Lacticaseibacillus rhamnosus GG Improves Periodontal Bone Repair via Gut-Blood Axis in Hyperlipidemia.

Periodontal bone regeneration remains a clinical challenge, and hyperlipidemia can aggravate alveolar bone resorption. Probiotics have recently been reported to improve bone mass. We aimed to determine the role of Lacticaseibacillus rhamnosus GG (LGG) in periodontal bone regeneration improvement within the context of periodontitis with hyperlipidemia. A Sprague Dawley rat model for periodontitis, hyperlipidemia, and periodontal fenestration defect was constructed (n = 36) and administered LGG gavage for 6 wk (the rats were subsequently sacrificed). Fecal microbiota from donor rats 3 wk after LGG gavage was transplanted into recipient rats to evaluate the role of LGG-modulated gut microbiota in periodontal bone regeneration. Regenerated bone mass was detected using micro-computerized tomography and hematoxylin and eosin stain. Gut microbiota was analyzed using 16S ribosomal RNA sequencing. Serum metabolites were detected by liquid chromatography-mass spectrometry (6 wk after LGG gavage). The pro-osteogenic effects of screened serum metabolite were verified in vitro on bone marrow mesenchymal stem cells (BMMSCs). We found that the bone mineral density, bone volume (BV), trabecular bone volume fraction (BV/TV), and trabecular thickness of the regenerated periodontal bone increased after LGG gavage (P < 0.05) but had little effect on oral flora. After LGG gavage, Staphylococcus, Corynebacterium, and Collinsella in the gut of donors were significantly changed, and these differences were maintained in recipients, who also showed increased trabecular thickness of the regenerated periodontal bone (P < 0.05). These key genera were correlated with BV/TV and BV (P < 0.05). In addition, LGG gavage significantly regulated bone-related blood metabolites, of which selenomethionine promoted BMMSC osteogenesis. Notably, selenomethionine was associated with key gut genera (P < 0.05). Collectively, LGG improved periodontal bone regeneration in the context of periodontitis with hyperlipidemia by modulating gut microbiota and increasing pro-osteogenic metabolites in the blood. These results reveal new insights into the use of probiotics to promote periodontal bone regeneration via the gut-blood-bone axis.

[Establishment and validation of a risk predictive model of preoperative drug-induced limitation of pupil dilation in type 2 diabetes mellitus patients with concomitant cataract].

Objective: To establish and validate a risk predictive model of preoperative drug-induced limitation of pupil dilation (PD) in type 2 diabetes mellitus (T2DM) patients with concomitant cataract. Methods: A cross-sectional study was performed, in which 376 T2DM patients with concomitant cataract who received cataract operation in the Second Affiliated Hospital of Zhejiang University School of Medicine from October 2022 to March 2023 were randomly selected as the study subjects. Of the 376 patients, 268 who were admitted to the hospital from October to December 2022 served as the modeling group, and were divided into PD limited group (n=187) and PD unlimited group (n=81) based on whether they had drug-induced limitation of PD. Logistic regression was used to establish a risk predictive model, R software was used to draw the nomogram, Hosmer-Lemeshow test was utilized to judge the model's goodness of fit, and receiver operating characteristic (ROC) curve was adopted to validate the predicting efficacy of the model. Another 108 T2DM patients who received cataract operation in the same hospital from January to March 2023 served as the validation group, and Hosmer-Lemeshow test and ROC curve were used for the external validation of the model. Results: In the modeling group (n=268), there were 124 males and 144 females, with the mean age of (66.6±6.8) years, while in the validation group (n=108), there were 51 males and 57 females, with the mean age of (64.9±9.1) years. The incidence of preoperative drug-induced limitation of PD was 69.8% (187/268) in T2DM patients with concomitant cataract. T2DM disease course (OR=1.134, 95%CI: 1.074-1.198, P<0.001), body mass index (BMI) (OR=0.863, 95%CI: 0.767-0.972, P=0.015), glycohemoglobin (HbA1c) level (OR=1.397, 95%CI: 1.055-1.849, P=0.019) and baseline pupil dimeter (OR=0.089, 95%CI: 0.045-0.179, P<0.001) were the risk factors of drug-induced limitation of PD. Hosmer-Lemeshow test showed χ2=6.231 and P=0.621, the area under curve (AUC) of ROC curve was 0.897 (95%CI: 0.857-0.937, P<0.001), and when the Youden index was the maximum (0.655), the model's sensitivity and specificity was 0.877 and 0.778, respectively. The external validation results demonstrated that the AUC of ROC curve was 0.928 (95%CI: 0.875-0.981, P<0.001), the maximum Youden index was 0.761, the sensitivity was 0.932, the specificity was 0.829, and the overall accuracy was 89.8%. Conclusion: The risk predictive model established in the current study can provide reference for the clinical assessment of the risk of preoperative drug-induced limitation of PD in T2DM patients with concomitant cataract.

