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With the increasing incidence of thyroid cancer worldwide and the increasing demand for surgery, the risk of parathyroid injury is also increasing, which will lead to postoperative hypoparathyroidism (HP) and hypocalcemia. In order to improve the quality of life of patients after surgery, there is an urgent need to develop a novel platform that can identify the parathyroid gland immediately during surgery. The parathyroid gland promotes the increase of blood calcium concentration by secreting parathyroid hormone (PTH). Therefore, in this study, we used the sensitive surface-enhanced Raman spectroscopy (SERS) technique for rapid detection of PTH, while the near-infrared dye IR808 was used for resonance with the incident excitation wavelength of 785 nm to achieve the surface enhanced resonance Raman spectroscopy (SERRS) effect and provide a higher signal. The signal is enhanced by up to 200-fold compared to nonresonant sulfhydryl molecules. In addition, we used an aptamer-antibody mode with low Gibbs free energy (Δ G ) and low dissociation constant ( K d ), using the sandwich method, to achieve specific recognition of PTH. By optimizing the incubation temperature, aptamer dosage, particle concentration, and other key factors, the optimal detection conditions of the system were obtained. The sensitivity of PTH detection was as high as 9.75 pg/mL at a 5 min incubation time. In this work, we achieved an extension from standard products to rats to clinically practical samples. Finally, the eluate samples obtained from the actual clinical operations were compared with the lateral flow biosensor (LFB), chemiluminescence microparticle immunoassay (CMIA), and SERRS, respectively. Finally, using the CMIA results as the reference standard, the SERRS assay achieved a sensitivity of 94% and a specificity of 100%. This study provides new ideas and methods for the refined development of thyroid cancer surgery, improving postoperative safety and quality of life of patients, and promoting the new mode of intraoperative diagnosis and treatment.

The aim of this work is to investigate the assembly of foxtail millet prolamin (FP) with chitosan hydrochloride (CHC) and carboxymethyl-beta-cyclodextrin (CMCD) in acetic acid aqueous solutions. The proportion of acetic acid has a positive impact on the disintegration of FP. With the use of 91.0 % (v/v) acetic acid, FP forms smaller particles of approximately 45 nm (naked FP particles) and 220 nm (FP - CHC - CMCD hybrid particles). In the case of using 61.5 % (v/v) acetic acid, the microstructures of bare FP particles and 570 nm composite FP nanoparticles (NPs) are looser, about 485 nm. Acetic acid inhibits the noncovalent bonds, including the hydrophobic interactions, hydrogen bonding and electrostatic attractions between FP and polysaccharides. Therefore, 3.8 % (v/v) acetic acid can nucleate FP to form more compact FP hybrid particles for delivering curcumin (Cur) with higher encapsulation efficiency, storage stability and release performance, and improve the antibacterial and anticancer activity of Cur., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Acute myocardial infarction (AMI) is one of the most common causes of sudden death in cardiovascular disease, and myoglobin (Myo) is the first protein to be released in the blood after the attack, which is an important biomarker for clinical detection of AMI. The "Golden Rescue Time" for acute myocardial infarction is to intervene within the first 30 min after the attack; therefore, a rapid and accurate Myo detection method is needed urgently. In this study, we designed a combined enzyme-linked immunosorbent assay (ELISA) technique with surface-enhanced Raman scattering (SERS) immunoassay (ELI-SERS), which integrates the small sample volume, ease of operation, and excellent linearity of ELISA while utilizing the SERS technique and selecting the molecule with the Raman signal (IR-808), which is in resonance with the excitation wavelength, for further signal enhancement. The sensitivity of the system was further improved by optimizing the key factors in the assay such as incubation time, particle concentration, and temperature. Compared with the sandwich-structured magnetic bead method, no collection and concentration steps are required, simplifying the operation and ultimately realizing a sensitivity of a 5.3 pg/mL antigen detectable in 6 min. In actual serum samples, we achieve 100% accuracy and sensitivity by adding blockers to exclude the effect of heterophilic antibodies in serum and to reduce false positives in blank samples. We also validated the hepatitis B surface antigen test, demonstrating the universality of our system. Overall, this study designed an ultrasensitive and convenient SERS sensor for the detection of Myo, which extends the practical application of SERS and also contributes methods for the detection of other biomarkers.

