Effect of feeding frequency on the growth performance, carcass traits, and apparent nutrient digestibility in geese.

This study was conducted to determine the effect of feeding frequency on growth performance, carcass traits, and apparent nutrient digestibility in geese from 28 to 70 D of age. In experiment 1, a total of 240 geese were distributed in a completely randomized design into 4 treatments and 6 replicates of 10 birds each. The treatments were free access to the feeder (ad libitum) and access to the feeder 3, 4, and 5 times daily. Geese fed 3 times daily had a lower (P < 0.05) BW, ADG, and ADFI and a higher (P = 0.064) feed conversion ratio (FCR) from 28 to 41 D of age compared with the other groups. Geese fed 4 times daily had a higher (P < 0.05) ADG and ADFI and a lower (P < 0.05) FCR from 42 to 55 D of age compared with ad libitum fed geese. Geese fed 3 times daily had a higher (P < 0.05) ADG from 56 to 69 D of age than geese fed ad libitum and 4 times daily. No differences (P > 0.05) in BW, ADFI, ADG, and FCR were observed between ad libitum and feeding frequency groups from 28 to 69 D of age. Carcass traits and gastrointestinal development were not affected (P > 0.05) by feeding frequency. In experiment 2, the apparent nutrient digestibility in geese from 71 to 77 D of age fed using different feeding frequencies was determined using the total fecal collection method. Feeding frequency did not affect (P > 0.05) the apparent digestibility of DM, CP, crude ash, calcium, phosphorous, or ether extract in geese. Our study demonstrates for the first time that compensatory growth can be gained by enhancing feed intake when a lower feeding frequency is imposed on geese. Both ad libitum feeding and fixed feeding frequency for 3 to 5 times daily are suitable for geese from 28 to 70 D of age to achieve optimum production.

[The indication and surgery scope for laparoscopic extraperitoneal lymphadenectomy in locally advanced cervical cancer].

Objective: To explore the indication and surgery scope for laparoscopic extraperitoneal lymphadenectomy in locally advanced cervical cancer. Methods: Laparoscopic extraperitoneal lymphadenectomy was initially performed on patients with locally advanced cervical cancer. The results of preoperative computed tomography (CT) images and serum level of squamous cell carcinoma antigen (SCC-Ag) were analyzed, and the diagnostic efficiencies of the minimum axial diameter (MAD) of lymph node on CT≥0.5 cm, ≥1.0 cm, serum level of SCC-Ag alone or combined to predict the extraperitoneal lymph node metastases were compared. The high-risk factors of common iliac lymph node (CILN) and para-aortic lymph node (PALN) metastases were also analyzed. Results: The lymph node metastasis rate of 81 patients who received the laparoscopic extraperitoneal lymphadenectomy was 59.3% (48/81). The CILN and/or PALN metastasis rate was 24.7%(20/81), and among them, the MAD of CILN and/or PALN ≥0.5 cm on CT images were in only 7 patients. The threshold of SCC-Ag for evaluating lymph node metastasis was 4.8 ng/ml. The accuracy, specificity and Youden index of SCC-Ag≥4.8+ MAD≥1.0 cm group for predicting lymph node metastasis were 78.3%, 100% and 0.6, respectively, and were significantly higher than 57.9%, 12.1% and 0.1 of MAD≥0.5 cm group, 71.1%, 75.8% and 0.4 of MAD≥1.0 cm group, 65.0%, 57.7% and 0.3 of SCC-Ag≥4.8 ng/ml group and 68.3%, 65.4% and 0.4 of SCC-Ag≥4.8 ng/ml+ MAD≥0.5 cm group (P<0.05). All of the 21 patients in SCC-Ag≥4.8 ng/ml+ MAD≥1.0 cm group were detected to occur lymph node metastases, and the metastasis rate of CILN and/or PALN was 38.1%. While only 4 cases of 17 patients in SCC-Ag<4.8 ng/ml+ MAD<1 cm group were confirmed to occur CILN metastases. The difference of lymph node metastasis rate between these two groups was statistically significant (P<0.001). The pathological type, the number of PLN with MAD≥1.0 cm, at least one of the PLN MAD≥1.0 cm and/or MAD of CILN and/or PALN was 0.5~1.0 cm were associated with the CILN and/or PALN metastases (all P<0.05). Conclusions: Those patients with MAD≥1.0 cm+ SCC-Ag≥4.8 ng/ml and with high-risk factors of CILN and/or PALN metastases should undergo laparoscopic extraperitoneal lymphadenectomy to provide explicit guidance for the subsequent therapy. However, the incidence of lymph node metastasis of patients with SCC-Ag<4.8 ng/ml combined with MAD<1.0 cm is low, therefore these patients can accept concurrent chemoradiotherapy directly.

