAD) accounts for ~ 50% of lung cancer. Here, we screened for novel and specific biomarkers of LUAD by searching for differentially expressed mRNAs (DEmRNAs) and microRNAs (DEmiRNAs) in LUAD patient expression data within The Cancer Genome Atlas (TCGA). The identified optimal diagnostic miRNA biomarkers were used to establish classification models (including support vector machine, decision tree, and random forest) to distinguish between LUAD and adjacent tissues. We then predicted the targets of identified optimal diagnostic miRNA biomarkers, functionally annotated these target genes, and performed receiver operating characteristic curve analysis of the respective DEmiRNA biomarkers, their target DEmRNAs, and combinations of DEmiRNA biomarkers. We validated the expression of selected DEmiRNA biomarkers by quantitative real-time PCR (qRT-PCR). In all, we identified a total of 13 DEmiRNAs, 2301 DEmRNAs and 232 DEmiRNA-target DEmRNA pairs between LUAD and adjacent tissues and selected nine DEmiRNAs (hsa-mir-486-1, hsa-mir-486-2, hsa-mir-153, hsa-mir-210, hsa-mir-9-1, hsa-mir-9-2, hsa-mir-9-3, hsa-mir-577, and hsa-mir-4732) as optimal LUAD-specific biomarkers with great diagnostic value. The predicted targets of these nine DEmiRNAs were significantly enriched in transcriptional misregulation in cancer and central carbon metabolism. Our qRT-PCR results were generally consistent with our integrated analysis. In summary, our study identified nine DEmiRNAs that may serve as potential diagnostic biomarkers of LUAD. Functional annotation of their target DEmRNAs may provide information on their roles in LUAD.

Clinical characteristics and survival of lung cancer patients associated with multiple primary malignancies.

OBJECTIVES: To investigate the characteristics and survival of lung cancer patients with additional malignant primary cancers. METHODS: Records of lung cancer patients newly diagnosed in Shanghai Pulmonary Hospital between January 2000 and January 2010 were retrospectively reviewed. Patients with second primary lung cancer and those with lung cancer only were included for detailed analysis. RESULTS: Of 27642 newly diagnosed lung cancer patients, 283 patients (1.02%) suffered previous additional primary cancers. Compared with single primary lung cancer, patients with secondary lung cancer associated other primary cancers were more often women (female to male ratio 1:1.72 vs 1:2.58, P = 0.018), older (64.2 vs 60.5 years old, P<0.001), more squamous cell type (30.7% vs 20.5%, P = 0.004), less small cell (3.9% vs 15.5%, P<0.001) type, at earlier stages (17.7% vs 11.0% for stage I, P = 0.014), and more frequently with family history of cancers (7.8% vs 3.9%, P = 0.038). The most common previous primary cancers observed were colorectal (22.0%), breast (18.4%), gastric (14.4%) and larynx cancers (11.9%). Approximately 42.9% of patients were diagnosed with lung cancer 2 to 6 years after diagnosis of initial primary cancers. The survival of patients with secondary lung cancer associated other malignancies was not significantly different from those with single lung cancer (P = 0.491), while synchronous multiple primary malignancies showed worse prognosis compared with those with metachronous ones or single lung cancer (p = 0.012). CONCLUSION: The possibility of second primary lung cancer should always be considered during the follow-up of related cancer types, especially those with family history of cancers. Patients with secondary lung cancer associated other primary malignancies have non-inferior survival than those with single lung cancer.

Surgical treatment and prognosis of octogenarians with non-small cell lung cancer.

OBJECTIVE: To explore the clinical characteristics, surgical treatment and prognosis of non-small cell lung cancer (NSCLC) among elderly patients over 80 years. METHODS: The clinical data, surgical methods, perioperative management, postoperative complications and prognosis of 52 NSCLC patients aged over 80 years were retrospectively analyzed. RESULTS: Out of 52 cases, 27 had a long-term smoking history (51.9%) and 44 were with other diseases (84.6%). Lobectomy was done in 32 cases (65.4%), sub-lobectomy in 20 cases (38.5%), including pulmonary wedge resection in 16 cases (30.8%) and lung segment resection in 4 cases (7.7%). The postoperative complication rate was 44.2% (23/52); the complication rate after lobectomy was 62.5% (20/32) and that after sub-lobectomy was 25% (5/20), with significant difference between lobectomy and sub-lobectomy (P<0.05). Postoperative mortality was 3.8% (2/52). Pathological TNM staging: I a 27 cases (51.9%), I b 12 cases (23.1%), II a 8 cases (15.4%), II b 3 cases (5.8%) and III a 2 cases (3.8%). The 1-, 3- and 5-year survival rate after operation was 87.1%, 59.8%, 19.1%. The 1-, 3- and 5-year survival rate was 86.0%, 61.8%, 21.5% in the patients after lobectomy; that was 89.0%, 58.3%, 18.7% in the patients after sub-lobectomy, with no significant difference between two surgical methods (P>0.05). CONCLUSIONS: Octogenarians with NSCLC are often afflicted with comorbidity, so perioperative management is more complex. Strictly adhering to indications, surgery is still an important treatment of NSCLC patients over 80.