Hepatitis B virus (HBV) is a major cause of liver cirrhosis and hepatocellular carcinoma, with HBV surface antigen (HBsAg) being a crucial marker in the clinical detection of HBV. Due to the significant harm and ease of transmission associated with HBV, HBsAg testing has become an essential part of preoperative assessments, particularly for emergency surgeries where healthcare professionals face exposure risks. Therefore, a timely and accurate detection method for HBsAg is urgently needed. In this study, a surface-enhanced Raman scattering (SERS) sensor with a sandwich structure was developed for HBsAg detection. Leveraging the ultrasensitive and rapid detection capabilities of SERS, this sensor enables quick detection results, significantly reducing waiting times. By systematically optimizing critical factors in the detection process, such as the composition and concentration of the incubation solution as well as the modification conditions and amount of probe particles, the sensitivity of the SERS immune assay system was improved. Ultimately, the sensor achieved a sensitivity of 0.00576 IU/mL within 12 min, surpassing the clinical requirement of 0.05 IU/mL by an order of magnitude. In clinical serum assay validation, the issue of false positives was effectively addressed by adding a blocker. The final sensor demonstrated 100% specificity and sensitivity at the threshold of 0.05 IU/mL. Therefore, this study not only designed an ultrasensitive SERS sensor for detecting HBsAg in actual clinical serum samples but also provided theoretical support for similar systems, filling the knowledge gap in existing literature.

Background: Curcumin (CUR) and anthocyanins (ACN) are recommended due to their bioactivities. However, their nutritional values and health benefits are limited by their low oral bioavailability. The incorporation of bioactive substances into polysaccharide-protein composite nanoparticles is an effective way to enhance their bioavailability. Accordingly, this study explored the fabrication of bovine serum albumin (BSA)-fucoidan (FUC) hybrid nanoparticles using a two-step pH-driven method for the delivery of CUR and ACN., Results: Under a 1:1 weight ratio of BSA to FUC, the point of zero charge moved from pH ⁓ 4.7 for BSA to around 2.5 for FUC-coated BSA, and the formation of BSA-FUC nanocomplex was pH-dependent by showing the maximum CUR emission wavelength shifting from 546 nm (CUR-loaded BSA-FUC at pH 4.7) and 544 nm (CUR/ACN-loaded BSA-FUC nanoparticles at pH 4.7) to 540 nm (CUR-loaded BSA-FUC at pH 6.0) and 539 nm (CUR/ACN-loaded BSA-FUC nanoparticles at pH 6.0). Elevated concentrations of NaCl from 0 to 2.5 mol L -1 caused particle size increase from about 250 to about 800 nm, but showing no effect on the encapsulation efficiency of CUR. The CUR and ACN entrapped, respectively, in the inner and outer regions of the BSA-FUC nanocomplex were released at different rates. After incubation for 10 h, more than 80% of ACN was released, while less than 25% of CUR diffused into the receiving medium, which fitted well to Logistic and Weibull models., Conclusion: In summary, the BSA-FUC nanocomposites produced by a two-step pH-driven method could be used for the co-delivery of hydrophilic and hydrophobic nutraceuticals. © 2023 Society of Chemical Industry., (© 2023 Society of Chemical Industry.)

Objective: To detect the SMO mutations in odontogenic keratocyst (OKC) and to explore the mechanism behind. Methods: Patients with OKC who received treatment in the Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology,Peking University, from September 2012 to June 2017 were enrolled. OKC samples from 10 patients diagnosed as naevoid basal cell carcinoma syndrome (NBCCS)-related OKC (4 females and 6 males) and 20 patients diagnosed as sporadic OKC (7 females and 13 males) were collected. Genomic DNAs were extracted from fibrous capsules and epithelial lining respectively. SMO mutations were detected and analyzed by Sanger sequencing. Results: Three SMO mutations were found in one NBCCS-associated OKC who carrying c.2081C>G (p.P694R) mutation) and two sporadic OKC who carrying c.907C>T (p.L303F) mutation and c.1247_1248delinsAA (p.G416E), respectively), among which the first two mutations were novel mutations that had not been reported before. Besides, two mutations in sporadic OKC were not paired with PTCH1 mutations. Conclusions: In addition to PTCH1 gene mutations, SMO gene mutations also exist in OKC which might be related to the development of OKC.

