Objective To investigate the effect of plateau hypoxia on the blood-brain barrier after subarachnoid hemorrhage (SAH) in rats. Methods Adult male SD rats (n = 78) were randomly divided into 4 groups: sham group (sham), SAH model group (SAH), plateau hypoxia sham group (Hp sham) and plateau hypoxia SAH model group (Hp SAH). The rat model of plateau hypoxia was established through low-pressure simulation chamber (altitude 5000 m), and the SAH model was established by endovascular perforation method. At 24 hours after SAH, neurobehavior score and SAH grade were assessed. The morphological changes of neurons and apoptosis of nerve cells in the CA1 region of hippocampal were observed by the staining of Nissl and TUNEL. The expression of phosphorylated PI3K (p-PI3K), PI3K, phosphorylated Akt (p-Akt), Akt, phosphorylated nuclear factor κB (p-NF-κB), NF-κB, matrix metalloproteinase-9 (MMP-9), occludin and claudin-5 in hippocampal were detected by the method of Western blotting. The expression of occludin and claudin-5 proteins in the CA1 region of hippocampal were observed by immunofluorescent staining. Results At 24 hours after SAH, the neurobehavior score decreased significantly and SAH grade increased significantly in the SAH and Hp SAH group (P< 0.05). Neurobehavior score decreased significantly in the Hp SAH group compared with the SAH group (P < 0.05). In the SAH group, neurons in the CA1 region of hippocampus were atrophied and deformed, the arrangement were disordered, the number of neurons decreased significantly, and the apoptosis of nerve cells increased significantly(P< 0.05). Plateau hypoxia could aggravate the morphological damage of neurons and apoptosis of nerve cells. The expression of p-PI3K/PI3K, p-Akt/Akt, occludin and claudin-5 proteins decreased significantly, while the expression of p-NF-κB/NF-κB and MMP-9 proteins increased significantly in the SAH and Hp SAH group (P< 0.05). The expression of p-PI3K/PI3K and MMP-9 proteins increased significantly in Hp SAH group compared with the SAH group. The expression of claudin-5 protein increased significantly in Hp sham group compared with the sham group (P < 0.05). Immunofluorescent staining showed that the expression of occludin and claudin-5 proteins in the CA1 region of hippocampus decreased in the SAH group. Plateau hypoxia could further decreased the expression of occludin and claudin-5 proteins. Conclusion Plateau hypoxia aggravates blood-brain barrier disruption after subarachnoid hemorrhage in rats through inhibiting PI3K/Akt/NF-κB pathway.

R2 R3-MYB transcription factors are ubiquitous in plants, playing a role in the regulation of plant growth, development, and secondary metabolism. In this paper, the R2 R3-MYB transcription factors were identified by bioinformatics analysis of the genomic data of Erigeron breviscapus, and their gene sequences, structures, physical and chemical properties were analyzed. The functions of R2 R3-MYB transcription factors were predicted by cluster analysis. Meanwhile, the expression patterns of R2 R3-MYB transcription factors in response to hormone treatments were analyzed. A total of 108 R2 R3-MYB transcription factors, named EbMYB1-EbMYB108, were identified from the genome of E. breviscapus. Most of the R2 R3-MYB genes carried 2-4 exons. The phylogenetic tree of MYBs in E. breviscapus and Arabidopsis thaliala was constructed, which classified 234 MYBs into 30 subfamilies. The MYBs in the five MYB subfamilies of A.thaliala were clustered into independent clades, and those in E. breviscapus were clustered into four clades. The transcriptome data showed that MYB genes were differentially expressed in different tissues of E. breviscapus and in response to the treatments with exogenous hormones such as ABA, SA, and GA for different time. The transcription of 13 R2 R3-MYB genes did not change significantly, and the expression patterns of some genes were up-regulated or down-regulated with the extension of hormone treatment time. This study provides a theoretical basis for revealing the mechanisms of R2 R3-MYB transcription factors in regulating the growth and development, stress(hormone) response, and active ingredient accumulation in E. breviscapus.
Erigeron/genetics* ; Gene Expression Regulation, Plant ; Genes, myb ; Phylogeny ; Plant Proteins/metabolism* ; Transcription Factors/metabolism*

