Characterization of the protein expression and localization of hnRNP family members during murine spermatogenesis.

Mammalian testis exhibits remarkably high transcriptome complexity, and spermatogenesis undergoes two periods of transcriptional cessation. These make the RNA-binding proteins (RBPs) the utmost importance during male germ cell development. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are a large family of RBPs implicated in many steps of RNA processing; however, their roles in spermatogenesis are largely unknown. Here, we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis. Furthermore, we found that most of the detected hnRNP proteins (hnRNPD, hnRNPK, hnRNPQ, hnRNPU, and hnRNPUL1) display the highest signals in the nuclei of pachytene spermatocytes, round spermatids, and Sertoli cells, whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids. The expression of these hnRNP proteins showed both similarities and specificity, suggesting their diverse roles in spermatogenesis.

Cryoablation treatment of oral and maxillofacial tumors: Five case reports.

Oral and maxillofacial anatomy and function are important and complex. They are involved in facial expressions, chewing, language, breathing, and other functions. It is therefore important to choose the optimal treatment plan for oral and maxillofacial tumors. For patients with who cannot tolerate surgery or who refuse surgery or radiotherapy can be treated with cryoablation. Cryoablation can maintain local tissue integrity and organ function and protect facial integrity. It is a repeatable treatment that, if necessary, can be followed by traditional antineoplastic therapies. This study introduces five cases with severe basic diseases who cannot tolerate or have refused surgery or radiotherapy. The patients were diagnosed as having oral and maxillofacial tumors. These patients experienced painful local swelling or breaking of the tumor. All patients received cryoablation combined with other treatments. Local control of the tumors and improved function and quality of life were achieved. In clinical work, for patients with severe basic diseases who cannot tolerate or refuse surgery or radiotherapy, cryoablation has unique advantages, and this approach is expected to become a widely used treatment for oral and maxillofacial tumors.

Circular RNA expression alteration and bioinformatics analysis in patients with acute cerebral infarction injury.

In recent years, a steady increase has been detected in the incidence of acute cerebral infarction (ACI). ACI is caused by blood flow disruption, leading to high disability and mortality rates. Understanding the underlying molecular mechanisms is critical toward developing effective therapeutic approaches. Circular RNAs (circRNAs) are an important class of non-coding RNAs, which have been implicated in several molecular pathways, and their dysregulation has been described in several disease conditions. Here, we set out to explore the possible regulatory role of circRNAs in ischemic stroke and study their molecular function in disease. First, we applied high-throughput sequencing techniques to identify the differential changes of plasma circRNAs expression in patients with acute cerebral infarction. Next, we used GO and KEGG pathway analysis to predict the function of differentially expressed circRNAs. Moreover, we have assessed the possible interaction between the identified differentially expressed circRNAs and miRNAs. Finally, we have selected and validated five downregulated circRNAs by RT-qPCR. Together, the results of this study provide evidence that circRNAs are potential biomarkers for early diagnosis of cerebral infarction and have to be considered as targets for drug treatment.

Proteome sequencing and analysis of Ophiocordyceps sinensis at different culture periods.

BACKGROUND: Ophiocordyceps sinensis is an important traditional Chinese medicine for its comprehensive active ingredients, such as cordycepin, cordycepic acid, and Cordyceps polysaccharide. O. sinensis zjut, a special strain isolated from O. sinensis, has similar pharmacological functions to wild O. sinensis. Currently, O. sinensis with artificial cultivation has been widely studied, but systematic fundamental research at protein levels has not been determined. RESULTS: Proteomes of O. sinensis zjut at different culture periods (growth period, 3rd day; pre-stable period, 6th day; and stable period, 9th day) were relatively quantified by relative isotope markers and absolute quantitative technology. In total, 4005 proteins were obtained and further annotated with Gene Ontology, Kyoto Encyclopedia of Genes and Genomes database. Based on the result of the annotations, metabolic pathways of active ingredients, amino acids and fatty acid were constructed, and the related enzymes were exhibited. Subsequently, comparative proteomics of O. sinensis zjut identified the differentially expressed proteins (DEPs) by growth in different culture periods, to find the important proteins involved in metabolic pathways of active ingredients. 605 DEPs between 6d-VS-3d, 1188 DEPs between 9d-VS-3d, and 428 DEPs between 9d-VS-6d were obtained, respectively. CONCLUSION: This work provided scientific basis to study protein profile and comparison of protein expression levels of O. sinensis zjut, and it will be helpful for metabolic engineering works to active ingredients for exploration, application and improvement of this fungus.