[Laparoscopic type C1 hysterectomy based on the anatomic landmark of the uterus deep vein and its branches for cervical cancer].

Objective: To introduce the laparoscopic type C1 hysterectomy based on the anatomic landmark of the uterus deep vein and its branched and to evaluate its feasibility and safety for cervical cancer and its effect to bladder function and to provide some reference to simplify the surgical procedures of laparoscopic type C1 hysterectomy. Methods: The clinicopathologic data of the patients with stage ⅠA2~ⅡB cervical cancer and who underwent the laparoscopic C1 hysterectomy based on anatomic landmark of the uterus deep vein and its branches between March 2010 and December 2015 was retrospectively analysed. Results: A total of 99 patients received laparoscopic type C1 hysterectomy based on the anatomic landmark of the uterus deep vein and its branches, in which 93 patients reserved unilateral or bilateral pelvic autonomic nerve successfully, the other 6 patients were transfered to receive type C2 hysterectomy due to adhesions, bleeding or the low possibility of curative resection. The failure rate of the surgery was 6.1% (6/99). The average age of these 93 patients was 44.4±8.2 years (range 25~61 years) and there was one case of stage ⅠA2, 84 stage ⅠB1, 2 stage ⅠB2, 5 stage ⅡA1 and 1 stage ⅡB. The number of patients with squamous cell carcinoma was 67, adenocarcinoma was 19, adenosquamous carcinoma was 3, small cell neuroendocrine carcinoma was 3 and mixed type was 1. The average operation time was 4.1±0.5 h, the average amount of intraoperative blood loss was 103.8±84.0 ml and the mean number of excisional pelvic lymph nodes was 29.7±8.9. There was no patient with positive parametrial margin, positive vaginal margin or intraoperative ureteral injury. The postoperative catheter extraction time was 20.3±8.4 d. The median follow-up time was 20 months (rang 5~44 months), the long-term bladder dysfunction rate was 8.6% (8/93). The numbers of locally uncontrolled and distantly metastasis case were both one and both patients died. The fatality rate were 2.2% (2/93). The two-year disease-free survival and overall survival rate were 97.6% and 96.2%, respectively. Conclusion: Laparoscopic type C1 hysterectomy based on the anatomic landmark of the uterus deep vein and its branches is a safe and feasible treatment method for cervical cancer and it provides a new approach for simplifying the surgical procedures of laparoscopic type C1 hysterectomy.

Treatment and outcome of Stage I Al squamous carcinoma of the cervix: a clinicopathologic study of 346 cases.

OBJECTIVE: To analyze the clinicopathologic feature of Stage I A I squamous carcinoma of the cervix (SCC) and to explore the outcome of different surgical methods. MATERIALS AND METHODS: Clinicopathological data of 346 cases with Stage I Al SCC diagnosed between November 2nd, 1995 and December 31st, 2011 were reviewed and analyzed. RESULTS: As major diagnostic method, 44.5% (154/346) patients accepted cold knife conization (CKC), while 58.1% (201/346) patient took total hysterectomy (TH) as their final surgical methods. The trend in treatment methods from 1995 to 2011 revealed that increasing cases were treated with CKC, modified radical hysterectomy (MRH) obviously reduced, while the proportion treated by TH remained unchanged. Due to a small number of cases receiving vaginal trachelectomy (VT) and radical trachelectomy (RT), the authors did not find any obvious changes. PROGNOSIS: The overall recurrence rate was 1.2% (4/346). The overall survival rates for CKC, VT, TH, MRH, and RT were 100%, 100%, 98.2%, 100%, and 100%, and the difference was not statistically significant (p = 0.819). The incidence rate of LVSI was 4.9% (17/346), the overall survival rates for patients with LVSI and without LVSI were 99.3% and 93.3%, respectively, and there was statistical difference between them (p = 0.003). Univariate analysis showed that only LVSI was an important predictor for survival (p = 0.030). CONCLUSIONS: the treatments for Stage I Al SCC are becoming more conservative, and individualized therapy and more frequent surveillance should be administrated to those patients with LVSI.

Comparison of the efficacy and complications of different surgical methods for cervical intraepithelial neoplasia.