OBJECTIVE: This study aims to establish an effective and stable periodontal ligament cell line stably expressing human telomerase reverse transcriptase (hTERT) gene by using the adenovirus method. METHODS: Polymerase chain reaction (PCR) was used to amplify the full length of hTERT gene to construct recombinant adenovirus plasmid pAd-pshuttle-cmv-hTERT. Packaged adenovirus particles were used for infection of human periodontal ligament cells. The expression levels of hTERT and osteogenic genes, such as alkaline phosphatase, Runt-related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen Ⅰ mRNA, were detected by quantitative real-time PCR (qRT-PCR). The ability of osteogenic differentiation was observed by alizarin red staining, and the cell proliferation was determined by CCK-8. RESULTS: Adenovirus particles containing the hTERT gene were successfully constructed and infected with periodontal ligament cells. The infected cells were similar to normal periodontal ligament cells. The qRT-PCR results showed that hTERT and osteogenesis-associated genes were highly expressed in the periodontal ligament cell lines constructed by adenoviruses. Alizarin red staining showed that the periodontal ligament cell line had strong osteogenic differentiation capability. CCK-8 showed that the periodontal ligament cell line had strong proliferation capability. CONCLUSIONS The human periodontal ligament cell line with high efficiency and stable expression of hTERT was established by the adenovirus method, thereby providing an ideal cell line for studying the mechanism of periodontal regeneration.
Adenoviridae ; Alkaline Phosphatase ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Humans ; Osteogenesis ; Periodontal Ligament ; Telomerase

OBJECTIVE: To follow up the participants of the randomized clinical trial "Efficacy and Safety of Niaoduqing Particles () for Delaying Moderate-to-Severe Renal Dysfunction", and assess the long-term effects of Niaoduqing Particles on delaying the progression of renal dysfunction. METHODS: Participants, who had previously been randomly assigned to receive Niaoduqing Particles or placebo for 24 weeks (146 cases in each group), were invited to follow-up and all were administered Niaoduqing Particles 5 g thrice daily and 10 g before bedtime for 24 weeks. The primary endpoints were changes in baseline serum creatinine (Scr) and estimated glomerular filtration rate (eGFR) after completion of the open-label treatment period. RESULTS: After the double-blind period, the median (interquartile range) changes in Scr were 1.1 (-13.0-24.1) and 11.7 (-2.6-42.9) μmol/L for the Niaoduqing Particle and placebo groups, respectively (P=0.008), and the median changes in eGFRs were-0.2 (-4.3-2.7) and-2.21 (-5.7-0.8) mL•min•1.73 m, respectively (P=0.016). There were significant differences in the double-blind period changes in renal function between groups. After the open-label period, the median changes in Scr were 9.0 (-10.0-41.9) and 17.5 (-6.0-50.0) μmol/L for the Niaoduqing Particle and placebo groups according to baseline grouping, respectively (P=0.214), and the median changes in eGFRs were-2.3 (-6.4-1.9) and-3.7 (-7.5-1.1) mL•min•1.73 m, respectively (P=0.134). There were no statistical differences in the open-label period changes in renal function between groups. The eGFR reduction of participants who accepted Niaoduqing Particle treatment for 48 weeks was projected to 2.5 mL•min•1.73 m per year. CONCLUSION Niaoduqing Particles appear to have long-term efficacy for patients with moderate-to-severe renal dysfunction. Although there was no statistical difference, the early use of Niaoduqing Paticles seems to ameliorate the worsening of renal function. (Trial registration No. ChiCTR-TRC-12002448).
Adult ; Disease Progression ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Glomerular Filtration Rate ; drug effects ; Humans ; Kidney Diseases ; drug therapy ; physiopathology ; Male ; Middle Aged ; Outcome Assessment (Health Care)

