The protective effect and mechanism of cornuside on diabetic nephropathy model mice / 中国药房

OBJECTIVE To investigate the protective effect and potential mechanism of cornuside on diabetic nephropathy （DN） model mice. METHODS Male KK-Ay mice were fed with high-fat and high-sugar diet for two weeks to reproduce the DN model. The successfully modeled mice were randomly grouped into model group， aminoguanidine group （positive control，100 mg/kg） and cornuside group （100 mg/kg）， and male C57BL/6J mice were included as normal group， with 6 mice in each group. Administration groups were given relevant medicine intragastrically， and normal group and model group were given a constant volume of normal saline intragastrically， once a day， for 8 consecutive weeks. The levels of fasting blood glucose （FBG）， 24 h urinary protein， serum interleukin-12 （IL-12）， IL-10， blood urea nitrogen （BUN） and serum creatinine （Scr） were detected； the pathological injury， fibrotic change and glomerular microstructure of renal tissue were observed； the expressions of the receptor of advanced glycation end products （RAGE）， collagen type Ⅳ （COL-Ⅳ） and inducible nitric oxide synthase （iNOS） in renal cortex were detected in each group. RESULTS Compared with normal group， the renal cortex of mice in model group showed obvious inflammatory cell infiltration and fibrotic changes； the mesangial hyperplasia of glomerulus was serious and the basement membrane had a large number of irregular dark dense deposits； the levels of FBG and 24 h urinary protein， the serum levels of IL- 12， BUN and Scr， and the expression levels of RAGE， COL-Ⅳ and iNOS in the renal cortex were significantly increased， while the serum level of IL-10 was significantly decreased （P＜0.01）. Compared with the model group， the renal pathological injuries， fibrotic changes and glomerular microstructure of mice in administration groups were improved significantly， and the above quantitative indexes were generally improved （P＜0.05 or P＜0.01）. CONCLUSIONS Cornuside has a certain protective effect on DN model mice. It can inhibit the inflammatory response， reduce urinary protein excretion， and alleviate renal fibrosis， which may be related to the inhibition of the advanced glycation end products/RAGE signaling pathway.

Research Advance of Chinese Medicine in Treating Atherosclerosis: Focus on Lipoprotein-Associated Phospholipase A2 / 中国结合医学杂志

As a serious cardiovascular disease, atherosclerosis (AS) causes chronic inflammation and oxidative stress in the body and poses a threat to human health. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a member of the phospholipase A2 (PLA2) family, and its elevated levels have been shown to contribute to AS. Lp-PLA2 is closely related to a variety of lipoproteins, and its role in promoting inflammatory responses and oxidative stress in AS is mainly achieved by hydrolyzing oxidized phosphatidylcholine (oxPC) to produce lysophosphatidylcholine (lysoPC). Moreover, macrophage apoptosis within plaque is promoted by localized Lp-PLA2 which also promotes plaque instability. This paper reviews those researches of Chinese medicine in treating AS via reducing Lp-PLA2 levels to guide future experimental studies and clinical applications related to AS.

Research on ability of tetramethylpyrazine on promoting neurogenesis after OGD injury based on neural stem cell microenvironment / 中国药理学通报

Aim To study the ability of tetramethylpyrazine (TMP) on promoting neurogenesis in neural stem cell microenvironment after oxygen-glucose deprivation (OGD) injury in vitro. Methods Neural stem cells (NSCs), astrocytes (ACs) and cerebral microvascular endothelial cells (BMECs) were respectively extracted and separated to establish a co-culture system. The OGD modeling conditions were optimized by NSCs activity, and the concentration of TMP was optimized by Nissl staining. Then CCK-8 and Nestin

Neuroglobin Facilitates Neuronal Oxygenation through Tropic Migration under Hypoxia or Anemia in Rat: How Does the Brain Breathe? / 神经科学通报·英文版