Genome sequencing and analysis of fungus Hirsutella sinensis isolated from Ophiocordyceps sinensis.

Ophiocordyceps sinensis has been used as a traditional medicine or healthy food in China for thousands of years. Hirsutella sinensis was reported as the only correct anamorph of O. sinensis. It is reported that the laboratory-grown H. sinensis mycelium has similar clinical efficacy and less associated toxicity compared to the wild O. sinensis. The research of the H. sinensis is becoming more and more important and urgent. To gain deeper insight into the biological and pharmacological mechanisms, we sequenced the genome of H. sinensis. The genome of H. sinensis (102.72 Mb) was obtained for the first time, with > 99% coverage. 10,200 protein-encoding genes were predicted based on the genome sequence. A detailed secondary metabolism analysis and structure verification of the main ingredients were performed, and the biosynthesis pathways of seven ingredients (mannitol, cordycepin, purine nucleotides, pyrimidine nucleotides, unsaturated fatty acid, cordyceps polysaccharide and sphingolipid) were predicted and drawn. Furthermore, infection process and mechanism of H. sinensis were studied and elaborated in this article. The enzymes involved in the infection mechanism were also predicted, cloned and expressed to verify the mechanism. The genes and proteins were predicted and annotated based on the genome sequence. The pathways of several active components in H. sinensis were predicted and key enzymes were confirmed. The work presented here would improve the understanding of the genetic basis of this organism, and contribute to further research, production and application of H. sinensis.

Esculetin enhances the inhibitory effect of 5-Fluorouracil on the proliferation, migration and epithelial-mesenchymal transition of colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is the most common malignant disease worldwide and thus new therapeutic approaches are needed. 5-Fluorouracil (5-FU) remains the most widely used agent to treat colorectal cancer (CRC). However, its clinical efficacy is currently limited by the development of drug resistance. Esculetin (EST), a coumarin, was found to have anti-proliferative and anti-migration activity in cancer. OBJECTIVE: This research aims to evaluated the influence and possible mechanism of EST on the proliferation, migration and epithelial-mesenchymal transition of CRC cell lines. MATERIALS AND METHODS: Human CRC cell lines HT-29, SW480, HCT-116, and Caco-2 were treated with various concentrations of EST (0.2, 2, 20, 200, 2000 µg/ml) or 5-FU (0.1, 1, 10, 100, 1000 µg/ml) for 48 h, and cell viability was determined by the MTT and CCK-8 assay. The motility of HCT-116 cells was detected by scratch assay. Western blot was applied to detect the protein expression. Besides, levels of Wnt3a and VEGF in HCT-116 cell culture medium supernatant were analyzed by ELISA. The anti-tumor effect was detected with HCT-116 subcutaneous tumor bearing tumor model by monitoring the tumor vomume in vivo. Finally, the tumoral expression of VEGF was measured by immunohistochemistry, and the expression of Ki67, PCNA, ß-catenin, c-Myc, Cyclin D1, MMP2 and MMP7 was measured by Western blot analysis. RESULTS: EST inhibited HCT-116 cell proliferation in a dose-dependent manner. Western blot analysis revealed that EST decreased the expression of Ki67, PCNA, N-cadherin, E-cadherin, vimentin, fibronectin, ß-catenin, c-Myc, Cyclin D1, MMP2 and MMP7. Furthermore, EST reduced the release of Wnt3a and VEGF into HCT-116 cells culture medium. After EST treatment, the tumor volume was significant smaller than that of the control group, and the tumoral levels of VEGF were decreased. Moreover, western blot analysis indicated that the expression of Ki67, PCNA, ß-catenin, c-Myc, Cyclin D1, MMP2 and MMP7 were also significantly decreased after treated with EST. In addition, in vitro and in vivo anti-tumor results demonstrated that EST combined with 5-FU could increase the inhibitory effect of 5-FU on HCT-116 cells proliferation, migration and epithelial-mesenchymal transition. CONCLUSIONS: EST enhances the inhibitory effect of 5-FU on the proliferation, migration and epithelial-mesenchymal transition of CRC.