Objective: Kaempferol has been reported to play an anti-tumor role in various human cancers, while its role in gallbladder cancer (GBC) is unclear., Materials and Methods: We found that kaempferol significantly inhibited the growth, invasion and migration, meanwhile induced apoptosis through cells arrested at G0/G1 phase of GBC cell lines, including GBC-SD and SGC996 cells in vitro., Results: Kaempferol promoted the release of cytochrome C from the mitochondria to cytoplasm, the activation of c-caspase-3 and c-caspase-9 and increased the expression levels of pro-apoptotic factor Bax, meanwhile decreased the expression levels of anti-apoptotic factor Bcl-2. In addition, the expression levels of CDK4, CDK6 and cyclin D1, which are members of the CDK4/CDK6/cyclin D1 signaling pathway, were also decreased by kaempferol. Moreover, kaempferol could efficiently prevent tumor progression of GBC in the xenograft in vivo., Conclusions: Our results demonstrated that kaempferol suppressed GBC progression through activation of the CDK4/CDK6/cyclin D1 signaling pathway, suggesting that it might be a potential anti-tumor agent for clinical treatment of GBC.

The kelp grouper Epinephelus moara has high economic value and is popular in fisheries and aquaculture in China. In the previous study, we treated the embryos at 16-22 somite stage at 4 °C, -25.7 °C, -140 °C and -196 °C, and successfully obtained surviving embryos in each group. To better understand the molecular changes affected by the low temperatures, we conducted a comparative transcriptome analysis among embryos exposed at 4 °C for 30 min, embryos exposed at -25.7 °C for 30 min and the control group. qPCR assays were conducted for the validation. Signal transduction pathways were highly enriched for the differentially expressed genes. c-Fos, c-Jun, JunD, GADD45, involved in MAPK signaling pathway, were upregulated when embryos were treated at low temperatures. As immediate early genes, Egr-1a and b, and IER2, that respond quickly to the environment stress, their expression increased as well. Hsp70 showed similar expression pattern as immediate early genes. Meanwhile, transcription factors Sox, HES, TFIID, muscle movement and protein synthesis-related genes were downregulated. Taken together, our findings suggest that cooling disrupts gene expression patterns in E. moara embryos. The differentially expressed genes may be involved in cellular resistance against low temperatures, possibly through neural activation, apoptosis, proliferation, differentiation, cellular recovery and heat shock regulation. This study also provides transcriptome dataset of E. moara embryos exposed to cold temperatures for future studies focusing on the molecular effects of cryopreservation., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Recently, interleukin (IL)-32 has been demonstrated to represent a novel biomarker for evaluating the prognosis of patients with gastric and lung cancer; however, its clinical significance in colon cancer remains unknown. In the present study, the IL-32 expression in 60 patients with colon cancer was examined with an immunohistochemistry assay. IL-32 expression was determined in 37 (61.67%) patients with colon cancer. Additionally, IL-32 was associated with tumor size and Dukes' stage. By using the Kaplan-Meier method, patients with positive IL-32 expression had shorter overall survival time, compared with those with negative IL-32 expression. Multivariate analysis indicated that IL-32 could be an independent prognostic factor in patients with colon cancer; therefore, IL-32 may be a novel prognostic biomarker and therapeutic target for colon cancer.

