Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
Animals ; Haplorhini ; Axons ; Motor Neurons ; Interneurons ; Macaca ; Dependovirus/genetics* ; Genetic Vectors

By consulting the ancient and moderm literature， this paper makes a textual research on the name， origin， quality evaluation， harvesting and processing of Olibanum， so as to provide a basis for the development of the famous classical formulas containing this medicinal material. According to the herbal textual research， the results showed that Olibanum was first described as a medicinal material by the name of Xunluxiang in Mingyi Bielu（《名医别录》）， until Ruxiang had been used as the correct name since Bencao Shiyi（《本草拾遗》） in Tang dynasty. The main origin was Boswellia carterii from Burseraceae family. The mainly producing areas in ancient description were ancient India and Arabia， while the modern producing areas are Somalia， Ethiopia and the southern Arabian Peninsula. The medicinal part of Olibanum in ancient and modern times is the resin exuded from the bark， which has been mainly harvested in spring and summer. It is concluded that the better Olibanum has light yellow， granular， translucent， no impurities such as sand and bark， sticky powder and aromatic smell. There were many processing methods in ancient times， including cleansing（water flying， removing impurities）， grinding（wine grinding， rush grinding）， frying（stir-frying， rush frying， wine frying）， degreasing， vinegar processing， decoction. In modern times， the main processing methods are simplified to cleansing， stir-frying and vinegar processing. Nowadays， the commonly used specifications include raw， fried and vinegar-processed products. Among the three specifications， raw products is the Olibanum after cleansing， fried products is a kind of Olibanum processed by frying method， vinegar-processed products is the processed products of pure frankincense mixed with vinegar. Based on the research results， it is recommended to select the resin exuded from the bark of B. carterii for the famous classical formulas such as Juanbitang containing Olibanum， processing method should be carried out in accordance with the processing requirements of the formulas， otherwise used the raw products if the formulas without clear processing requirements.

BACKGROUND: The occurrence and development of lung cancer are closely linked to epigenetic modification. Abnormal DNA methylation in the CpG island region of genes has been found in many cancers. Protein kinase C delta binding protein (PRKCDBP) is a potential tumor suppressor and its epigenetic changes are found in many human malignancies. This study investigated the possibility of PRKCDBP methylation as a potential biomarker for non-small cell lung cancer (NSCLC). METHODS: We measured the methylation levels of PRKCDBP in the three groups of NSCLC tissues. Promoter activity was measured by the dual luciferase assay, with 5'-aza-deoxycytidine to examine the effect of demethylation on the expression level of PRKCDBP. RESULTS: The methylation levels of PRKCDBP in tumor tissues and 3 cm para-tumor were higher than those of distant (>10 cm) non-tumor tissues. Receiver operating characteristic (ROC) curve analysis between tumor tissues and distant non-tumor tissues showed that the area under the line (AUC) was 0.717. Dual luciferase experiment confirmed that the promoter region was able to promote gene expression. Meanwhile, in vitro methylation of the fragment (PRKCDBP_Me) could significantly reduce the promoter activity of the fragment. Demethylation of 5'-aza-deoxycytidine in lung cancer cell lines A549 and H1299 showed a significant up-regulation of PRKCDBP mRNA levels. CONCLUSIONS PRKCDBP methylation is a potential and promising candidate biomarker for non-small cell lung cancer.
Biomarkers/metabolism* ; Carcinoma, Non-Small-Cell Lung/pathology* ; Cell Line, Tumor ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins/genetics* ; Lung Neoplasms/pathology* ; Promoter Regions, Genetic

Clinical data of 46 patients with trans-sphincter anal fistula treated by fistulectomy with external anal sphincter bareness in Department of Anorectal Surgery, Jiaxing TCM Hospital from July 2018 to July 2019 were retrospectively analyzed. All operations were performed successfully. There were no significant differences in Wexner incontinence scores (2.00±0.68 vs.1.99±0.70, P<0.05), mean anal resting pressure [(75.60±8.60) vs.(73.60±8.20)mmHg(1 mmHg=0.133 kPa), P<0.05] and maximum systolic pressure [(109.60±7.80) vs.(107.20±8.30)mmHg, P<0.05] before and 6 months after operation. There were 1 case with postoperative incision bleeding and 2 cases with postoperative infection. All patients were followed up for 6 months and there was no recurrence and changes in anal shape during the follow-up. Results indicate that the fistulectomy external anal sphincter bareness is safe, efficient with well preserved sphincter function for patients with trans-sphincter anal fistula.