[Combination of single-port thoracoscopy and laparoscopy for the treatment of esophageal carcinoma: report of 6 cases].

OBJECTIVE: To study the feasibility and early results of radical resection of esophageal carcinoma using single-port thoracoscopy combined with laparoscopy. METHODS: From March 2010 to December 2010, 6 patients with esophageal carcinoma underwent radical resection by single-port thoracoscopy combined with laparoscopy in the General Hospital of People's Liberation Army. With the patients at a supine position, laparoscopy was performed to complete stomach mobilization and abdominal lymph node dissection. Thoracoscopy was then carried out with the patients lying on the left to mobilize the esophagus and dissect thoracic lymph nodes. Finally, the stomach was pulled into the thoracic cavity via the hiatus of the diaphragm to construct a tube-like stomach, which was then anastomosed to the esophagus using the OrVil system. RESULTS: No patient was converted to open surgery during the operation. The total operative time ranged from 200 to 320 min. The mean laparoscopic time was 75(range, 45-90) min, and the mean thoracoscopic time 160(120-240) min. The mean intraoperative blood loss was 220(160-300) ml. The mean lymph node retrieval was 12(9-18). No anastomotic fistula, chylothorax, lung infection were found postoperatively. CONCLUSION: After esophageal resection using single-port thoracoscopic and laparoscopy, reconstruction using OrVil system is safe and feasible.

Predictors of postoperative myasthenic crisis in patients with myasthenia gravis after thymectomy.

BACKGROUND: Thymectomy is considered the most effective treatment in patients with myasthenia gravis. This study aimed to explore the predictors of postoperative myasthenic crisis in patients with myasthenia gravis after thymectomy. METHODS: Clinical records of 243 patients with myasthenia gravis who underwent thymectomy were reviewed retrospectively. The following factors were analyzed in relation to the occurrence of myasthenic crisis after thymectomy: gender, age, duration of symptoms, Osserman stage, history of myasthenic crisis, concomitant diseases, preoperative pyridostigmine dose, preoperative steroid therapy, operation approach, operation time, presence of thymoma, major postoperative complications. RESULTS: Forty-four patients experienced postoperative myasthenic crisis during the first month after thymectomy. Univariate analysis revealed that Osserman stage (RR = 0.0976, P = 0.000), history of myasthenic crisis (RR = 0.2309, P = 0.012), preoperative pyridostigmine dose (RR = 0.4349, P = 0.016), thymoma (RR = 0.0606, P = 0.000), and major postoperative complications (RR = 0.1094, P = 0.000) were significantly related to postoperative myasthenic crisis. Multivariate Logistic regression analysis showed that Osserman stage (IIb + III + IV) (RR = 0.0953, P = 0.000), thymoma (RR = 0.0294, P = 0.000), and major postoperative complications (RR = 0.0424, P = 0.000) independently predict postoperative myasthenic crisis. CONCLUSION: Osserman stage (IIb + IIIb + IV), thymoma and major postoperative complications are independent predictors of postoperative myasthenic crisis in patients with myasthenia gravis who underwent thymectomy.

Diagnostic values of SCC, CEA, Cyfra21-1 and NSE for lung cancer in patients with suspicious pulmonary masses: a single center analysis.

We recruited 805 patients with suspicious pulmonary masses that were identified finally as lung cancer or benign pulmonary masses. The serum levels of four tumor markers, including squamous cell carcinoma antigen (SCC), carcinoembryonic antigen (CEA), cytokeratin 19 fragment antigen 21-1 (Cyfra21-1) and neuron specific enolase (NSE) were tested for every patient. Though receiver operating characteristic (ROC) curves indicated unsatisfactory diagnostic power of those four tumor markers for lung cancer, 37.3% of early-staged lung cancer could be diagnosed just on the combination assays of the four tumor markers, under adjusted cut-off values through our statistical analysis retrospectively.

[Surveillance of echinococcosis in high risk areas of Yixing City from 2008 to 2009].

An investigation was conducted retrospectively for echinococcosis (hydatid disease) in Yixing City from 2008 to 2009, the serum or fecal samples of suspected patients during the past 5 years and host animals were assessed by ELISA. There were 2 cases reported by network and 4 confirmed patients by investigation, and among the total 6 cases, 5 cases may be infected in the locality. The positive rates were 0.52%, 0.14% and 1.08% in adults aged over 20 years, children aged 7 to 12 years (by serum tests) and dogs (by fecal tests) respectively.