Embryo cryopreservation is important for long-term preservation of germplasm and assisted reproduction. However, it is still very difficult to obtain viable embryos from cryopreserved fish embryos. In this study, embryos of Epinephelus moara were used to investigate the effects of various cryopreservation methods. Embryos in stages 10 pairs somite (10S), 18 pairs somite (18S), 22 pairs somite (22S), tail-bud (TB), embryo twitching (ET) and pre-hatch (PH) were treated with five-step equilibrium penetration in 40% PMG3T vitrification solution, which contained 15.75% 1,2-propylene glycol, 10.50% Methanol, 8.75% Glycerol and 5.00% Trehalose. We found that 18S, 22S, TB and ET stage embryos had higher survival rates and were more tolerant to the vitrification solution. Five-step equilibrium treatments on the embryos at the tail-bud stage were performed using two vitrification solutions: 40% PMG3T and 40% PMG3S, which consisted of 15.75% 1,2-propylene glycol, 10.50% Methanol, 8.75% Glycerol and 5.00% Sucrose. The embryonic survival rate under PMG3S treatment (63.36%) was significantly higher than PMG3T treatment (43.93%) (P < 0.05). PMG3S and PMG3T with concentrations of 35%, 40% and 45% were tested on tail-bud stage embryos. Higher concentration of the vitrification solution led to significantly lower embryonic survival rate (P < 0.05). The survival rate was 36.79-72.05% in PMG3S, and 37.11-55.18% in PMG3T, and there were non-significant differences in embryonic development and malformation rates among the groups treated with different concentrations. The embryonic normal development rates in PMG3S and PMG3T were 21.27% and 11.04%, and the malformation rates were 36.13% and 31.04%, respectively. The optimum treatment condition was 40 min using 40% PMG3S on embryos at the tail-bud stage. Both PMG3S and PMG3T were used for cryopreserving embryos at 16 pairs somite, tail-bud and ET stage in liquid nitrogen, where we obtained 190 surviving embryos, and 44 fishes underwent normal development and hatched. The survival rate of cryopreserved embryos was 5.15%, the normal development rate was 1.31%, and the malformation rate was 3.66%. We found that PMG3S and PMG3T were effective for cryopreservation of Epinephelus moara embryos. The results provide a foundation for further explorations of fish embryo cryopreservation techniques., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Our previous study found that B cell translocation gene 2 (BTG2) was hyper-methylated and down-regulated in side population (SP) cells of hepatocellular carcinoma (HCC) cell line. However, its clinical significances and biological impacts on HCC SP cells remained unclear., Aims: To investigate the prognostic value of BTG2 gene in HCC and its influences on cancer stem cells (CSCs)-like traits of HCC cell line SP cells., Methods: BTG2 expression in human HCC and adjacent non-cancerous tissues was detected by immunohistochemical staining and quantitative real-time PCR, and also obtained from GEO and TCGA data. Its prognostic values were assessed. Its biological influences on HCC cell line SP cells were evaluated using cell viability, cell cycle, plate clone-forming assay, and chemoresistance in vitro and tumorigenicity in vivo., Results: BTG2 expression was significantly suppressed in human HCC compared to adjacent non-cancerous tissues. BTG2 expression was correlated with TNM stage, tumor size and vascular invasion. Lower expression of BTG2 was associated with poorer overall survival and disease-free survival. In vitro, overexpression of BTG2 substantially suppressed cell proliferation and accumulation of HCC cell line SP cells in G0/G1 phase. Colony formation ability was markedly suppressed by BTG2 overexpression. Moreover, sensitivity of HCC cell line SP cells to 5-fluorouracil was substantially increased by overexpression of BTG2. Furthermore, tumorigenicity of HCC cell line SP cells transfected with BTG2 plasmids was significantly reduced in vivo., Conclusions: BTG2 gene could regulate the CSC-like traits of HCC cell line SP cells, and it represented as a molecular prognostic marker for HCC.

Objective: The aim of this study is to explore the expression and significance of serum HGF and MPO in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) in adults. Method: Ninety-six cases of OSAHS patients diagnosed by PSG were divided into 3 groups: Heavy, medium and light according to the range of AHI; 32 cases of outpatients were healthy persons as the control group. The serum HGF and MPO were determined by ELISA; HGF and MPO were measured after comprehensive treat with CPAP in patients with severe OSAHS, and the correlation between HGF, MPO and PSG were analyzed. Result: ①With the severity of OSAHS patients increased, the serum levels of HGF and MPO increased gradually ( F =119.006, 25.138; P <0.05); The concentration of HGF and MPO in each group showed that there was no significant difference between the two groups compared with the control group ( P >0.05); The rest of the two groups were significantly different ( P <0.05). ②There was no correlation between serum levels of HGF, MPO, BMI and age in OSAHS patients ( P >0.05). The change of serum concentration was positively related to the two. All were negatively correlated with AHI, and positively correlated with LSaO₂ ( P <0.05). ③LSaO₂ of 32 patients with severe OSAHS of has significantly increased after 3 months of comprehensive treatment, AHI and peripheral blood HGF, MPO levels were significantly reduced ( P <0.05). The amylin and myeloperoxidase concentrations of blood after 6 months of treatment were significantly lower than those of 3 months after treatment ( P <0.05), which still increased compared with the control group. Conclusion: Surgery combined with CPAP treatment can significantly reduce the degree of hypoxia and cardiovascular damage in OSAHS patients, which was significantly correlated with the treatment time. Combined detection of serum HGF and MPO concentrations in patients with OSAHS has a certain clinical value in judging the condition and curative effect and evaluating the cardiovascular damage., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To investigate the differences of volume of pharyngeal cavity between normal healthy subjects and patients with severe obstructive sleep apnea hypopnea syndrome(OSAHS)before and after surgery and its application value in assessment of surgical curative effect. Method: Forty-four healthy male adults were included in normal group.Thirty-eight patients with severe OSAHS diagnosed by polysomnography were included in experimental group(OSAHS group). Volume of pharyngeal cavity, oropharynx and laryngopharynx measured by acoustic pharyngealmetry and AHI, obstructive apnea frequency and the lowest oxygen saturation recorded by PSG monitor were all obtained in normal groups and experimental groups before and 3 months after the surgery. All the data are statistically analyzed. Result: Volume of pharyngeal cavity,AHI,obstructive apnea frequency and lowest oxygen saturation of patients in experimental group were all improved after surgery compared with pre-operation( P <0.01). Conclusion: The measurement of pharyngeal cavity volume is helpful to the quantitative analysis of upper airway structure and determine the stenosis location, it is also useful to guide surgery and evaluate the curative effect of the surgery., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Aim: To determine the optimal type of surgery for late-stage gastric cancer with hepatic metastases., Methods: We retrospectively analyzed 49 gastrectomies for late-stage gastric cancer conducted in the First Hospital Affiliated to Henan University of Science and Technology between September 2003 and September 2010. All gastrectomy operations were divided into two groups: radical resection (gastrectomy and simultaneous resection of hepatic metastases, n = 31), and palliative resection (gastrectomy without hepatic resection, n = 18). All 49 patients had chemotherapy catheter implantation in the hepatic artery via the gastroduodenal artery. Postoperative complications and cumulative survival rates of the two groups were compared and analyzed., Results: There was no significant difference in the number of perioperative complications between the radical and palliative resection groups (6 and 3 cases, respectively, P > 0.05). The incidence of long-term complications including ileus (3 in the radical resection and 2 in the palliative resection groups) and anastomosis (2 cases in each group) was not significantly different (P > 0.05). The cumulative survival rate was significantly lower in the palliative resection group (P < 0.05)., Conclusion: Radical gastrectomy with resection of hepatic metastases and hepatoarterial catheter implantation is the recommended surgery for late-stage gastric cancer patients with hepatic metastases.

