[Optimal melanin removal methods for HE staining, immunohistochemistry and molecular detection].

Objective: To seek the optimal melanin-removal method for hematoxylin and eosin (HE) staining, immunohistochemistry and molecular detection. Methods: Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital, Fuzhou, China between January 2018 and March 2022 were collected and used to make a tissue microarray. Melanin in these cases was removed using warm hydrogen peroxide, double oxidation depigmentation, modified potassium permanganate-oxalic acid or trichloroisocyanuric acid, followed by HE staining. The cases were divided into two cohorts: one was subject to the one of the above four methods to remove melanin first, followed by immunohistochemistry (SOX-10, Ki-67, HMB45 and Melan A), while the other was subject to immunohistochemical staining first and then a melanin removal. Following that, seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above. DNA extraction was then done, followed by assessments of DNA content and quality. Moreover, the completeness of melanin removal, the effect on HE and immunohistochemical staining, and the quality of DNA were compared between the depigmented methods. Results: Regarding the effectiveness of melanin removal, the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective, and both showed residual melanin in only 5.26% (2/38) of the cases. The trichloroisocyanuric acid method showed residual melanin in 10.53% (4/38) of the cases. The worst was the double oxidation depigmentation method, which showed pigment residue in 15.79% (6/38) of the cases. For HE staining, the percentage of good staining with the warm hydrogen peroxide method was 92.11%, higher than the other three methods. For immunohistochemical staining, the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid, double oxidation and trichloroisocyanuric acid were 20.84, 26.63 and 35.02, respectively. These immunohistochemical staining scores were higher than those of melanin removal first followed by immunohistochemistry (8.70, 15.41 and 21.22, respectively). The mean staining score of melanin removal by warm hydrogen peroxide method followed by immunohistochemistry was 33.57, superior to that of immunohistochemistry followed by the melanin removal (19.96). Moreover, the staining scores of HMB45, MelanA and Ki-67 with immunohistochemical staining followed by trichloroisocyanuric acid method were 36.45, 33.79, and 36.24, respectively, while the staining score of SOX10 with melanin removal by warm hydrogen peroxide followed by immunohistochemistry was 34.39. The DNA was significantly degraded by modified potassium permanganate-oxalic acid, double oxidation depigmentation and trichloroisocyanuric acid, whereas the mean concentration of DNA extracted after melanin removal by hydrogen peroxide method was 59.59 µg/L, substantially higher than that of DNA extracted without melanin removal (30.3 µg/L, P=0.001). The A260/A280 of DNA extracted after melanin removal by hydrogen peroxide was between 1.8 and 2.0 in all cases, and the A260/A230 was above 2.0 in sixteen cases, suggesting high purity of DNA. However, the DNA extracted without removing the melanin showed poor purity, with A260/A280 below 1.8 in eight cases and A260/A230 below 2.0 in sixteen cases. Conclusions: Warm hydrogen peroxide showed the least melanin residue, superior HE staining and a minimal effect on DNA purity/quality compared to the other three methods. It thus appears most suitable for PCR, NGS and other molecular detection. Melanin removal with trichloroisocyanuric acid after immunohistochemical staining has the least melanin residual, and thus could be the most convenient and efficient. However, it is noted that the efficacy of the same depigmentation method varies with different antibodies. Therefore, the optimal depigmentation method should be selected based on the specific markers of interest.

Retraction Note: Study on the functions and mechanism of immune functions of human telomerase reverse transcriptase regulating dendritic cells treating sepsis.