Early responses of VEGF during acute lung injury induced by seawater immersion after open chest trauma.

BACKGROUND: Immersion in seawater after open chest trauma may induce acute lung injury. Higher osmotic pressure is one of the main characteristics of seawater. The effects of vascular endothelial growth factor (VEGF) on endothelial cell permeability and proliferation have been demonstrated in studies. The early responses and effects of the VEGF on acute lung injury induced by seawater immersion after open chest trauma (SWI-ALI) are unknown. OBJECTIVE: To investigate the levels of VEGF and its receptors in SWI-ALI and further explore whether the levels of VEGFs are connected with the pathogenesis of SWI-ALI. METHODS: We put dogs into group 'seawater' and group 'control'. The control group only suffered from open chest trauma, whereas the seawater group was exposed to seawater after trauma. The levels of total protein in plasma and bronchoalveolar lavage fluid were measured to calculate the pulmonary permeability index. 0, 2, 4, 6 and 8 h after open chest trauma, the plasma samples were collected to test the levels of VEGFs with ELISA kit. Western blotting and real-time RT-PCR were used to measure the VEGF levels in lung. RESULTS: Compared with control animals, plasma osmotic pressure, VEGF and sVEGFR-1 significantly increased in plasma, while VEGF and VEGFR-2 significantly increased in seawater-immersion lung tissue. The levels of VEGF in plasma were significantly correlated with plasma osmotic pressure and pulmonary permeability index. CONCLUSION: Early release of VEGFs increases pulmonary vascular permeability and partially leads to the development of SWI-ALI. VEGFs may have a crucial role in the early onset of SWI-ALI.

Rhinoplasty in Chinese: management of lower dorsum and bulbous nasal tip.

BACKGROUND: Bulbous nasal tips and lower dorsa are common facial features in Chinese people, and surgery to reshape these is frequently requested. The use of silicone implants in rhinoplasty has been widely used in China for many years, but is not suitable for patients seeking Caucasian tip shapes. The creation of an excessively high tip supported only by a silicone implant inevitably leads to implant extrusion. Although many rhinoplasty techniques have been used in Caucasian patients, there is currently no suitable method for Chinese patients, whose anatomy differs from that of Caucasians. The present study was aimed to investigate the clinical outcome of a novel method of rhinoplasty in Chinese people. METHODS: Eighty patients underwent rhinoplasty using our method between 2002 and 2006. We classified the patients into three types, according to the distance between tip defining points, and used different techniques accordingly. Furthermore, an innovative cartilage carving method and a tip fibro-fatty tissue flap were designed and combined with traditional techniques, such as insertion of silicone implant, cartilage grafts, suture techniques and cephalic trimming to reshape the nasal contours. The followup period was 10 - 60 months (average, 21 months). RESULTS: Remarkable modifications in nasal contours were achieved. No complications developed in any of the 80 patients. Seventy-eight patients were satisfied with the results. The outcomes remained unchanged over time. CONCLUSION: Our method is effective and suitable for the treatment of Chinese patients with lower dorsa and bulbous nasal tips.

[Treatment of scar contracture abnormality on the neck of the children with huge trapezius muscle flap expanded].

OBJECTIVE: To seek a kind of safe and effective method to treat scar contracture abnormality on the neck of the children. METHODS: We take the trunk line of transverse cervical artery as the vessel pedicle, and shift huge trapezius muscle flap expanded to the neck in order to treat scar contracture abnormality on the neck of the children. RESULTS: Clinically, we applied this method to treat ten children who suffered from contracture abnormality on the neck, and not only provide enough skin soft tissue (the length may be beyond the midline) to treat the abnormality, but also make the flap alive, obtain the satisfying postoperative effect. CONCLUSIONS: Based on the warranty of the flap blood supply, this flap is very suitable to treat scar contracture abnormality on the neck of the children.

[Experimental treatment of acute lung injury caused by inundation of thoracic cavity by seawater following open chest wound].

OBJECTIVE: To study the effects of lung protective ventilation and pentoxifylline (PTX) on acute lung injury (ALI) caused by open chest wound with seawater inundation of the thoracic cavity. METHODS: A model of ALI caused by open chest wound and seawater inundation of thoracic cavity was reproduced in dogs. Twenty-four healthy dogs were randomly divided into four groups: no-treatment group (group A), ordinary treatment group (group B), lung protective ventilation treatment group (group C), and lung protective ventilation and PTX treatment group (group D). The parameters of hemodynamics, arterial blood gas analysis, plasma osmotic pressure and serum electrolytes in dogs were determined at 0 and 6 hours after injury and at 2 and 4 hours after treatment. Blood samples and bronchoalveolar lavage fluid (BALF) were collected to assess the changes in cytokines including tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and IL-8. RESULTS: The arterial oxygen partial pressure (PaO(2)) and oxygenation index (PaO(2)/FiO(2)) in group B were still lower than normal values at 2 and 4 hours after treatment, but those parameters in group C and group D distinctly recovered. The parameters of hemodynamics, plasma osmotic pressure and serum electrolytes were all normalized in group B, C and D at 2 and 4 hours after treatment compared with those in group A. The levels of TNF-alpha in peripheral blood in group C and the TNF-alpha and IL-8 levels in peripheral blood and IL-6, IL-8 levels in BALF in group D were significantly lower than those in group A and group B after treatment. The TNF-alpha in peripheral blood and IL-8 levels in BALF in group D were also significantly lower than those in group C after treatment. CONCLUSION: Lung protective ventilation is an effective method in the treatment of ALI caused by open chest wound with inundation of seawater in thoracic cavity. PTX can inhibit inflammatory reaction in the lung and peripheral blood.