[Structural characteristics of lower respiratory tract microflora in patients with pneumoconiosis].

Objective: To explore the composition of bacteria in lower respiratory tract of patients with pneumoconiosis and dust exposure, and to compare and analyze the difference and correlation between them. Methods: From May 2020 to January 2021, a prospective multicenter cross-sectional study was conducted to select patients with pneumoconiosis who underwent bronchoalveolar lavage treatment at the Respiratory and Critical Care Medical Department of the 920th Hospital of the Joint Support Force and the Respiratory Department of Tongren Hospital in Kunming, as well as the population of dust recipients. A total of 24 patients with pneumoconiosis (pneumoconiosis group) were included, and 16 dust exposed individuals (dust exposed group) were used as controls. Two groups of patients' alveolar lavage fluid were collected. The 16SrRNA gene V3-V4 sequencing technology and bioinformatics analysis platform were used to measure and analyze the differences in microbial structure composition and associations between bacterial communities. Results: Compared with the dust exposed group, the top 5 bacterial phyla in the alveolar lavage fluid level of patients with pneumoconiosis were the same, followed by Proteobacteria, Firmicutes, Bacteroidetes, Fusobacteria, and Actinobacteria. Compared with the dust exposure group, the pneumoconiosis group patients belong to the top 5 genera of horizontal flora abundance, which are different. The dust exposure group is respectively: Pseudomonas, Proctor, Streptococcus, Achromobacter, and Neisseria. The pneumoconiosis group is respectively: Pseudomonas, Achromobacter, Streptococcus, Ralstonia, and Proctor. The Alpha diversity analysis results showed that compared with the dust exposed group, the level of bacterial diversity in the pneumoconiosis group was difference (P<0.05), and there was no statistically significant difference in bacterial evenness (P>0.05) ; Beta diversity showed differences in microbial community structure between the two groups (P<0.05 ). Single factor microbial association network analysis showed that there was a high correlation between Firmicutes and Bacteroidetes in the pneumoconiosis and dust exposed groups and other species, showing a positive correlation; The correlation between Proteobacteria and other species is high, showing a negative correlation. Conclusion: The structure and relative abundance of bacteria in lower respiratory tract were different between patients with pneumoconiosis and dust exposure, and the diversity of bacteria in lower respiratory tract increased in patients with pneumoconiosis, which may be related to disease status.

[Analysis of Dimethyltryptamine in Captured Suspicious Substances：Three Case Reports].

ABSTRACT: Abstract.

An algorithm to simulate nonstationary and non-Gaussian stochastic processes.

We proposed a new iterative power and amplitude correction (IPAC) algorithm to simulate nonstationary and non-Gaussian processes. The proposed algorithm is rooted in the concept of defining the stochastic processes in the transform domain, which is elaborated and extend. The algorithm extends the iterative amplitude adjusted Fourier transform algorithm for generating surrogate and the spectral correction algorithm for simulating stationary non-Gaussian process. The IPAC algorithm can be used with different popular transforms, such as the Fourier transform, S-transform, and continuous wavelet transforms. The targets for the simulation are the marginal probability distribution function of the process and the power spectral density function of the process that is defined based on the variables in the transform domain for the adopted transform. The algorithm is versatile and efficient. Its application is illustrated using several numerical examples.