The article "Study on the functions and mechanism of immune functions of human telomerase reverse transcriptase regulating dendritic cells treating sepsis", by H.-M. Chen, L.-Q. Wang, H.-P. Wan, H.-Z. Wei, L.-C. Ke, C.-Y. Liu, Q.-Y. Tan, published in Eur Rev Med Pharmacol Sci 2016; 20 (21): 4500-4507-PMID: 27874963 has been retracted by the Editor in Chief for the following reasons. Some concerns were raised on PubPeer (https://pubpeer.com/publications/3604386A706802443E51758A893D6F) about Figures 3, 4, and 5 showing some overlaps and similar bands in Western blots figures. Furthermore, there is a lack of information regarding the ethics approval for the study involving rats. The journal contacted the authors to request the original raw data and information regarding the ethical approval of the manuscript but never received a reply. Therefore, due to major concerns detected, the Editor in Chief mistrusts the results presented and decided to withdraw the manuscript. The corresponding author has been informed about the retraction. This article has been retracted. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/11476.

[Diagnostic value of apparent diffusion coefficient histograms using multiplexed sensitivity encoding diffusion-weighted imaging for distinguishing nasopharyngeal carcinoma from benign hyperplasia].

The data of 115 patients with nasopharyngeal masses (78 males and 37 females) aged between 12 and 78 years at the Sun Yat-sen University Cancer Center from May 2022 to July 2023 were retrospectively reviewed, including 70 cases of nasopharyngeal carcinoma and 45 cases of benign hyperplasia. The mean, median, and percentiles (10th, 25th, 75th, and 90th) of the apparent diffusion coefficient (ADC) histogram derived from multiplexed sensitivity encoding diffusion-weighted imaging (MUSE-DWI) of the benign hyperplasia group were significantly higher than those of the nasopharyngeal carcinoma group (all P<0.05). Conversely, the kurtosis and skewness of benign hyperplasia group were significantly lower than those of the nasopharyngeal carcinoma group (both P<0.05). The area under receiver operating characteristic (ROC) curve of the combined ADC histogram parameters was 0.812 (95%CI: 0.732-0.892), and the sensitivity, specificity and accuracy were 92.86%, 57.78% and 79.13%, respectively. The current study indicates ADC histogram parameters derived MUSE-DWI exhibit significant discriminatory value between nasopharyngeal carcinoma and benign hyperplasia.

[Analysis of developmental coordination disorder in Chinese children].

Objective: To investigate the prevalence of children's developmental coordination disorder (DCD) and its distribution based on different family socioeconomic characteristics in China, in order to provide a theoretical basis for early prevention, diagnosis, and intervention for DCD. Methods: From June to October, 2016, 1 887 children aged 3-10 years from 20 kindergartens and 10 elementary schools from 8 cities in seven geographic areas of China using a stratified cluster sampling method were recruited. With a cross-sectional design, parents were asked to report on their basic information. Children' s motor ability was assessed using the movement assessment battery for children-second edition (MABC-2). Children were grouped by age, sex, body mass index (BMI), one-child status, and family structure. Chi-square test and one-way ANOVA were used to compare family socioeconomic characteristics of children between different groups. Results: Among the 1 887 children, there were 1 110 (58.8%) preschool children (3-6 years of age) and 777 (41.2%) school-aged children (7-10 years of age). There were 982 males (52.0%) and 905 females (48.0%). A total of 5.5% (104 cases) children were diagnosed with DCD, 10.4% (197 cases) with suspected DCD, and 84.1% (1 586 cases) as typical motor developing children. There were no significant differences in prevalence of diagnosed and suspected DCD among different regions (χ²=17.342 and 4.877, P=0.173 and 0.560), total motor coordination score (F=2.759, P<0.05), and the scores of all dimensions (manual dexterity, positioning and grabbing, balance: F=9.276, 5.277, 3.706, all P<0.01). The prevalence of DCD in preschool children was significantly higher than that in school-age children (χ²=11.891, P<0.01). Girls were significantly better than boys in total motor coordination, manual dexterity, and balance (all P<0.01). Boys were significantly better than girls in positioning and grabbing (P<0.01). The prevalence of DCD in boys was significantly higher than that in girls (70 boys (7.1%) and 34 girls (3.8%), χ²=28.508, P<0.01). The total motor coordination ability, manual dexterity and balance of children who are overweighted (BMI>18 kg/m2) were significantly lower than those of children of normal weight (BMI≤18 kg/m2) (all P<0.01). The prevalence of suspected DCD children who are overweighted was significantly higher than that of children of normal weight (χ²=4.369, P<0.05). The difference of total motor coordination ability (F=6.811, P<0.01) and the prevalence of DCD (χ²=14.902, P<0.01) among different family structures were statistically significant. The total motor coordination ability and balance ability of children from multi-child family were better than those of children from one-child family (both P<0.05). Conclusion: The motor coordination ability of Chinese children is well-developed, with differences among different regions, gender, age, BMI, and family structure.