Objective To understand the epidemic situation and the source of infection of the reemerge human rabies in Qinghai. Methods We collected the data on human rabies and the data on the cases of multi- victims bitten by the identical dog, and also the laboratory data of the nucleoprotein ( N) gene of rabies virus from the samples which were detected by reverse transcription-polymerase chain reaction (RT-PCR) and direct immunofluorescence assay (DFA) from 2012 to 2017, to describe the epidemiological characteristics of human rabies and the prevalence of rabies virus in host animals, and to explore the source of infection of reemerge human rabies. Results A total of 7 human cases were reported in 2012-2017 in Qinghai province, among which 1 was bitted by wolf, 2 were bitted by stray dogs, 3 were bitted by domestic dogs which injured by stray dogs or wolfs. A total of 892 canine brain tissue samples were collected, from which 46 positive samples were detected with the positive rate of 5.16% (95% CI：3.70%-6.61%). The positive samples were collected from the nomadic region, which were consistent had the location of the human rabies. The samples collected from the cases of multi-victims bitten by the identical dog/animal had the positive rate of 73.08%, and 4 out of 7 human rabies were exposed to the cases of multi-victims bitten by the identical dog/animal. Genetic sequencing of the rabies virus detected from canine brain tissue samples were belong to China IV lineage, which was closely related to the Arctic clade. Conclusions The reemerging rabies happened in nomadic region of Qinghai province could be a consequence of spillover from wildlife especially from wolfs. The better surveillance system covering the human, livestock and wildlife should be set up to mitigate the rabies virus spread from the wildlife.

Objective: To investigate the clinicopathological characteristics and improve the clinical treatment of prostatic small-cell carcinoma (PSCC). METHODS: We reported 2 cases of PSCC derived from prostate cancer after treated by androgen blockade and prostate electrotomy and reviewed the relevant literature. RESULTS: Two patients with PSA elevation were diagnosed with prostate cancer by prostatic puncture biopsy and treated by maximum androgen blockade, which reduced their total PSA to the normal level. Later, due to difficult urination, they both underwent prostate electrotomy, followed by chemotherapy or radiotherapy for PSCC confirmed by postoperative pathology. Nevertheless, they died at 8 to 9 months after the discovery of PSCC. CONCLUSIONS PSCC can derive from prostate cancer after treatment, which may be attributed to the pathological mutation induced by long-term endocrine therapy. PSCC is more malignant than prostate cancer, and its prognosis is poor.

Three new prenylated stilbenes, named as cajanusins A-C (1-3), and one new natural product cajanusin D (4), along with six known derivatives (5-10) were isolated from the leaves of Cajanus cajan. Their structures were fully elucidated by means of extensive spectroscopic methods and comparison with data in the reported literatures. The new compounds of 1 and 2 were evaluated for in vitro cytotoxic activities against a panel of human cancer cell lines.

A method was established for the determination of chlorpyrifos metabolites employing QuEChERS method and ultra performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry (UPLC-QTRAP). The urine samples were extracted by acetonitrile and then cleaned up with PSA and GCB. The samples were separated with the gradient elution of acetonitrile-0.2% ammonia water on ZORBAX Eclipse Plus C18column. The analytes were detected by tandem mass spectrometry under negative ion mode with electrospray ionization (ESI) source and MRM-IDA-EPI mode. Under the optimized conditions, the calibration curve was linear in range of 1.0-100.0 μg/L,and the limits of detection were 0.10-0.73 μg/L. The average recoveries were 80.3%-90.1%, and RSDs were all within 10%. The developed method was simple,sensitive,accurate,and repeatable,and could avoid false positive result of samples effectively. The established method was successfully applied to determine the exposure level of chlorpyrifos metabolites in real samples of human health risk analysis. The results showed that the maximum concentration of chlorpyrifos metabolites was 54.6 μg/L. This method provided technic support for simultaneous identification and quantification of chemicals in complex matrix.