Negative associations of morning serum cortisol levels with obesity: the Henan rural cohort study.

AIMS: To evaluate the associations of morning serum cortisol levels with obesity defined by different indices in Chinese rural populations. MATERIALS AND METHODS: A cross-sectional study was performed including 6198 participants (2566 males and 3632 females). Serum cortisol was collected in morning and quantified by liquid chromatography-tandem mass spectrometry. Obesity was defined by body mass index (BMI), body fat percentage (BFP), waist-to-height ratio (WHtR), waist circumference (WC), visceral fat index (VFI) and waist-to-hip ratio (WHR). Both multivariable liner regression, logistic regression and restrictive cubic splines models were used to estimate the gender-specific relationships between cortisol levels and obesity defined by different indices, respectively. RESULTS: After adjusting for potential confounders, serum cortisol was negatively associated with different obesity measures, except obese females defined by BFP (for instance, overall obesity defined by BMI, Quartile 4 vs. Quartile 1, odds ratio (OR) = 0.25, 95% confidence interval (CI):0.15, 0.41 in males, and OR = 0.58, 95% CI: 0.42,0.80 in females, central obesity defined by WC, OR = 0.52, 95% CI:0.39,0.69 in males and OR = 0.63, 95% CI:0.51,0.77 in females). Similarly, restrictive cubic splines showed the nonlinear relationship between high levels of cortisol and different obesity indices. Furthermore, ROC curve analysis indicated that cortisol could improve the discrimination of model with common biomarkers. CONCLUSION: Morning serum cortisol were negatively related to obesity defined by different indices in Chinese rural populations. In addition, cortisol could be as a biomarker for prediction of obesity in males.

Retrospective Analysis of 61 Cases of Children Died of Viral Pneumonia.

ABSTRACT: Objective To retrospectively analyze the forensic pathological postmortem examination and clinical data of children who died of viral pneumonia in identification of cause of death cases and to discuss the clinical characteristics and pathological features of viral pneumonia in children, in order to provide reference to pathological diagnosis of viral pneumonia in children caused by 2019 novel coronavirus （2019-nCoV） infection. Methods Postmortem examination data from 61 cases of children whose causes of death were identified as viral pneumonia in recent years were collected from the Center of Forensic Identification, Southern Medical University. The gender, age, clinical symptoms and pathological features were comparatively analyzed. Results Among the 61 cases of children who died of viral pneumonia, most were within 2 years old （83.61%）, and a large proportion died within 2 weeks after the onset of the disease （91.80%）. Gross changes in postmortem examination included respiratory mucosal hyperemia, pleural effusion, pulmonary swelling, variegated pulmonary pleura and serosa, as well as focal pulmonary hemorrhage and pulmonary edema. A large proportion of sick children had enlarged mesenteric lymph nodes （83.61%） and thymic dysplasia （21.31%）. Histopathological changes included edema of alveoli and interstitial substance, pneumorrhagia，shedding of alveolar epithelial cells, serous and （or） fibrous exudation in the alveoli, formation of viral inclusions, formation of transparent membranes, infiltration of inflammatory cells that mainly consisted of macrophages and lymphocytes in interstitial substance and alveoli. Viral infections often affected the heart and gastrointestinal tract. Conclusion The clinical symptoms of children with viral pneumonia are difficult to notice, and because the immune systems of children are not fully developed and they have poor immunity, they can easily become severely ill and even die. Analyzing the forensic autopsies and the histopathological characteristics could provide reference for pathological diagnosis of viral pneumonia.

[Epidemiological analysis on a family cluster of COVID-19].