Near-GeV Electron Beams at a Few Per-Mille Level from a Laser Wakefield Accelerator via Density-Tailored Plasma.

A simple, efficient scheme was developed to obtain near-gigaelectronvolt electron beams with energy spreads of few per-mille level in a single-stage laser wakefield accelerator. Longitudinal plasma density was tailored to control relativistic laser-beam evolution, resulting in injection, dechirping, and a quasi-phase-stable acceleration. With this scheme, electron beams with peak energies of 780-840 MeV, rms energy spreads of 2.4‰-4.1‰, charges of 8.5-23.6 pC, and rms divergences of 0.1-0.4 mrad were experimentally obtained. Quasi-three-dimensional particle-in-cell simulations agreed well with the experimental results. The dechirping strength was estimated to reach up to 11 TeV/mm/m, which is higher than previously obtained results. Such high-quality electron beams will boost the development of compact intense coherent radiation sources and x-ray free-electron lasers.

Investigation of the feasibility of synthetic MRI in the differential diagnosis of non-keratinising nasopharyngeal carcinoma and benign hyperplasia using different contoured methods for delineation of the region of interest.

AIM: To assess the feasibility and preliminary diagnostic performances of relaxation times derived from synthetic magnetic resonance imaging (syMRI) for differentiating nasopharyngeal carcinoma from nasopharyngeal benign lymphoid hyperplasia, and to assess the influence of tissue segmentation method on relaxation estimates. MATERIALS AND METHODS: Fifty participants with nasopharyngeal carcinoma (NPC) and 40 participants with benign hyperplasia (NPH) who underwent syMRI examination were enrolled prospectively. T1, T2, and proton density (PD) values were obtained from four different regions of interest (ROIs), namely, partial-section, single-section, three-sections, and whole-lesion. The metrics between NPC and NPH or among different ROIs were compared using Student's t-test or one-way ANOVA. The area under curve (AUC) was calculated to assess the performance of metrics obtained from different ROIs to differentiate NPC and NPH. RESULTS: The T1, T2, and PD values for NPH were significantly higher than those for NPC, regardless of the type of ROI used, except for the PD value obtained from the whole-lesion ROI. The T2 values obtained from the single-section ROI showed the highest diagnostic accuracy in distinguishing NPC from NPH, with an AUC of 0.894, sensitivity of 0.900, and specificity of 0.800. Additionally, the T1, T2, and PD values for nasopharyngeal lesions showed no statistical difference among different kinds of ROI, except for the difference in T1 value between partial-section and other methods. CONCLUSION: Quantitative analysis of syMRI has the potential to distinguish NPC from NPH. Moreover, different types of ROI showed limited influence on the relaxation time estimation for nasopharyngeal lesions.

Age distribution of human papillomavirus infection and neutralizing antibodies in healthy Chinese women aged 18-45 years enrolled in a clinical trial.