Objective: To examine the expression of cysteinyl leukotriene receptor-1 (CysLTR-1) and cysteinyl leukotriene receptor-2 (CysLTR-2) in the adenoid tissues from children with adenoid hypertrophy (AH) and to explore the role of leukotrienes in the pathogenesis of AH., Methods: Sixty children with AH who were treated by adenoidectomy and/or tonsillectomy were classified into two groups: simple AH and AH plus allergic rhinitis (n=30 each). Twenty children who underwent tonsillectomy due to recurrent purulent tonsillitis were selected as the control group. The expression of CysLTR-1 and CysLTR-2 in the excised tonsil and/or adenoid tissues was determined by immunofluorescence histochemical labeling and integrated optical density measurement., Results: The expression of CysLTR-1 and CysLTR-2 in the adenoid and tonsil tissues increased significantly in both the simple AH group and AH plus allergic rhinitis group compared with the control group (P<0.01). The expression of CysLTR-1 and CysLTR-2 in the AH plus allergic rhinitis group increased more significantly compared with the simple AH group (P<0.01)., Conclusions: CysLTR-1 and CysLTR-2 are highly expressed in the adenoid tissues from children with AH, suggesting that leukotrienes are involved in the pathogenesis of AH.

Overexpression of interleukin (IL)-35 has been found in a variety of malignancies, but the expression status in gastric cancer has yet to be elucidated clearly. In the present study, positive expression of EBI3 and p35 was 63.3% and 70.0% of cases, respectively. EBI3 expression was strongly related with larger tumor size and invasion depth (P<0.05). Similarly, expression of p35 was also correlated with larger tumor size (P<0.05). These results indicate that IL-35 might be involved in growth of gastric cancer. Interestingly, EBI3 and p35 expressions were positive correlated with Ki-67 expression. Moreover, EBI3 immunoreactivity was associated with Bcl-2 staining. Our data suggest IL-35 is correlated with genesis of gastric cancer by regulating growth and apoptosis.

Background: DNA hypermethylation plays important roles in carcinogenesis by silencing key genes. This study aims to identify pivotal genes in hepatocellular carcinoma (HCC) by DNA methylation microarray and to assess their prognostic values., Materials and Methods: DNA methylation microarray was performed in 45 pairs of HCC and adjacent nontumorous tissues and six normal liver tissues to identify hypermethylated genes in HCC. Potential prognosis-related genes were selected among hypermethylated genes by analyzing influences of methylation levels on disease-free survival (DFS) and overall survival (OS) in 45 patients. Their prognostic values were validated in 154 patients with HCC (including the initial 45 patients) to determine the independent prognostic gene., Results: Altogether, 54 CpG islands in 44 genes were hypermethylated in HCC compared with liver tissues. Among them, methylation levels of ERG and HOXA11 were inversely associated with DFS (both P < 0.050), and methylation levels of EYA4 were inversely related to DFS and OS (both P < 0.050). EYA4 expression was inversely related to tumor size (P < 0.050). Lower EYA4 expression and larger tumor size were independent predictors of both shorter DFS and OS, and higher Barcelona Clinic Liver Cancer (BCLC) staging was an independent predictor of shorter OS (all P < 0.050)., Conclusions: EYA4 functions as a prognostic molecular marker in HCC. Its aberrant hypermethylation and subsequent down-regulation may promote tumor progression.