Objective: To understand the possible transmission route of a family cluster of COVID-19 in Zhengzhou and the potential infectivity of COVID-19 in incubation period, and provide scientific evidence for the timely control of infectious source and curb the spread of the epidemic. Methods: Epidemiological investigation was conducted for a family cluster of COVID-19 (8 cases) with descriptive epidemiological method, and respiratory tract samples of the cases were collected for the nucleic acid detection of virus by RT-PCR. Results: Two primary cases, which occurred on 31 January and 1 February, 2020, respectively, had a common exposure history in Wuhan. The other six family members had onsets on 30 January, 31 January, 1 February (three cases) and 3 February, 2020. Conclusions: In this family cluster of COVID-19, six family members were infected through common family exposure to the 2 primary cases. Five secondary cases had onsets earlier than or on the same day as the primary cases, indicating that COVID-19 is contagious in incubation period, and the home isolation in the early phase of the epidemic might lead to the risk of family cluster of COVID-19.

Association of obesity with bone mineral density and osteoporosis in adults: a systematic review and meta-analysis.

OBJECTIVES: Results from previous studies investigating the association of obesity with bone mineral density (BMD) and osteoporosis (OP) are inconsistent. The purpose of the present study was to examine whether obesity is associated with BMD and the risk of OP. STUDY DESIGN: This is a meta-analysis. METHODS: A computerised literature search was conducted using the PubMed, Web of Science, China National Knowledge Internet (CNKI) (Chinese) and Wanfang (Chinese) databases for relevant articles published in English or Chinese before the end of December 2017. Means with standard deviations and odds ratios (ORs) with corresponding 95% confidence intervals (95% CIs) were computed to estimate the association between obesity and BMD and the risk of OP by using a random-effects model. In addition, a heterogeneity test and sensitivity analysis were conducted. RESULTS: Adults with obesity had higher BMD than healthy-weight subjects in both the lumbar spine and femoral neck measurement sites. Obesity was positively related to BMD in the two sites, with merged weighted mean difference (WMD) of 0.07 g/cm2 in the lumbar spine (95% CI: 0.045, 0.095; P < 0.001; Pheterogeneity <0.001; I2 = 89.1%) and WMD of 0.087 g/cm2 in the femoral neck (95% CI: 0.063, 0.112; P < 0.001; Pheterogeneity <0.001; I2 = 92.8%). General obesity was negatively related to femoral neck OP, indicating that obesity is a protective factor for OP, with a merged OR of 0.169 (95% CI: 0.132, 0.216; P < 0.001; Pheterogeneity = 0.716; I2 = 0.0%). CONCLUSIONS: Adults with obesity had significantly higher BMD than healthy-weight adults. Obesity was positively associated with BMD and negatively correlated with OP.

Lipid profiles as potential mediators linking body mass index to osteoporosis among Chinese adults: the Henan Rural Cohort Study.

This study is to examine the relationship between body mass index (BMI) and osteoporosis in rural areas of China, and then explore whether this association was mediated by lipid profiles. Results showed that with the increasing of BMI, the risk of osteoporosis was reduced, and lipid profiles mediate this association. INTRODUCTION: To examine the relationship between BMI and osteoporosis, and explore whether this association was mediated by lipid profiles. METHODS: A total of 8272 participants (18-79 years) were enrolled from the Henan Rural Cohort Study. The bone mineral density of the calcaneus was measured using an ultrasonic bone density apparatus. Logistic regression and restricted cubic splines were used to evaluate the odds ratio (OR) and 95% confidence intervals (95% CI). Mediation analysis using bootstrap was performed to examine the contribution of lipid profiles to BMI-related osteoporosis. RESULTS: The crude and age-standardized prevalence of osteoporosis were 15.93% and 11.77%, respectively. The mean BMIs were 24.12 kg/m2 for participants with osteoporosis and 25.06 kg/m2 for non-osteoporosis participants (P < 0.001). After adjusting for potential confounders, subjects with obesity had a lower OR of osteoporosis (0.493 [95% CI: 0.405-0.600], Ptrend < 0.001) compared with normal-weight individuals. Mediation analysis showed that lipid profile partly mediated the relationship between BMI and osteoporosis with indirect effect OR (95% CI) of 0.985 (0.978-0.992), and the proportion explained of BMI was 15.48% for lipid profile. CONCLUSION: With the increasing of BMI, the risk of osteoporosis was reduced in the Chinese adult population, and lipid profiles may be a potential mediator linking reduced risk of osteoporosis. Elucidating the underlying mechanisms will facilitate developing feasible preventive and therapeutic measures for osteoporosis. Chinese clinical trial register: ChiCTR-OOC-15006699.