Objective To investigate the effect of electroacupuncture on diaphragm function of chronic obstructive pulmonary disease (COPD) rats with muscular dystrophy, and to explore the regulatory mechanism. Methods Forty male rats were randomly divided into blank group, model group, electroacupuncture group, exercise group, electroacupuncture plus exercise group, 8 rats in each group. After successful establishment of COPD rat model with muscular dystrophy, the modeled rats in various intervention groups were given electroacupuncture and/or exercise treatment. After the last treatment, the changes of rat body mass were observed, the rat lung function was detected, and the mRNA expression levels of myosin heavy chains (MHC) of MHC-1, MHC-2 and diaphragmatic related signal proteins of Atrogin-1, muscle ring-finger protein-1(MuRF-1), MyoD were detected by real-time quantitative polymerase chain reaction (qPCR). Results (1) Compared with the blank group, inspiratory resistance (IR) and functional residual mass (FRC) in the model group were increased (P < 0.05) , and the dynamic lung compliance(Cydn) was decreased(P<0.05). Compared with the model group, IR and FRC in the intervention groups were decreased (P < 0.05), but the differences among the three intervention groups were insignificant(P>0.05). (2) Compared with the blank group, the mRNA expression levels of MHC-1, Atrogin-1, MuRF-1, MyoD in the model group were increased (P<0.05), and the mRNA expression level of MHC-2 was decreased (P < 0.05). Compared with the model group, the mRNA expression levels of MHC-1, Atrogin-1, MuRF-1, MyoD in the intervention groups were decreased (P < 0.05) , and the mRNA expression level of MHC-2 was increased(P<0.05). Compared with the exercise group, the mRNA expression levels of Atrogin-1, MuRF-1, MyoD in the electroacupuncture group were decreased (P<0.05), and the mRNA expression level of MHC-2 was increased (P<0.05) , but the above indexes in electroacupuncture plus exercise group showed no obvious changes(P>0.05). Conclusion Electroacupuncture can improve respiratory function of COPD rats with muscular dystrophy, and the possible mechanism is related with the increase of MHC-2 mRNA expression and with the decrease of Atrogin-1, MuRF-1, MyoD mRNA expression, which result into the regulation of ubiquitin proteasome pathway(UPP), reduction of myosin loss, and the relief of diaphragmatic atrophy.

Objective By using resting-state fMRI (r-fMRI), to observe the effect of electroacupuncture at Quchi (LI 11) and Shangjuxu (ST 37) in changing brain activity pattern in functional bowel disorders (FBD), and to provide imaging evidence for seeking the central mechanism of Quchi (LI 11) and Shangjuxu (ST 37) in treating FBD.Method Twenty patients with functional constipation were taken as the constipation group, and 20 patients with functional diarrhea were taken as the diarrhea group. The constipation group was intervened by electroacupuncture at Shangjuxu (ST 37), while the diarrhea group was intervened by electroacupuncture at Quchi (LI 11). On the next day, the two groups both received electroacupuncture at the corresponding non-meridian point. The r-fMRI data of the two groups were collected before and after the interventions. With the functional connectivity taken as the observation index and right insular lobe as the seed point, MTLAB, DPARSF and REST were adopted to obtain the brain images functionally connected with the seed point, to further analyze the difference in brain functional connectivity before and after electroacupuncture.Result After electroacupuncture at Shangjuxu (ST 37), functional constipation patients presented positive activation in left cerebellar declive, left lingual gyrus, left middle occipital gyrus, right occipital lobe, right supramarginal gyrus, right gyrus cingula and left precuneus; the right subgyrus, right insular lobe and left insular lobe were negatively evoked. After electroacupuncture at Quchi (LI 11), patients with functional diarrhea showed significant differences in left cerebellar culmen and left posterior cingulate.Conclusion Electroacupuncture at Quchi (LI 11) and Shangjuxu (ST 37) can induce changes in brain functional connectivity related to visceral hypersensitivity in FBD patients, which is possibly associated with the effect of acupuncture in improving FBD.