Objective: To analyze the mid-term clinical efficacy of extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine. Methods: From February 2013 to December 2015, 32 patients with cervical OPLL who were extended to C1, 2 posterior open-door laminoplasty were enrolled in our hospital (complete follow-up of 25 patients). There were 19 males and 6 females, aged 48-76 years (mean 60.1±7.0 years). Cervical spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) spinal cord function score, and the rate of improvement in neurological function was calculated. The quality of life of patients was assessed using the neck disability index (NDI). Axial symptoms were assessed and indexed using the visual analog scale (VAS). The C0-2 angle of the cervical spine was measured by X-ray preoperative and at the last follow-up. The cervical curvature was evaluated by the C2-7 angle, and the cervical vertebra activity was observed by the dynamic X-ray of the cervical spine. Results: 25 patients were followed up for a period of 26-64 months (mean 35.9±8.1 months). At the last follow-up, the JOA score was 14.32±3.24, the NDI score was 7.61±1.23, and the VAS score was 1.42±0.78. The differences between the three scores were statistically significant compared with the preoperative scores. The neurological improvement rate at the last follow-up was 79.61%±13.23%. The preoperative C0-2 angle was 26.04°±6.28°, and the last follow-up was 24.92°±5.51°; the C2-7 angle was 19.55°±9.42° before surgery, and the C2-7 angle at last follow-up was 17.97°±8.80°. The C2-7 angle at last follow-up was slightly reduced compared with that preoperative, but the difference was not statistically significant. The preoperative cervical vertebra ROM was 35.31°±12.24°, and at the last follow-up it was 32.23°±9.65°. The ROM of cervical vertebrae at the last follow-up was slightly lower than that before surgery, but the difference was not statistically significant. Among them, the reduction of the ROM of overflexion was greater than that of the over-extension, which was the main reason for the decrease of ROM. The OPLL bone mass continued to grow at the last follow-up of 11 patients. Conclusion The extended to C1, 2 posterior single-door laminoplasty for the treatment of ossification of the posterior longitudinal ligament combined the upper cervical spine can achieve adequate spinal cord decompression, satisfactory neurological improvement, and improve the postoperative cervical curvature and activity. There was no obvious change in the curvature and activity of the cervical spine, and the axial symptoms did not increase significantly, and the clinical efficacy was positive.

Objective To study the pinoresinol diglucoside (PDG) on gene regulation role of ESF-1 cells in collagen secretion, to reveal PDG repair mechanisms on scalded skin.Methods The cells cultured in vitro were divided into the control group, the estradiol group and the three different PDG doses groups. The concentration of the high, medium and low dose groups were 100, 10, 1μmol/L, and that of estradiol group were 10-3μmol/L. The activity of proliferation was detected by MTT. Then collagen type I (Col I), collagen typeⅢ (ColⅢ), tissue inhibitors of metalloproteinase 1 (TIMP-1), tissue inhibitors of metalloproteinase 2 (TIMP-2) and matrix metalloproteinase 1 (MMP-1) expression levels of mRNA after administration of cells were detected by RT-PCR.Results Compared with the control group, the proliferation of ESF-1 cells (0.559 ± 0.027, 0.552 ± 0.034vs. 0.489 ± 0.027,P<0.05) in the estradiol and medium-dose PDG was significantly higher. The expression level of mRNA of ColⅠ(0.958 ± 0.021, 0.929 ± 0.031, 0.916 ± 0.015vs. 0.844 ± 0.022), ColⅢ (0.783 ± 0.038, 0.918 ± 0.021, 0.855 ± 0.017vs. 0.678 ± 0.024), TIMP-1 (0.939 ± 0.025, 0.889 ± 0.036, 0.853 ±  0.015 vs. 0.780 ± 0.023), TIMP-2 (0.507 ± 0.024, 0.655 ± 0.037, 0.572 ± 0.025vs. 0.405 ± 0.062) in the estradiol, low-, medium-dose PDG groups were significantly higher than those in the control group (P<0.05 or P<0.01). Besides, the MMP-1 (0.343 ± 0.038, 0.407 ± 0.046, 0.435 ± 0.037vs.0.519 ± 0.041) mRNA expression level in the middle and low dose PDG groups significantly decrease (P<0.05 orP<0.01). Conclusions The PDG could enhance the activity of ESF-1 cell proliferation, increase the expression of related collagen and tissue inhibitor of metalloproteinases and inhibit that of matrix metalloproteinases to repair scalded skin.