Retrospective Analysis of 291 Cases of Medical Malpractice Involving Death.

ABSTRACT: Objective To study the medical malpractice cases involving death, and discuss the identification ideas and methods of medical malpractice cases. Methods A total of 291 medical malpractice cases involving death accepted and settled from January 2012 to December 2017 at the Judicial Appraisal Center of Southern Medical University were collected. Based on the age, gender, hospital level, clinical department, whether or not autopsy was performed, cause of death, cause of medical mistakes, causality and causative potency of the appraised person, statistical analysis was made. Results There were more males than females in medical malpractice cases involving death. Mostly young adults or children were involved in these cases. The number of cases involving tertiary hospitals was the highest; among the clinical departments, the internal medicine department had the largest number of cases, followed by surgery, obstetrics and gynecology, pediatrics, etc. Autopsy rate has a trend of increasing year by year. Most patients die from the natural outcomes of their disease or ineffective treatment. Most hospitals have certain medical mistakes, and have an indirect correlation with the patient's death, mainly slight factors. Conclusion Judicial appraisal of medical malpractice should follow the principle of "one-effect and multi-cause", and comprehensively consider various factors such as, the diseases and constitution of the patient, natural outcomes of the diseases, the current medical technology and the level of diagnosis and treatment of the hospital, etc.

Expression of microRNA-122 and microRNA-22 in HBV-related liver cancer and the correlation with clinical features.

OBJECTIVE: MicroRNAs (miR) participate in cell proliferation, apoptosis and transformation, as they can regulate gene expression and intracellular signal transduction for various physiological processes. MiR-122 and miR-22 are known to be related with occurrence and progression of hepatitis B virus (HBV)-related hepatocellular cancer (HCC). This study recruited HBV-related HCC patients, whose expression levels of miR-122 and miR-22 were determined to analyze the correlation with clinical and pathological indexes. PATIENTS AND METHODS: HBV-related HCC patients were enrolled, in parallel with patients suffering from benign liver disease and non-HBV-related HCC. Real-time PCR was employed to measure miR-122 and miR-22 expression levels. RESULTS: The relative expression levels of miR-122 and miR-22 in HBV-related HCC patients were 1.26 ± 2.73 and 5.49 ± 3.91, respectively, which were significantly lower than that in benign liver disease or non-HBV-related HCC patients (p < 0.05). No significant difference of serum miR-122 or miR-22 levels was found between benign liver disease and non-HBV-related HCC patients (p > 0.05). The miR-122 and miR-22 levels were negatively correlated with tumor size, lymph node metastasis, TNM stage, pathological type, differentiation grade, liver cirrhosis, AFP and HBV DNA, all of which were independent risk factors (p < 0.05). CONCLUSIONS: MiR-122 and miR-22 were downregulated in HBV-related HCC patients, and were related with tumor size, lymph node metastasis, TNM stage, pathological type, differentiation grade, liver cirrhosis, AFP and HBV DNA.

Associations of serotonin receptor gene HTR3A, HTR3B, and HTR3A haplotypes with bipolar disorder in Chinese patients.