OBJECTIVES: Data from clinical trials of human papillomavirus (HPV) vaccines showed that women naïve (negative for both type-specific antibodies and DNA) to vaccine types would derive benefit from vaccination; therefore, an understanding of the proportion of naïve women in different age groups is important for developing HPV vaccination strategies. METHODS: From November 2012 to April 2013, a total of 7372 healthy women aged 18-45 years were recruited in five provinces in China. Cervical specimens and serum samples were collected for each woman at entry. Cervical specimens were first tested by the HPV DNA enzyme immunoassay method; if positive, the specimens were then tested by reverse hybridization line probe assay and HPV-16 and HPV-18 specific polymerase chain reactions. Neutralizing antibodies against HPV-16 or HPV-18 were tested with a pseudovirion-based neutralization assay. RESULTS: The overall prevalence of high-risk HPV DNA was 14.8% (1088/7367, 95% CI 14.0-15.6), and the seroprevalence of neutralizing antibodies against HPV-16 and HPV-18 was 12.6% (925/7367) and 4.9% (364/7367), respectively. In younger women (18-26 years) and middle-aged women (27-45 years), 83.8% (3116/3719) and 81.4% (2968/3648) were naïve to both HPV-16 and HPV-18 (both neutralizing antibodies and DNA were negative), respectively. In addition, 98.5% (3664/3719) and 98.0% (3575/3648) of the younger or middle-aged women were naïve to at least one HPV type (HPV-16 or HPV-18). DISCUSSION: This study revealed that the majority of Chinese women aged 18-26 years and 27-45 years were naïve to both HPV-16 and HPV-18 and would thus derive full benefit from bivalent HPV vaccination.

Temperature and humidity associated with increases in tuberculosis notifications: a time-series study in Hong Kong.

Previous studies have revealed associations of meteorological factors with tuberculosis (TB) cases. However, few studies have examined their lag effects on TB cases. This study was aimed to analyse nonlinear lag effects of meteorological factors on the number of TB notifications in Hong Kong. Using a 22-year consecutive surveillance data in Hong Kong, we examined the association of monthly average temperature and relative humidity with temporal dynamics of the monthly number of TB notifications using a distributed lag nonlinear models combined with a Poisson regression. The relative risks (RRs) of TB notifications were >1.15 as monthly average temperatures were between 16.3 and 17.3 °C at lagged 13-15 months, reaching the peak risk of 1.18 (95% confidence interval (CI) 1.02-1.35) when it was 16.8 °C at lagged 14 months. The RRs of TB notifications were >1.05 as relative humidities of 60.0-63.6% at lagged 9-11 months expanded to 68.0-71.0% at lagged 12-17 months, reaching the highest risk of 1.06 (95% CI 1.01-1.11) when it was 69.0% at lagged 13 months. The nonlinear and delayed effects of average temperature and relative humidity on TB epidemic were identified, which may provide a practical reference for improving the TB warning system.

[Expression and clinical significance of SSH1 in gastrointestinal stromal tumors].

Objective: To investigate the correlations between the expression of the slingshot-1(SSH1) and the general clinicopathological factors in gastrointestinal stromal tumor (GIST) patients. Methods: Immunohistochemical staining was used to detect the expression of SSH1 protein in 96 gastrointestinal stromal tumors, and the correlations between expression level of SSH1 and general clinicopathological factors were analyzed. Results: Totally, there were 96 cases of GIST patients (49 males and 47 males), with a mean age of (55.80±12.33) years. The tumors were located in different organs: 62 cases (64.6%) in the stomach, 29 cases (30.2%) in the intestine, and 5 cases (5.2%) in other organs, with a mean tumor size of (5.11±3.73) cm. The low risk group contained 57 cases (59.4%), 18 cases (18.8%) in middle risk group and 21 cases (21.9%) in high risk group. The pathological examination showed that the positive expression rate of CD117, CD34, DOG-1, Ki-67, SMA, S-100 and Desmin was 85.4%, 84.4%, 86.5%, 90.6%, 10.4%, 3.1% and 2.1%, respectively. In 96 cases of GIST, 37 cases (38.5%) had high expression of SSH1, and SSH1 expression level was correlated with the tumor invasion depth, risk classification, tumor mitotic figure count, CD34(+) (P<0.05). Conclusions: SSH1 plays an important role in the development of GIST. High expression of SSH1 is closely related to the invasion depth, risk classification and tumor mitosis of GIST.