The discovery of neuroglobin (Ngb), a brain- or neuron-specific member of the hemoglobin family, has revolutionized our understanding of brain oxygen metabolism. Currently, how Ngb plays such a role remains far from clear. Here, we report a novel mechanism by which Ngb might facilitate neuronal oxygenation upon hypoxia or anemia. We found that Ngb was present in, co-localized to, and co-migrated with mitochondria in the cell body and neurites of neurons. Hypoxia induced a sudden and prominent migration of Ngb towards the cytoplasmic membrane (CM) or cell surface in living neurons, and this was accompanied by the mitochondria. In vivo, hypotonic and anemic hypoxia induced a reversible Ngb migration toward the CM in cerebral cortical neurons in rat brains but did not alter the expression level of Ngb or its cytoplasm/mitochondria ratio. Knock-down of Ngb by RNA interference significantly diminished respiratory succinate dehydrogenase (SDH) and ATPase activity in neuronal N2a cells. Over-expression of Ngb enhanced SDH activity in N2a cells upon hypoxia. Mutation of Ngb at its oxygen-binding site (His64) significantly increased SDH activity and reduced ATPase activity in N2a cells. Taken together, Ngb was physically and functionally linked to mitochondria. In response to an insufficient oxygen supply, Ngb migrated towards the source of oxygen to facilitate neuronal oxygenation. This novel mechanism of neuronal respiration provides new insights into the understanding and treatment of neurological diseases such as stroke and Alzheimer's disease and diseases that cause hypoxia in the brain such as anemia.

Effect of intra-operative chemotherapy with 5-fluorouracil and leucovorin on the survival of patients with colorectal cancer after radical surgery: a retrospective cohort study / 中华医学杂志(英文版)

BACKGROUND@#The effect of intra-operative chemotherapy (IOC) on the long-term survival of patients with colorectal cancer (CRC) remains unclear. In this study, we evaluated the independent effect of intra-operative infusion of 5-fluorouracil in combination with calcium folinate on the survival of CRC patients following radical resection.@*METHODS@#1820 patients were recruited, and 1263 received IOC and 557 did not. Clinical and demographic data were collected, including overall survival (OS), clinicopathological features, and treatment strategies. Risk factors for IOC-related deaths were identified using multivariate Cox proportional hazards models. A regression model was developed to analyze the independent effects of IOC.@*RESULTS@#Proportional hazard regression analysis showed that IOC (hazard ratio [HR]=0.53, 95% confidence intervals [CI] [0.43, 0.65], P  < 0.001) was a protective factor for the survival of patients. The mean overall survival time in IOC group was 82.50 (95% CI [80.52, 84.49]) months, and 71.21 (95% CI [67.92, 74.50]) months in non-IOC group. The OS in IOC-treated patients were significantly higher than non-IOC-treated patients ( P  < 0.001, log-rank test). Further analysis revealed that IOC decreased the risk of death in patients with CRC in a non-adjusted model (HR=0.53, 95% CI [0.43, 0.65], P  < 0.001), model 2 (adjusted for age and gender, HR=0.52, 95% CI [0.43, 0.64], P  < 0.001), and model 3 (adjusted for all factors, 95% CI 0.71 [0.55, 0.90], P  = 0.006). The subgroup analysis showed that the HR for the effect of IOC on survival was lower in patients with stage II (HR = 0.46, 95% CI [0.31, 0.67]) or III disease (HR=0.59, 95% CI [0.45, 0.76]), regardless of pre-operative radiotherapy (HR=0.55, 95% CI [0.45, 0.68]) or pre-operative chemotherapy (HR=0.54, 95% CI [0.44, 0.66]).@*CONCLUSIONS@#IOC is an independent factor that influences the survival of CRC patients. It improved the OS of patients with stages II and III CRC after radical surgery.@*TRIAL REGISTRATION@#chictr.org.cn, ChiCTR 2100043775.

Spatiotemporal Dynamics of the Molecular Expression Pattern and Intercellular Interactions in the Glial Scar Response to Spinal Cord Injury / 神经科学通报·英文版

Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.

Psychosocial characteristics pattern correlated with HIV-related risky sexual behavior among HIV-negative men who have sex with men: a latent profile analysis