Single nucleotide polymorphisms (SNPs) in HTR3A and HTR3B have been reported to be associated with bipolar disorder in European and Japanese populations. We explored the roles of 21 tag SNPs in HTR3A and HTR3B in susceptibility to bipolar disorder in a Chinese cohort. Twenty-one Tag SNPs were genotyped in a study consisting of 130 patients with bipolar disorder, who visited Shandong Mental Health Center between June 2013 and May 2014, and 109 healthy individuals as controls. All of the tag SNPs were genotyped using Sequenom MassArray matrix-assisted laser desorption/ionization time of flight spectrometry. Plink 1.07, Haploview 4.2, and SPSS 20.0 were used for the analysis of the genotypes and the associations of the haplotypes with bipolar disorder. Association analyses of tag SNPs detected significant associations with the A allele in HTR3A rs1176719 (P = 0.030) and the C allele in HTR3A rs1176713 (P = 0.048). Haplotype-based association analyses indicated a statistically significant (P = 0.035) five-SNP haplotype (rs1062613:C, rs11604247:C, rs1176722:G, rs2276302:A, rs1176719:G) of linkage disequilibrium in block 3. Analysis of our small Chinese sample revealed a significant association of HTR3A with bipolar disorder, but yielded no evidence of an association between HTR3B and bipolar disorder. Furthermore, evidence for an association was found for a haplotype of HTR3A. Studies with larger Chinese samples are needed to verify our findings.

[In vitro study of biological characteristics of new retrograde filling materials iRoot].

OBJECTIVE: To study the biological characteristics of new retrograde filling materials iRoot BP plus and iRoot FS. METHODS: (1) The roots were cut into 3 mm in length, and the root canals were prepared to 1 mm in diameter, followed by being filled with iRoot BP plus, iRoot FS, or mineral trioxide aggregate (MTA). The specimens were immersed in simulated body fluid (SBF). The ability of mineralization in vitro was detected through three studies. First, the mineralization of specimens was analyzed through scanning electron microscope observations and energy dispersive X-ray spectrometer. Then, the pH of SBF was monitored using pH meter. (2) The extracts were gained by immersing blocks of iRoot BP plus, iRoot FS, and MTA (8 mm diameter and 2 mm height) into dulbecco's modified eagle medium (DMEM). The effects of the extracts on proliferation of MG63 cells were detected through MTT assay. The gene expression level of alkaline phosphatase (ALP) was analyzed by quantitative real-time PCR, and the expression of ALP activity was observed by ALP activity staining. RESULTS: (1) The formation of minerals could be observed on the surfaces of iRoot BP plus, iRoot FS, and MTA at the end of 24 h, and there were more amounts of apatite aggregated after 14 days. The values of Ca/P ratios of apatites were 1.43, 1.39, and 1.51, respectively. (2) The pH of SBF could be raised to 8.09±0.07, 7.91±0.06, and 8.11±0.06, respectively, significantly higher than the blank. (3) The extracts of iRoot BP plus, iRoot FS, and MTA of dilutions of 1:5 and 1:10 presented no effect of proliferation of MG63 cells. (4) iRoot BP plus and iRoot FS could significantly up-regulated the levels of ALP messenger RNA expression, while there was no obvious difference in ALP staining among the iRoot BP plus, iRoot FS, MTA, and the blank. CONCLUSION: The present study shows that iRoot has displayed good mineralization capability in vitro and capability to promote differentiation and mineralization of MG63 cells, inferring that iRoot may have good bioactivity.

Tricyclic Neovibsanin Scaffold (TCNS) as an Effective Antidepressant Agent.

Neovibsanin type natural products were found to display neurite outgrowth activity in PC12 cells. This suggests that such type of compounds could be promising candidates for the development of novel therapeutic agents to treat neurological diseases. In the present study rats after chronic mild stress (CMS) were treated with tricyclic neovibsanin scaffold (TCNS) to study its effect on depression. The results revealed that 15 mg/kg doses of TCNS reduced the duration of immobility in CMS model of depression. It led to a significant increase in neurite outgrowths which increased the synaptic and structural plasticity of neurons. Treatment with TCNS decreased the levels of MAO-A and caspase-3 expression both of which were found to be higher in CMS. TCNS also led to an increase in expression of heat shock protein 70 (Hsp70) in the hippocampus. These findings suggest that TCNS possesses antidepressant activity in CMS model of depression. Therefore the relief in depression by TCNS may be due to suppression of MAO-A expression and the apoptosis cascade by increased expression of Hsp70.