[Differential expression and clinical significance of calretinin in total colonic aganglionosis].

Objective: To evaluate the differential calretinin immunostaining in different segments of total colonic aganglionosis and its utility in the diagnosis. Methods: Nine specimens including ileum and colon segments were obtained from 9 patients with total colonic aganglionosis (TCA), from 2010 to 2016 year, in Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology. Another 9 ganglionic specimens including the same segments from patients with non-Hirschsprung disease (non-HD) patients were collected as control. All cases were immunostained with calretinin. The patterns of calretinin immunostaining were observed, and morphometric analysis of each sample was performed by image analysis program (Image-Pro-Plus). The mean absorbance was evaluated by calculating the areas of the lamina propria occupied by the positively stained area of the calretinin at high power field. Results: The same pattern of calretinin immunostaining was seen in ganglionic ileum and ganglionic colon segments, with staining seen in intrinsic nerves fibers (INF), and in granular aggregates in the lamina propria and muscularis mucosae. There was no significant difference in the numbers of calretinin-positive INF from the ganglionic segments. In contrast, the number of calretinin-positive INF and granular aggregates in aganglionic segments were significantly lower than those in the ganglionic group (P<0.01). In the ileum transitional zone, scattered calretinin staining was observed, and the amount of calretinin-positive INF was significantly lower than those in the proximal segment of ganlionic ileum (P<0.01). Conclusions: Since there is significant different expression of calretinin among the different segments from TCA, calretinin immunostaining has potential value in detecting TCA. It could be an important adjunctive method in detecting TCA in the future.

High expression of long non-coding RNA ATB indicates a poor prognosis and regulates cell proliferation and metastasis in non-small cell lung cancer

Background and aim. Long non-coding RNAs (lncRNAs) have been demonstrated to act as a critical regulator in the processes of tumor biology. In this study, whether lncRNA-ATB is a potential indicator for non-small cell lung cancer (NSCLC) was investigated and its biological function in NSCLC was also determined. Methods. The expression levels of lncRNA-ATB in NSCLC tissues and cell lines were measured. A549 cell line was explored to investigate the functions of lncRNA-ATB in NSCLC. Results. Real-time PCR results showed that lncRNA-ATB expression was up-regulated in both in NSCLC tissues and cell lines. High lncRNA-ATB expression in tumor tissue was associated with larger tumor size, lymph node metastasis, and distant metastasis in patients with NSCLC, respectively. In addition, the patients with high expression of lncRNA-ATB presented a lower survival probability. In vitro experiments showed that down-regulation of lncRNA-ATB promoted the cell apoptosis, whereas inhibited the cell viability, cell migration, and cell invasion. Conclusion. High expression of lncRNA-ATB indicated a poor prognosis and led to the cell proliferation and metastasis in NSCLC (AU)

No disponible

[Pharmacokinetics of praziquantel injection in healthy buffaloes].

Objective To investigate the pharmacokinetics and relative bioavailability of praziquantel injection in buffaloes in contrast to praziquantel tablet. Methods A single oral administration of praziquantel tablet at a dose of 20 mg/kg or intramuscular administration of praziquantel injection at a dose of 10 mg/kg was performed in six healthy adult buffalos according to a twoperiod crossover design. The praziquantel concentration in plasma was determined by a high performance liquid chromatography (HPLC) method. The pharmacokinetic parameters were calculated by non-compartmental analysis. Results The main pharmacokinetic parameters of praziquantel tablet were as follows: Tmax = (0.60±0.29)h, Cmax = (0.57±0.37)µg/ml, T1/2ß = (0.70±0.42)h, AUC = (0.80±0.70) (µg/ml)·h. The main pharmacokinetic parameters of praziquantel injection were as follows: Tmax = (0.65± 0.49)h, Cmax = (3.82±1.17)µg/ml, T1/2ß = (1.00±0.73)h, AUC = (1.61±0.89) (µg/ml)·h. The relative bioavailability of praziquantel injection was 402.5% in contrast to praziquantel tablet. Conclusion The praziquantel injection has pharmacokinetic characteristics of rapid absorption, high bioavailability and extensive distribution, and the clinical recommended dosage of praziquantel injection is 10 mg/kg.