[Temporal three-point relief-tension suspension technique for facial rhytidectomy].

OBJECTIVE: To evaluate a facial rhytidectomy technique to reduce the complications of temporal alopecia and incisional scar hyperplasia. METHODS: The 82 aged patients were divided into 2 groups in random way: group A in 46 with the use of the three-point relief-tension suspension technique and group B in 36 as control. In group A, the two point of every relief-tension suture was located respectively in the subcutis of the hairline and fascia under the incision. The parallel three sutures formed a mechanical plane to make the relief suture more strong and permanent. The temporal alopecia and incisional scar was observed with the follow-ups after the operation. RESULTS: With the 82 cases of facial rhytidoplasty, the complications of the temporal alopecia and incisional scars in group A were much less than in group B. CONCLUSIONS: This technique could decrease the complications such as temporal alopecia and incisional scars, and get a good and long term effect of temporal lifting simultaneously.

[Latissimus dorsi transfer to restore thoracic malformation of Poland's syndrome].

OBJECTIVE: To study the methods and points for attention of latissimus dorsi muscle flap transplantation to correct the thoracic malformation of Poland's syndrome. METHODS: From 1995 to 2003, 10 patients were diagnosed of Poland's syndrome with absence of pectoris major muscle in all patients. The latissimus dorsi muscle flap was exposed and transferred through a vertical lateral thoracic cut and a short cut beneath the axillary fold. Reconstruction of the anterior axillary wall is one of the major goals to be achieved in this operation. RESULTS: All of the latissimus dorsi muscle flaps survived. Satisfactory outcomes were achieved after 1-2 years of follow-up. CONCLUSION: The latissimus dorsi muscle flap has a stable and reliable blood supply. It is an ideal muscle flap to restore the thoracic malformation of Poland's syndrome.

[A preliminary approach of covering corrugator with rectangle periosteous flap in rhytidectomy].

OBJECTIVE: To evaluate the aesthetic outcome after application of rectangle periosteous flap to cover corrugator. METHODS: On the basis of regular rhytidectomy, a inter-eyebrow periosteous flap is applied to cover the fascia flap of corrugator by means of turnover. This manipulation helps to separate the skin from the muscle and prevent the re-adherence between the skin and muscle. RESULTS: The approach was applied on 15 cases. The follow-up ranged from 6 months to 1 year. The results were satisfactory. CONCLUSIONS: The method is effective in elimination of inter-eyebrow crease.

[Clinical characteristics and outcome of myasthenia gravis with and without thymoma after operation].

OBJECTIVE: To explore the clinical characteristics and outcome of myasthenia gravis with and without thymoma after operation. METHODS: Two hundred and forty-three patients with myasthenia gravis surgically treated in our department from 1978 to 2003 were studied retrospectively. The clinical characteristics, complication, remission and survival were compared between myasthenia gravis with and without thymoma. RESULTS: The patients of myasthenia gravis with thymoma were significantly older (t = 6.138, P = 0.000), had shorter duration of symptom (t = 3.783, P = 0.000), and also had higher myasthenia crisis rates after operation (chi(2) = 64.77, P = 0.000) than those of myasthenia gravis without thymoma. No differences of Osserman classification was found between the two groups (chi(2) = 7.678, P = 0.104). The complete remission rates and partial remission rates of myasthenia gravis with thymoma were significantly lower than those of myasthenia gravis without thymoma at 1 and 3 years (P = 0.049, P = 0.000; P = 0.015, P = 0.010), but no differences at 5 year (P = 0.457; P = 0.699). The survival rates of MG with thymoma were lower than that of MG without thymoma (Log-rank = 18.58, P = 0.000). CONCLUSIONS: The clinical characteristics are different between myasthenia gravis with and without thymoma. The remission of symptom of myasthenia gravis with thymoma is worse than that of myasthenia gravis without thymoma in the near future, but is similar in the long future. The death rates of MG with thymoma is significantly higher than that of MG without thymoma.