Increased extracellular dopamine and 5-hydroxytryptamine levels contribute to enhanced subthalamic nucleus neural activity during exhausting exercise.

The purpose of the study was to explore the mechanism underlying the enhanced subthalamic nucleus (STN) neural activity during exhausting exercise from the perspective of monoamine neurotransmitters and changes of their corresponding receptors. Rats were randomly divided into microdialysis and immunohistochemistry study groups. For microdialysis study, extracellular fluid of the STN was continuously collected with a microdialysis probe before, during and 90 min after one bout of exhausting exercise. Dopamine (DA) and 5-hydroxytryptamine (5-HT) levels were subsequently detected with high-performance liquid chromatography (HPLC). For immunohistochemistry study, the expression of DRD2 and HT2C receptors in the STN, before, immediately after and 90 min after exhaustion was detected through immunohistochemistry technique. Microdialysis study results showed that the extracellular DA and 5-HT neurotransmitters increased significantly throughout the procedure of exhausting exercise and the recovery period (P<0.05 or P<0.01). Immunohistochemistry study results showed that the expression levels of DRD2 and HT2C in the rat STN immediately after exhausting exercise and at the time point of 90 min after exhaustion were both higher than those of the rest condition, but the difference was not significant (P>0.05). Our results suggest that the increased extracellular DA and 5-HT in the STN might be one important factor leading to the enhanced STN neural activity and the development of fatigue during exhausting exercise. This study may essentially offer useful evidence for better understanding of the mechanism of the central type of exercise-induced fatigue.

Glans-reconstruction with preputial flap is superior to primary closure for post-surgical restoration of male sexual function in glans-preserving surgery.

We conducted this study to investigate whether glans-reconstruction with preputial flap would be more helpful for post-operative restoration of patients' sexual potency than primary closure. From 2007 to 2013 at four centres, 46 cases reconstructed with preputial flap and 59 with primary closure were selected for the investigation from 142 consecutive cases of superficial glans cancer, with the largest diameter of lesion ≤2.5 cm, who undergoing glans-preserving surgery (GPS). Subjective evaluation for patients' sexual performance was investigated using the International Index of Erectile Function-15. Objective evaluation was carried out by the Audio Visual Sexual Stimulation test with RigiScan-Plus. The degree of satisfaction for penile appearance and patients' confidence and partners' acceptability for intercourse were evaluated by 5-point scales. Patients with preputial flap reconstruction showed significant better performance in three domains (orgasmic function, intercourse satisfaction and overall satisfaction, all p < 0.05) and significantly higher ratios of appearance satisfaction (78.3% vs. 57.6%, p = 0.026) and intercourse confidence (69.6% vs. 49.2%, p = 0.035) compared with those undergoing primary closure at post-operative month 6 ends. Their sexual partners in the preputial flap group also exhibited significantly higher ratios of appearance satisfaction than in the primary closure group (67.4% vs. 42.4%, p = 0.011). Reconstruction with preputial flap contributes to a more acceptable cosmetic appearance of the penis and minimizes post-operative negative psychological impediments. Patients can benefit more from reconstruction with preputial flap than primary closure. Glans-reconstruction with preputial flap should be considered the primary reconstruction technique in GPS.

ESAT-6- and CFP-10-specific Th1, Th22 and Th17 cells in tuberculous pleurisy may contribute to the local immune response against Mycobacterium tuberculosis infection.