Identification of programmed cell death 1 and its ligand in the testicular tissue of mice.

PROBLEM: This study aims to determine the expression and localization of programmed cell death 1 (PD-1) and programmed cell death 1 ligand 1 (PD-L1) in the testes of mice at different developmental stages. METHOD OF STUDY: By means of RT-qPCR, Western blot and immunofluorescence, the expression and localization of PD-1 and PD-L1 were detected in the testicular tissues of mice at different postnatal times: P7, P14, P21, P28, P35, and adulthood. Meanwhile, the level of soluble PD-L1 (sPD-L1) was evaluated by ELISA in the testicular interstitial fluid (IF) of the adult mice, culture supernatants of TM4 cell lines (Sertoli cells lines), and primary Sertoli cells at P14. RESULTS: Pd-1 mRNA levels were unexpectedly low. From P7 to P21, there was limited PD-1 protein detected while PD-1 was evident at P28 and afterward at significantly higher levels than at P14 and P21 (P < 0.05). Despite being found in the interstitial area at P7, P14, and P21, PD-1 was also detected in the germ cells of the seminiferous tubules after P28. Pd-l1 mRNA exhibited age-related changes, peaking at P21, while PD-L1 protein was constitutively expressed at any stage, specifically localized in the nucleus of Sertoli cells. Moreover, the level of sPD-L1 in IF was significantly higher than that in the culture supernatants of both TM4 and primary Sertoli cells at P14. CONCLUSIONS: PD-1 and PD-L1 were present in the testicular tissue of adult mice. The expression and localization of PD-1 fluctuated with age, and PD-1 was mainly localized to advanced germ cells, suggesting that it may play a role in spermiogenesis. PD-L1 was constitutively expressed in the nucleus of Sertoli cells, which could secrete sPD-L1 into the testicular interstitial space and thus may be involved in testicular immune privilege.

Simple and efficient rat model for studying delayed cerebral ischemia after subarachnoid hemorrhage.

BACKGROUND: Delayed cerebral ischemia (DCI) is a late phase of consequences of subarachnoid hemorrhage (SAH) that causes poor outcome and has become the focus of current research. The aim of this study was to characterize an experimental SAH technique for studying DCI after SAH. NEW METHOD: A double injection SAH rat model with a tiny incision was introduced. At 7 days post-SAH induction, the diameter and luminal cross-sectional area (CSA) of the basilar artery (BA) were measured. In vivo fluorescence microscopy and magnetic resonance perfusion-weighted imaging (MRPWI) were used to evaluate the occurrence of DCI. Normal and sham-operated groups served as References RESULTS: Compared to the sham group, in SAH group, the diameter and CSA of the BA were decreased, and the CBF in the SAH group was also reduced to barely half of the level in the sham group. Moreover, both the proportion and severity of microarterial constrictions were increased significantly in the SAH group when compared to those in the sham group. COMPARISON WITH EXISTING METHODS: Complete exposure of the atlanto-occipital membrane is avoided, only a tiny region is exposed to identify the puncture spot. Lower mortality, reduced operative trauma and shorter procedure time are advantages to existing models. Multiple techniques for DCI assessment were used including in vivo microscopy and MRPWI. CONCLUSIONS: The current study demonstrates that our SAH model was successfully established and may serve to help identify a novel target for the treatment of DCI after SAH.