BACKGROUND@#Men who have sex with men (MSM) have become a high risk population of HIV infection due to their risky sexual behaviors. The latent pattern of psychosocial characteristics plays an important effect in HIV-related risky behaviors among HIV-negative MSM.@*METHOD@#Participants were recruited from Wuhan, Nanchang, and Changsha city from September 2017 to January 2018. Social support was assessed by the multidimensional scale of social support, Connor-Davidson Resilience scale-10 items for reliance, the assessment of Stigma towards Homosexuality for sexual minority stigma, the Likert subscale of nondisclosure for identity concealment, the ACE questionnaire-Kaiser-CDC for adverse childhood experience, the Centers for Epidemiological Studies Depression Scale for depression. Latent profile analysis (LPA) and multivariate regression were used to analyze the data.@*RESULTS@#Three psychosocial characteristic patterns were revealed by the LPA. "Social support and resilience group" (SR group), "Identity concealment group" (IC group) and "Adverse childhood experience" (ACE group) were identified, respectively. In comparison with "SR group", "IC group" have a higher likelihood of one-night male partners (AOR = 2.74, 95%CI = [1.54, 4.90]), both fixed and one-night male partners (AOR = 2.01, 95%CI = [1.34, 3.01]) and HIV-unsure male partner (AOR = 2.12, 95%CI = [1.44, 3.13]). Similarly, "ACE group" were more likely having inconsistent condom use (AOR = 2.58, 95%CI = [1.41, 4.73]), and having sex with HIV-positive male partner (AOR = 4.90, 95%CI = [1.95, 12.30]) with comparison of "SR group". In addition, we further revealed that "ACE group" had a higher ratio (90.0%) of inconsistent condom use among MSM whose male partners were HIV-positive.@*CONCLUSIONS@#Six important psychosocial factors were divided into three latent pattern classes. Compared with "SR group", "IC group" and "ACE group" were more likely to engage in HIV-related risky sexual behaviors. Further research may pay more attention to "IC group" and "ACE group" for targeted intervention.

Advances of SHP2 modulators in the cancer immunotherapy / 药学学报

Src homology phosphotyrosyl phosphatase 2 (SHP2) is a protein tyrosine phosphatase encoded by PTPN11, which catalyzes the dephosphorylation of protein tyrosine. As a convergence node, SHP2 mediates multiple signaling pathways such as rat sarcoma (RAS)-rapidly accelerated fibrosarcoma (RAF)-mitogen-activated extracellular signal-regulated kinase (MEK)-extracellular regulated protein kinases (ERK), phosphatidylinositol 3-kinase (PI3K)-serine/threonine kinase (AKT), janus kinase (JAK)-signal transducer and activator of transcription (STAT) and programmed death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1). It can not only regulate the growth and proliferation of tumor cells, but also mediate the immune escape of tumor cells by influencing the tumor microenvironment. Given its dual biological functions in tumor immune regulation, SHP2 is a promising target for cancer immunotherapy. To date, several SHP2 allosteric inhibitors have been advanced into clinical trials for tumor immunotherapy with single or combination therapeutic strategies. Additionally, SHP2 activators also showed therapeutic potential in the field of tumor immune modulation. In this paper, we reviewed the dual function of SHP2 in both tumor and immune cells. Besides, the challenges and prospects of SHP2 modulators in cancer immunotherapy were also briefly discussed, aiming to explore new horizon of SHP2 modulators for tumor immunotherapy.

Traditional Chinese Medicine Treats Ulcerative Colitis by Intervening in Mitochondrial Quality Control： A Review / 中国实验方剂学杂志

Ulcerative colitis （UC）， a chronic inflammatory bowel disease with the accumulation of colorectal mucosa and submucosa， has a risk of developing into cancer. In recent years， the incidence of UC has been on the rise worldwide. However， its pathogenesis has not been fully elucidated by modern medicine， and even the remission rate of the latest drugs is lower than 50%， which seriously affects the patients' work and quality of life. Mitochondria， as the "power station" of eukaryotic cells， are involved in a variety of physiological processes such as the production of reactive oxygen species and the production of adenosine triphosphate by oxidative phosphorylation， and their dysfunction can lead to a series of diseases. Mitochondrial quality control （MQC） is an important way to maintain the stability of mitochondrial form， quantity， and quality. Studies have shown that MQC disorders characterized by low mitochondrial biogenesis， excessive mitochondrial oxidative stress， mitochondrial autophagy defects， mitochondrial dynamics disorders， and calcium regulation abnormalities are closely related to the occurrence and development of UC. Although progress has been achieved in the treatment of UC by traditional Chinese medicine （TCM） which can regulated MQC in a multi-pathway and multi-target manner in recent years， a review on the treatment of UC by TCM via the intervention in MQC remains to be carried out. Therefore， this paper summarized the TCM treatment of UC by regulating MQC， aiming to provide new ideas for the clinical treatment of UC by TCM.