Th1 cell-mediated adaptive immune response is very important but may not be sufficient to control Mycobacterium tuberculosis (M. tuberculosis) infection. The roles of the various T cell subsets and cytokines in the inflammatory processes are not clearly elucidated. We investigated whether Th1, Th22 and Th17 cells mediated cellular immunity at the local site of M. tuberculosis infection in patients with tuberculous pleurisy (TBP). The results showed that the cytokines IFN-γ and IL-22 but not IL-17 were elevated in tubercular pleural fluid. Following stimulation with immune-dominant peptides of early secreted antigenic target-6 (ESAT-6), culture filtrate protein-10 (CFP-10) or Bacille Calmette-Guerin, pleural fluid mononuclear cells expressed high levels of cytokines IFN-γ, IL-22 and IL-17 as revealed by mRNA and protein measurements. In addition, we showed that cytokines IFN-γ, IL-22 and IL-17 were produced in M. tuberculosis-specific immune response by distinct subsets of CD4+ T cells with the phenotype of CD45RA-CD62L-CCR7+CD27+ . Our results demonstrated for the first time that ESAT-6- and CFP-10-specific Th1, Th22 and Th17 cells existed in the patients with TBP and might play an essential role against M. tuberculosis infection. The findings of this study raised the possibility of unravelling the critical targets for therapeutic intervention in chronic inflammatory diseases such as TBP.

Efficient fluorescence-based imaging methods for quantitating infectivity and oncolytic efficacy of Newcastle disease virus.

A high throughput quantitation protocol is desired to determine the replication of various recombinant oncolytic viruses in vitro. Plaque assay is the classic method for viral infectivity quantitation but is laborious and time consuming; moreover it does not report the oncolytic efficacy of a virus. In this paper, three new imaging methods for quantitating viral infectivity are derived and evaluated: fluorescence intensity, infection counts, and infection degree. Infection of oncolytic Newcastle disease virus in human tumor and normal cells was followed over a time course by plaque assay and the imaging methods. For the latter, brightfield and green channel images were acquired at various fixed locations in the cell culture, and later analyzed. One of the imaging methods was found to be highly correlated with viral titer; the other methods are complementary to plaque assay and provide additional information like oncolytic efficacy, syncytium formation etc. The new methods significantly reduce the time and material costs required by plaque assay, and provide an efficient system for quantitating and characterizing infectivity and efficacy of oncolytic viruses.

MicroRNA-25 functions in regulation of pigmentation by targeting the transcription factor MITF in Alpaca (Lama pacos) skin melanocytes.

Although the influence of endocrine factors is well established, the molecular and cellular mechanisms controlling coat color are not completely understood. A major mechanism for post-transcriptional regulation of gene expression is through the action of microRNAs (miRNAs), which anneal to the 3' untranslated region of mRNAs in a sequence-specific fashion and either block translation or promote transcript degradation. In this study, we investigated the expression of miRNAs in the skin of alpacas with brown vs white coat color using a microarray screen; identified potential mRNA targets for identified miRNAs among coat color genes; and subsequently determined the ability of a specific, differentially expressed miRNA (miR-25) to suppress expression of micropthalmia-associated transcription factor (MITF), a predicted miR-25 target gene that regulates genes linked to coat color. Expression of 10 different miRNA species in the skin of alpacas with brown vs white coat color was identified from microarray screens. Of the 10 alpaca skin miRNAs identified, predicted binding sites in the 3' untranslated region of RNAs encoding for known genes linked to coat color were primarily for miR-25, but sites were also identified for miR-129 and miR-377. Potential miR-25 binding sites were present in transcripts encoding for 11 coat color genes, including MITF. An inverse relationship between transcript abundance for MITF and miR-25 was observed in skin samples collected from alpacas with white vs brown coat color. Furthermore, overexpression of miR-25 in cultured melanocytes reduced MITF mRNA and protein abundance and corresponding mRNA abundance for the MITF-regulated enzymes tyrosinase and tyrosinase-related protein 1. Results support a novel functional role for miRNA-25 in the regulation of gene expression linked to coat color.