OBJECTIVE: The aim of this was study to offer some reference for the treatment of cervical intraepithelial neoplasia (CIN) by comparing complication rates and treatment failure rates of different surgical methods of CIN. METHODS: 1,256 cases of CIN diagnosed by punch biopsy and pathological confirmation of postoperative specimens between January 2002 and June 2007 were reviewed and analyzed, in which 74 cases underwent the loop electrosurgical excision procedure (LEEP), 869 patients adopted cold knife conization (CKC), 49 patients received vaginal enlarged amputation of cervix, and 264 patients accepted extrafascial hysterectomy. The chi-square test was used to compare the rate of complication and treatment failure of different surgical methods. RESULTS: The rates of surgical complications for LEEP, CKC, vaginal enlarged amputation of the cervix and extrafascial hysterectomy were, respectively, 8.1% (6/74) 6.2% (54/869) 6.1% (3/49) and 2.3% (8/264), but this difference was not statistically significant. The treatment failure incidences for LEEP, CKC, vaginal enlarged amputation of cervix and external fascia hysterectomy were, respectively, 4.1% (3/74), 0.2% (2/869), 0.0% (0/49) and 0.4% (1/264). When comparing among the groups, the treatment failure incidence was higher in LEEP than that in CKC (p = 0.004) and extrafascial hysterectomy (p = 0.034); there was no statistically significant difference between CKC and extrafascial hysterectomy, and no significant difference was revealed between vaginal enlarged amputation of cervix and any other group. CONCLUSION: LEEP, CKC, vaginal enlarged amputation of cervix and extrafascial hysterectomy are all secure and effective procedures for patients with CIN, and patients can make their own individual choice depending on different conditions.

Glowing zebrafish: integration, transmission, and expression of a single luciferase transgene promoted by noncovalent DNA-nuclear transport peptide complexes.

The development of vehicles driving foreign DNA into the cell nucleus is essential for effective cellular gene transfer applications. We report that noncovalent binding of nuclear localization signal (NLS) peptides to plasmid DNA enhances nuclear uptake of the DNA and promotes germline integration, inheritance, and expression of a single copy of a luciferase reporter gene in zebrafish. As few as 10 DNA-NLS complexes (0.06 fg plasmid DNA) cytoplasmically injected are sufficient to produce germline-transgenic zebrafish bearing a single copy of the transgene. This corresponds to a 10(5)-fold reduction in DNA concentration compared to commonly used procedures. Use of 10(3) or 10(4) DNA-NLS complexes augments the number of transgene integrations, which occur mostly within 1-4 distinct insertion sites in the genome. In situ hybridization analyses and transmission studies show that transgene integration into the germline and somatic tissues is mosaic, and that the extent of mosaicism is negatively correlated with the amount of DNA-NLS injected. In addition, a larger proportion of zebrafish harboring a single copy of the transgene expresses luciferase, albeit at a 10-fold lower level than those containing numerous transgene insertions. The data demonstrate the potential use of nuclear targeting peptides noncovalently bound to vector DNA to enhance the efficiency of biotechnological nonviral gene transfer applications.

Active transgenes in zebrafish are enriched in acetylated histone H4 and dynamically associate with RNA Pol II and splicing complexes.

We have investigated the functional organization of active and silent integrated luciferase transgenes in zebrafish, with the aim of accounting for the variegation of transgene expression in this species. We demonstrate the enrichment of transcriptionally active transgenes in acetylated histone H4 and the dynamic association of the transgenes with splicing factor SC35 and RNA Pol II. Analysis of interphase nuclei and extended chromatin fibers by immunofluorescence and in situ hybridization reveals a co-localization of transgenes with acetylated H4 in luciferase-expressing animals only. Enrichment of expressed transgenes in acetylated H4 is further demonstrated by their co-precipitation from chromatin using anti-acetylated H4 antibodies. Little correlation exists, however, between the level of histone acetylation and the degree of transgene expression. In transgene-expressing zebrafish, most transgenes co-localize with Pol II and SC35, whereas no such association occurs in non-expressing individuals. Inhibition of Pol II abolishes transgene expression and disrupts association of transgenes with SC35, although inactivated transgenes remains enriched in acetylated histones. Exposure of embryos to the histone deacetylation inhibitor TSA induces expression of most silent transgenes. Chromatin containing activated transgenes becomes enriched in acetylated histones and the transgenes recruit SC35 and Pol II. The results demonstrate a correlation between H4 acetylation and transgene activity, and argue that active transgenes dynamically recruit splicing factors and Pol II. The data also suggest that dissociation of splicing factors from transgenes upon Pol II inhibition is not a consequence of changes in H4 acetylation.