Diagnostic value on the analysis of contrast-enhanced ultrasound and quantitative mode of benign and malignant thyroid nodule and the diagnostic value of quantitative model on it / 中国医学装备

Objective: To study ultrasound contrast enhanced mode of benign and malignant thyroid nodule and the change of related quantitative parameter of time intensity curve(TIC).Methods: Retrospective analysis was applied to analyze the mode that contrast agent went into nodules when 125 thyroid nodules of 112 patients with thyroid nodule received ultrasound contrast,and to analyze enhanced mode and distribution situation of contrast agent.The change of TIC was observed and analyzed.And when contrast agent went into nodule,the quantitative parameters included arrive time(AT),peak intensity(PI),time to peak(TTP)and K value of rate of curve were recorded.Results: 58 nodules were benign and 67 nodules were malignant in the 125 thyroid nodules.The rim of benign nodules appeared circular enhancement(82.7％)after contrast agent went into benign nodule.And the malignant nodules appeared non-circular enhancement(77.6％),and the difference of them was no significant(x2=28.32,P＜0.05).Most of malignant nodules appeared asymmetrical enhancement after contrast agent was infused in them(91.0％),and the difference of ultrasound contrast between benign nodules and malignant nodules was significant(x2=17.56,P＜0.05).When the contrast agent of malignant nodules achieved peak value,its main appearance was low enhancement(83.6％),and the contrast agent of benign nodules achieved peak value,the main appearance was equivalent enhancement(86.2％),and the difference of appearance between malignant nodule and benign nodule was significant(x2=41.65,P＜0.05).The differences of parameters of TIC included AT,PI,TTP and K value between malignant nodules and benign nodules were significant(t=2.964,t=3.021,t=-2.914,t=2.652,P＜0.05).Conclusion: For diagnosis of thyroid nodule that mainly contained solid nodule and cystic solid nodule,ultrasound contrast enhancement and TIC included variously quantitative parameters have higher clinical application value.

Enhancement of Nucleoside Production in Hirsutella sinensis Based on Biosynthetic Pathway Analysis.

To enhance nucleoside production in Hirsutella sinensis, the biosynthetic pathways of purine and pyrimidine nucleosides were constructed and verified. The differential expression analysis showed that purine nucleoside phosphorylase, inosine monophosphate dehydrogenase, and guanosine monophosphate synthase genes involved in purine nucleotide biosynthesis were significantly upregulated 16.56-fold, 8-fold, and 5.43-fold, respectively. Moreover, dihydroorotate dehydrogenase, uridine nucleosidase, uridine/cytidine monophosphate kinase, and inosine triphosphate pyrophosphatase genes participating in pyrimidine nucleoside biosynthesis were upregulated 4.53-fold, 10.63-fold, 4.26-fold, and 5.98-fold, respectively. To enhance the nucleoside production, precursors for synthesis of nucleosides were added based on the analysis of biosynthetic pathways. Uridine and cytidine contents, respectively, reached 5.04 mg/g and 3.54 mg/g when adding 2 mg/mL of ribose, resulting in an increase of 28.6% and 296% compared with the control, respectively. Meanwhile, uridine and cytidine contents, respectively, reached 10.83 mg/g 2.12 mg/g when adding 0.3 mg/mL of uracil, leading to an increase of 176.3% and 137.1%, respectively. This report indicated that fermentation regulation was an effective way to enhance the nucleoside production in H. sinensis based on biosynthetic pathway analysis.

A review of assessment scales of dementia-related knowledge / 中国心理卫生杂志

The public's correct knowledge of dementia is the important prerequisite of the early detection and early intervention.Currently,the dementia-related knowledge is mostly evaluated by using self-assessment scales.Fully understanding the applicable range and advantages and disadvantages of each scale has vital significance in guiding the choice of dementia-related knowledge assessment scale.This article introduces the assessment scales of dementia-related knowledge from scale description,psychometric indexes and scale evaluation,respectively.This article finds that the research on the China' s localization of dementia-related knowledge assessment scale needs further deepening and refining.By this way,the research could truly reflect the different levels of knowledge and deficiencies of the tested object and provide reference to improve the dementia management in China.