Annexin A3 has been identified as a novel biomarker in different types of cancers. However, little is known about its clinical significances and and biological roles in gastric cancer. In this study, we assessed annexin A3 expression in 80 patients with gastric cancer and explore its correlation with prognosis Moreover, correlations with Ki-67, Bcl-2 and Bax were also investigated. Expression of annexin A3 was increased in gastric cancer compared with that in normal gastric tissues. Annexin A3 expression was significantly associated with tumor volume and TNM stage (p<0.05). and inversely correlation with prognosis of patients. More interestingly, expression of annexin A3 was positive correlated with Ki-67 and Bcl-2 expression. Our study showed annexin A3 might be a potential prognostic marker for gastric cancer and involved in tumorigenesis by regulating apoptosis and proliferation.

Objective: To evaluate the application value of preoperative ultrasound-guided drawing for locating thyroid micronodule in surgery., Methods: A total of 88 patients (with 137 thyroid micronodules) who underwent thyroid surgery was included in the prospective study. Preoperative thyroid ultrasound was conducted in all patients. Select criteria: the maximum diameter of nodule ≤ 1 cm. All patients were randomly divided into two groups: 46 patients (68 micronoduls) in experimental group with ultrasound-guided drawing location of thyroid micronodule and 42 patients (69 micronoduls) in control group without ultrasound-guided drawing location of thyroid micronodule., Results: All thyroid micronodules of experimental group were found quickly and accurately in surgery, and 4 micronodules in 4 patients of control group were not found in surgery. US examinations 3 months after surgery showed that all micronodules in experimental group were completely removed and 4 micronodules in control group retained., Conclusion: Ultrasound-guided drawing is a useful technique for locating and searching accurately thyroid micronodule in surgery.

Background: DNA methylation plays an important role in maintaining pluripotency and regulating the differentiation of stem cells, but the DNA methylation profile of stem cells in hepatocellular carcinoma (HCC) remains unclear., Aims: To investigate the genome-wide DNA methylation profile of side population (SP) cells of HCC, a special subpopulation of cells enriched with cancer stem cells, by DNA methylation microarray analysis and to analyze the functions and signal pathways of the aberrantly methylated genes in SP cells., Methods: Side population cells were isolated from HCC cell lines Huh7 and PLC/PRF/5 using flow cytometry, and the tumorigenicity of these SP cells was assessed in NOD/SCID mice. The genome-wide DNA methylation status of SP cells and non-SP (NSP) cells was detected and compared by DNA methylation microarray analysis. Genes with differential methylation between SP and NSP cells were further analyzed for their functions and roles in related signaling pathways., Results: Subcutaneous inoculation of 1 × 10(3) SP cells yielded tumors in 60 % NOD/SCID mice, whereas no tumor was developed after the inoculation of 1 × 10(6) NSP cells. Genome-wide DNA methylation microarray analysis showed that 72 and 181 genes were hypermethylated and hypomethylated, respectively, in both Huh7 and PLC/PRF/5 SP cells as compared with their corresponding NSP cells. Analyses of signaling pathways revealed that hypermethylated and hypomethylated genes were related to four and eight pathways, respectively., Conclusions: Hepatocellular carcinoma SP cells possessed a differential DNA methylation status compared with NSP cells, and the differentially methylated genes in SP cells were involved in 12 signaling pathways. Our results provide valuable clues for further investigations in elucidating the importance of epigenetic regulation in sustaining HCC SP cells and tumorigenesis.