A survey on parasites in wild rodents in Xiji County, a northwestern part of China.

Rodents act as an indicator for evaluation of environment contaminations and public health risks caused by parasites. A survey of parasites in wild rodents was conducted in 14 villages in Xijin County, where alveolar echinococcosis by Echinococcus multilocularis is epidemic. In total, 72 rodents including 25 mice, 16 Citellus dauricus (Daurian ground squirrel), 12 squirrels and 19 mole rat (Myospalax fontanieri) were captured. Infections (2.8%) of Taenia taeniaeformis, which is transmitted mainly between mice and cats, were found in mice in Wangping (WP) and Miaoping (MP) villages, but other cestodes' infections were not observed. WP and MP isolates were principally similar in morphology but, unlike WP isolate, MP isolate had no hooks on the scolex. Using 18S rRNA as a biomarker, the phylogenetic analysis showed that WP and MP isolates grouped together with European and Asian isolates and formed a separate cluster. These results highlights the prevalence of T. taeniaeformis in cats or/and dogs and a risk of opportunistic infections in human populations.

Preoperative NLR and PLR in the middle or lower ESCC patients with radical operation.

Neutrophil lymphocyte ratio (NLR) and platelet lymphocyte ratio (PLR) had been analysed in many kind of tumours, but its role of predict the oesophageal squamous cell carcinoma (ESCC) patients' prognosis was not reach a consensus. Relationship between NLR, PLR and ESCC located in the middle or lower segment was evaluated. 317 patients with ESCC who underwent attempted curative oesophagectomy were analysed in this study. 157 and 98 patients had elevated NLR and PLR respectively (NLR >3.3 and PLR >150). The median overall survival time (OS) and disease-free survival (DFS) was 34.1 and 19.2 months respectively. Multivariate analysis found PLR >150 (P = 0.018, HR 1.426, 95%CI 1.063-1.912) accompanied by male, lymphatic metastases, tumour size more than 3 cm, tumour located at middle segment and poor differentiation were associated with significantly worse DFS. Meanwhile, gender, lymphatic metastases, tumour location and differentiation along with PLR >150 (P = 0.003, HR 1.595, 95% CI 1.172-2.170) and NLR>3.3 (P = 0.039, HR 1.367, 95% CI 1.015-1.840) were all independent prognostic factors for OS. Preoperative NLR and PLR might be used as predictive factors in patients with ESCC. For DFS, elevated PLR compared to NLR may have an advantage to indicate poor prognosis.

High expression of long non-coding RNA ATB indicates a poor prognosis and regulates cell proliferation and metastasis in non-small cell lung cancer.

BACKGROUND AND AIM: Long non-coding RNAs (lncRNAs) have been demonstrated to act as a critical regulator in the processes of tumor biology. In this study, whether lncRNA-ATB is a potential indicator for non-small cell lung cancer (NSCLC) was investigated and its biological function in NSCLC was also determined. METHODS: The expression levels of lncRNA-ATB in NSCLC tissues and cell lines were measured. A549 cell line was explored to investigate the functions of lncRNA-ATB in NSCLC. RESULTS: Real-time PCR results showed that lncRNA-ATB expression was up-regulated in both in NSCLC tissues and cell lines. High lncRNA-ATB expression in tumor tissue was associated with larger tumor size, lymph node metastasis, and distant metastasis in patients with NSCLC, respectively. In addition, the patients with high expression of lncRNA-ATB presented a lower survival probability. In vitro experiments showed that down-regulation of lncRNA-ATB promoted the cell apoptosis, whereas inhibited the cell viability, cell migration, and cell invasion. CONCLUSION: High expression of lncRNA-ATB indicated a poor prognosis and led to the cell proliferation and metastasis in NSCLC.