Objective To assess the treatment outcomes of anterior segment optical coherence tomography(OCT) assisted excimer laser phototherapeutic keratectomy (PTK) for fungal corneal ulcer.Methods Twenty-four patients (24 eyes) suffered from fungal corneal keratitis were collected by a retrospecive chart review with at least 6 months of follow-up data available.All corneal ulcer depth did not exceed 1/3 corneal thickness and were resistant to local and systemic medication therapy for at least 1 week.Patients were divided into 2 groups.14 patients with 14 eyes were taken PTK and 10 patients with 10 eyes were treated with lamellar keratectomy.Follow-up time was at least 6 months.Visual acuities,corneal thickness,confocal microscope,healing of the epithelial defect and inflammation and complications were recorded.Results The postoperative visual acuity in two groups were significantly improved,visual acuity of PTK treatment group improved significantly better than the control group (P ＜ 0.05).Healing time of PTK treatment group (19.43 ± 5.68) days was shorter than that of the control group (34.40 ± 5.38) days,the difference was statistically significant (P ＜0.05).There was no significant difference in the residual central corneal thickness between the PTK treatment group and the control group (P ＞ 0.05).Residual mycelium after operation in the treatment group was significantly decreased than that of control group.Healing time of epithelial in PTK treatment group (4.0 ± 1.1) days was shorter than that of the control group (6.20 ± 1.39) days,the difference was statistically significant (P ＜ 0.05).There was no corneal dilation in two groups.No recurrence in PTK treatment group was occurred.3 cases in control group had recurrence after 6 months,drug control was effective in 1 case,and other 2 cases undertook corneal transplantation.Conclusion Anterior segment OCT assisted PTK for corneal ulcer can effectively clear the lesion,accelerate epithelial healing,provide a new choice for the treatment of early superficial fungal corneal ulcer.

Objective To identify a new miRNA combination that could be used as a diagnostic marker for acute myocardial infarction (AMI) or acute cerebral infarction (ACI).Methods We surveyed the literatures and databases and selected miRNAs with a high frequency in AMI or ACI as candidate genes.We collected peripheral blood samples from 13 patients with AMI,11 patients with ACI,and 20 healthy volunteers.RNA was extracted from these samples,and gene expression levels were determined by reverse transcription and real-time quantitative PCR.The differences in the expression of the candidate miRNAs were then analyzed.Results The frequencies of miRNA1,miRNA499a,and miRNA 133a were higher in the AMI group,whereas the frequencies of let7i,miRNA16,and miRNA223 were higher in the ACI group.These six miRNAs were significantly higher in the two disease groups than in the control group (P ＜ 0.01).The levels of miRNA1 and miRNA499a were 4.82-and 3.50-times higher in the AMI group,respectively (both P ＜ 0.01),when compared with the ACI group.The levels of let7i in the ACI group were 2.61-times higher than that in the AMI group (P ＜ 0.05).There were no significant differences in the expression levels of miRNA 133a,miRNA16,and miR-NA233 between the AMI and ACI groups (P ＞ 0.05).Conclusion The abnormal increase in six miRNAs in human peripheral blood could be used as a marker for the diagnosis of AMI or ACI.The expression levels of miRNA 1 and miRNA499a were 11-to 20-times higher than control levels,and the expression levels of let7i were 6-times higher than control levels,which could be used to predict the risk of AMI and ACI,respectively.

With the advancements of stem cells and regenerative medicine, interspecies chimera has become a hot topic and will pave a new way of providing donor sources in organ transplantation. However, the interspecies chimera is confronted with a number of scientific questions and technical obstacles, including selections of appropriate embryonic stage and appropriate culture medium; those factors will deeply influence the developmental balance between donor cells and receptor embryos. Due to its relatively rapid reproductive cycle and similar organ size to human's, porcine is a very potential donor candidate to study these questions. To compare the development and chimeric efficiency of interspecies embryos, we tested and evaluated three different culture systems, PZM-3 (Porcine zygotic medium), culture medium for iPSCs (N2B27) and 3.5 h of N2B27 before PZM-3 (N2B27(3.5 h)), and two different embryonic stages, 8-cell and blastocyst in mouse-porcine chimeric embryos using parthenogenetically activated porcine embryos and mouse induced pluripotent stem cells (miPS). The results showed that, PZM-3 was beneficial for both development of chimeric embryos and miPSCs proliferation in porcine embryos in the 8-cell injection group. After early blastocyst injection, the chimeric efficiency did not appear significantly different among the three culture systems but was lower than 8-cell injection. In summary, the results suggest that 8-cell injection and PZM-3 culture medium are more beneficial to the in vitro development and chimeric efficiency of mouse-porcine chimeric embryos.
Animals ; Blastocyst ; Chimera ; Culture Media ; Embryo Culture Techniques ; veterinary ; Embryo, Mammalian ; Embryonic Development ; Induced Pluripotent Stem Cells ; cytology ; Mice ; Swine