Purpose: Sorafenib and S-1 (one mixed formulation containing 5-FU prodrug and dihydropyrimidine dehydrogenase inhibitor) were two effective agents against hepatocellular carcinoma (HCC), but whether they had synergistic effects remained unclear. The present study aimed at evaluating their synergistic effects against HCC and its mechanisms., Methods: Inhibitory effects of sorafenib, 5-FU and their combination on HCC cells PLC/PRF/5 and SK-HEP-1 were evaluated. Expressions of transcription factor E2F-1 and its downstream thymidylate synthetase (TS) in the treated cells were determined using real-time PCR and Western blot. In vivo anti-tumoral efficacy of S-1 plus sorafenib on HCC was evaluated in NOD/SCID mice. E2F-1 and TS expressions in tumors were determined by immunohistochemical staining., Results: Sorafenib inhibited growth of HCC cells in dose-dependent manner, with IC50 of 5.4 ± 0.3 μmol/L for PLC/PRF/5 and 5.3 ± 0.5 μmol/L for SK-HEP-1. Sorafenib (1 μmol/L) enhanced inhibitory efficacy of 5-FU on HCC cells in vitro, dropping IC50 of 5-FU from 167.7 ± 12.1 to 105.4 ± 8.4 μmol/L for PLC/PRF/5 and 115 ± 10.2 to 82 ± 7.4 μmol/L for SK-HEP-1 (both p < 0.01). Sorafenib downregulated E2F-1 and TS expressions on HCC cells, and its combination with 5-FU yielded a synergistic downregulation of TS expression on HCC cells. In NOD/SCID mice with subcutaneously inoculated HCC, sorafenib combined with S-1 yielded greater inhibition on tumor growth and remarkable TS suppression when compared with sorafenib or S-1 alone (all p < 0.05)., Conclusions: Sorafenib enhanced therapeutic efficacy of 5-FU/S-1 against HCC through downregulation of E2F-1 and TS expressions. Sorafenib combined with S-1 might represent as valuable therapeutic regimen against HCC.

Objective: To investigate the advantages and disadvantages of ultracision-harmonic scalpel assisted tonsillectomy by compared with conventional tonsillectomy., Methods: Eighty-eight patients were randomly divided into ultrasonic scalpel group (group A, 42 cases) and control group (group B, 46 cases). The tonsillectomy in group A was performed with ultracision-harmonic scalpel, and in group B, the tonsillectomy was performed by routine method. The surgical time (complete removal of tonsils), blood loss, and postoperative sore throat situation were recorded., Results: Surgical time in group A [(14.7 ± 4.0) min] was shorter than that in group B [(28.9 ± 7.6) min], t = -10.691, P < 0.05. Blood loss in group A [(3.1 ± 1.1) ml] was less than that in group B [(19.0 ± 5.2) ml], t = -19.544, P < 0.05. Postoperative sore throat was less painful in group A than that in group B in 10 hours after surgery, but much painful than group B 3 days after surgery and most patients lasted longer. There were statistical differences (P < 0.05). The average peel off time for the tunica albuginea was 8 days after the operation by using traditional method, by compared with the ultrasonic scalpel method, average time was 11 days, the difference showed statistical significance (t = 5.115, P < 0.05)., Conclusions: Compared with traditional tonsillectomy, ultracision-harmonic scalpel tonsillectomy had shorter operative time, less blood loss and so on, but the sore throat symptoms persisted longer. In addition, the tunica albuginea peeled off later, so avoidance of secondary bleeding caused by improper diet was mandatory.

Objective: The effect of the limb salvage of the treatment of Osteosarcoma Consolidated with Obvious Pathological Fractures is not very well, the purpose of this paper is to track the efficacy of limb salvage treatment when the patients accepted the artery intervention chemotherapy and enclosed 2 clinical case-reports., Methods: From January 2003 to September 2005, 2 clinical cases which one is a male, 29 years old, was confirmed osteosarcoma on the left distal femur, and the other is a female, 15 years old and has osteosarcoma on the right arm with obvious pathological fracture. After receiving arterial chemotherapy pump embedded, then started chemotherapy after the tumor biopsy, and the next process prosthesis replacement in limb salvage surgery after 5 times chemotherapy, follow on the next 5 times chemotherapy after the surgery., Results: With an average follow-up 70 months, there are no postoperative infection and prosthesis loosening found, also didn't detected tumor recurrence and metastasis, and the limb function recovered well., Conclusions: With the effective, neoadjuvant chemotherapy and comprehensive treatment, salvage treatment is not the contraindication of the patients with pathological fractures of combined primary osteosarcoma, and the treatment with long-term follow-up effectively.