Evaluation and related factors of mental health access / 中国心理卫生杂志

Access to mental health services is an important part of health services research,which is of great significance for higher-quality mental healthcare and better health outcomes.This paper introduces the concept and basic evaluation theories of health care access,outline the concept and evaluation of mental health access,synthesizes studies on mental health access,and sorts the related factors into supply-side factors,demand-side factors and social factors.

Cognitive impairment and neural electrophysiological studies in early stage of transgenic Alzheimer's disease mice / 中华行为医学与脑科学杂志

Objective To investigate the early onset of learning and memory function of 4-month-old APP/PS1/Tau Alzheimer' s disease (3×Tg-AD) model mice and explore the pathogenesis of AD in early stage through evaluating neuron excitability and BKCa channel activity in cingulate cortex pyramidal cells.Methods Ten 4-month-old male 3×Tg-AD mice and matched ten wild type (WT) mice.Behavior was tested with the novel object recognition task to observe the ability of learning and memory.Whole-cell patch-clamp recordings were performed to assess the excitability of cingulate cortex pyramidal cells in terms of resting membrane potential and frequencies of spikes evoked by current injection.A train of five pulses of depolarizing currents were injected at 100 Hz to assess the spike width,which was used as an index for BKCa channel activity.Results Compared with the WT group (0.72±0.03),the novel object recognition index significantly decreased in 3 × Tg-AD group (0.55 ± 0.04) (P =0.004).Compared to the WT group((-66.03±0.43) mV),the resting membrane potential in cingulate cortex neurons of 3×Tg-AD group((-62.31±0.54)mV) was significantly depolarized(P=0.000).In contrast to WT group,the action potential firing frequencies evoked by depolarizing current injections were higher in neurons from 3×Tg-AD group(P=0.000),demonstrating that excitability of cingulate cortex neurons was elevated by intracellular Aβ.Spikes were broader in the 3×Tg-AD group than those in the WT group(P＜0.01).Suppression of BKCa channels in cingulate cortex neurons from the 3×Tg-AD group was confirmed on the basis of the spike half-width,since BKCa channels affect the descending phase of spikes.Conclusion Compared to WT mice,4-month-old 3×Tg-AD mice are impaired in learning and memory.The suppression of BKCa channels by intracellular Aβ leads to increase of excitability in cingulate cortex pyramidal cells.

[Effect of shenhuang ointment in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of Qi stagnation blood stasis syndrome: a clinical observation].

OBJECTIVE: To observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS). METHODS: A prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed. RESULTS: Compared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05). CONCLUSIONS: The plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.

Effect of shenhuang ointment in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of Qi stagnation blood stasis syndrome: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS).</p><p><b>METHODS</b>A prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed.</p><p><b>RESULTS</b>Compared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05).</p><p><b>CONCLUSIONS</b>The plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.</p>

The cytoplasmic droplet may be indicative of sperm motility and normal spermiogenesis.

Although the cytoplasm of spermatids is removed at the end of spermiogenesis, a tiny portion is usually retained in the sperm flagellum, which is termed the cytoplasmic droplet (CD) in mammals. CDs are believed to play a role in sperm volume adaptation. However, we have noticed that epididymal spermatozoa that display initial (flagellation in situ) and progressive motility mostly possess CDs, whereas spermatozoa without CDs are rarely motile, suggesting that CDs have a role in motility development during sperm epididymal maturation. In the present study, we analyzed the relationship between the presence or absence of CDs, motility development and positional changes of CDs during sperm epididymal maturation in mice and monkeys. We also examined CDs on spermatozoa of three knockout mouse lines with late spermiogenic defects. Our data suggest that the CD is a normal organelle transiently present exclusively on epididymal spermatozoa, and normal CD morphology and location are associated with normal motility development during epididymal maturation of spermatozoa. Abnormal CD formation, e.g., a complete lack of CDs or ectopic CDs, is indicative of defective spermiogenesis. If CDs are essential for sperm motility development, then CDs may represent an ideal drug target for the development of non-hormonal male contraceptives.