Objective: To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd.3 antigen against U14 cervical cancer cell of mice., Methods: Mouse brain microvascular endothelial cell bEnd.3 was cultured and identified for preparation endothelial cell bEnd.3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R₂) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd.3 antigen 4 times in 4 weeks (bEnd.3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD₃+CD₈+ surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot., Results: The expression of VEGF-R₂ and integrin αV gene in bEnd.3 cells were expressed highly. After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd.3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11 ± 0.13) cm³ versus (3.38 ± 0.34) cm³]. The CTL response of spleen lymphocytes in vitro showed that bEnd.3-DC cells could kill bEnd.3 cells, the special lysis rate was more than 60%. The percentage of CD₃+CD₈+ spleen lymphocytes in bEnd.3-DC group [(38.6 ± 0.7)%] was higher than those in other groups (P < 0.05). The titer of serum antibody of bEnd.3-DC group was 1:3200, while it was 1:800 in DC group and there were not any in PBS group. Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd.3 cell in bEnd.3-DC group. Western blot analysis revealed that there were specific bands at 220,000 (VEGF-R₂)., Conclusions: bEnd.3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humoral immunity are induced by bEnd.3-DC antigen which maybe have some antigens in bEnd.3 cells that reacts with endothelial cell proliferation-related antigens.

Aim: To explore the specific immunity of dendritic cell (DC) vaccine loading human umbilical vein endothelial cell (HUVEC) antigen.against U14 cervical cancer of mice., Methods: Primary HUVECs were cultured, identified and made into antigen. The BALB/c mice were immunized with DCs loading HUVEC antigen 4 times a week. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, CTL response of spleen lymphocytes in vitro, the percentage of CD3(+)CD8(+) surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and Western blot., Results: HUVECs with high purity were successfully cultured and by identified by immunocytochemistry and RT-PCR. After the vaccine was injected into mice, the tumors of mice in PBS group grew faster than the those in other groups. The tumors of mice in HUVEC-DC groups grew slowly and disappeared after 2 weeks and The tumors of mice in DC group disappeared after 3 weeks. The CTL response of spleen lymphocytes in vitro showed that HUVEC-DC-T cells could kill HUVEC cells. The percentage of CD3(+)CD8(+) surface markers of spleen lymphocytes in HUVEC-DC group was higher than that in other groups. The titer of serum antibody was 1:800. immunocytochemistry analysis indicated HUVEC-DC group had specific antigen-antibody reaction to HUVECs through and Western blot analysis revealed there were specific bands at 130 KDa and 220 KDa., Conclusion: HUVEC-DC vaccine and DC vaccine can inhibit the tumor growth of U14 cervical cancer of mice. The special cellular and humoral immunity are induced by HUVEC-DC vaccine. Furthermore, some antigens in HUVECs maybe have special immune reaction with integrin alphav and VEGF-R2.

Half-smooth tongue sole (Cynoglossus semilaevis) is an important cultured marine fish as well as a promising model fish for the study of sex determination mechanisms. In the present study, a protocol for artificial gynogenesis of half-smooth tongue sole was developed in order to identify the sex determination mechanism and to generate all-female stock. The optimal UV-irradiation dose for genetically inactivating sea perch spermatozoa was determined to be > or =30 mJ/cm(2). The optimal initiation time for cold shock of gynogenetic embryos was determined to be 5 min after fertilization, while the optimal temperature and treatment duration were determined to be 20-25 min at 5 degrees C. Chromosomes from common diploids, gynogenetic haploids, and diploids were analyzed. WW chromosomes were discovered in some of the gynogenetic diploids. The microsatellite marker was applied to analyze gynogenetic diploid fry. Among the 30 gynogenetic diploid fry, 11 fry contained only one allele, while 19 contained two alleles, which had the same genotype as their mother. The female-specific DNA marker was observed in four individuals out of ten gynogenetic diploid fry. Ploidy analysis of 20 putative gynogenetic fry showed them all to be diploid. Thus, a protocol for the induction of artificial gynogenesis has been developed for the first time in half smooth tongue sole, and the sex determination mechanism in the tongue sole was determined to be female heterogametic with the ZW chromosome.

The prediction of beef tenderness was studied using near-infrared spectroscopy. The absorption spectra of beef samples were collected between 4,000 and 10,000 cm(-1), the maximum shear force of these samples was obtained using the Warner-Bratzler attachment, and subjective judgment for the tenderness grade of beef was studied. Beef samples with the maximum shear force less than 6 kg were regarded as tender, and their tenderness grade was defined as the value of 1. Those with the maximum shear force greater than 9 kg were regarded as tough, and their tenderness grade was defined as the value of 3. And those with the maximum shear force between 6 and 9 kg were regarded as medium, and their tenderness grade was defined as the value of 2. The study shows that the absorption value of tougher beef is generally higher than that of tender beef. Multiple linear regression was used to build the model between the absorption value and tenderness grade. The results give the correlation coefficient r is 0.806. The accuracy of the model for predicting tenderness grade of beef was 84.21% for a validation set including 19 samples. This result indicates that NIR spectroscopy is capable of predicting tenderness grade of beef.