The effect of electroacupuncture stimulation at acupoint Zusanli on the model of Staphylococcus aureus osteomyelitis / 中华创伤骨科杂志

Objective:To explore whether the electroacupunture stimulation (ES) at acupoint Zusanli (ST36) can inhibit the bone loss caused by Staphylococcus aureus (SA) infection and its mechanism in a model of SA osteomyelitis.Methods:Twelve male C57 BL/6 mice aged 10 to 12 weeks were randomly divided into 2 even groups ( n=6) for SA infection + ES or SA infection only. After ES at ST36 was conducted for 4 weeks in the model of SA osteomyelitis, samples were harvested from the femora and tibiae. Micro-CT reconstruction was performed to detect trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn.Dn) to analyze changes in bone mass. Leptin receptor (LEPR) staining was performed to detect osteoblasts. Tartrate resistant acid phosphatase (TRAP) staining was used to detect the changes in osteoclasts. The changes in plasma inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). Results:Micro-CT results showed that the BV/TV, Tb.N, Tb.Th, and Conn.Dn in the cancellous bone in the target areas in the SA + ES group were all higher than those in the SA group, LEPR immunofluorescence results indicated that the number of osteogenic precursor cells in the ES group was larger than that in the SA group, and serum ELISA indicated a decrease in inflammatory factors in the blood in the SA+ES group compared with the SA group. There were significant differences in the comparisons above ( P<0.05). There was no significant difference in the number of osteoclasts on the surface of trabecular bone between the 2 groups in TRAP staining. Conclusion:ES may slow down infectious bone destruction by inhibiting the inflammatory response induced by SA infection and by inducing aggregation and differentiation of mesenchymal stem cells into trabecular bone.

Devascularized bone surface culture for identification of microorganisms for osteomyelitis / 中华创伤骨科杂志

Objective:To evaluate devascularized bone surface culture for identification of microorganisms for osteomyelitis.Methods:A prospective study was conducted to include the eligible patients who were diagnosed with osteomyelitis and treated at Division of Orthopaedics and Traumatology, Department of Orthopaedics and Traumatology, Nanfang Hospital from December 2021 to January 2023. Their infected bone tissues were collected for both bone sample culture (BSC) and general sample culture (GSC). For BSC, the devascularized bone fragments, harvested intraoperatively, were put flat on sterile culture plates with solidified agar, their surface was gently covered with cooled and molten tryptone soy agar, and then the plates with bone samples were incubated at 37 ℃ with 5% CO 2. Meanwhile, 5 suspected samples of infected bone tissue were randomly harvested by 5 independent instruments for laboratory GSC. The culture time, bacterial species, and bacterial positive rate were compared between the 2 culture methods. Results:Included were a total of 73 patients [59 males and 14 females with an age of 49.0(31.0, 58.5) years]. The culture time for BSC [1 (1, 1) d] was significantly shorter than that for GSC [3 (2, 3) d], and the total positive rate of BSC [78.1% (57/73)] was significantly higher than that of GSC [61.6% (45/73)] ( P<0.05). The bacterial species cultured by GSC were consistent with those cultured by BSC. Conclusion:In identification of microorganisms for osteomyelitis, since BSC may be quicker and lead to a higher positive rate of bacterial culture than GSC, it can be used as an optional choice besides GCS.

Efficacy of implant surface culture in identification of microorganisms for fracture device-related infection: a preliminary analysis / 中华创伤骨科杂志

Objective:To evaluate the efficacy of implant surface culture in identification of pathogens for fracture device-related infection.Methods:A prospective study was conducted to include the eligible patients who were diagnosed with infection after fracture fixation and needed surgical removal of the implants according to treatment principles at Division of Orthopaedics and Traumatology, Department of Orthopaedics and Traumatology, Nanfang Hospital from November 2020 to January 2023. With informed consent, after rinsing with aseptic normal saline twice, their implants were gently covered with a thin layer of tryptone soy agar medium. Thereafter, the implants were incubated at 37 ℃ with 5% CO 2. Changes on the surface and in the surroundings of the implants were observed every day for consecutive 2 weeks to avoid drying up by supplementing the medium when necessary. Once pathogen colonies formed, samples were collected at 3 independent sites using sterile swabs for laboratory identification. Comparisons were made between the samples from implant surface culture and the intraoperative multisite samples from conventional culture. Results:Included were a total of 75 patients [56 males and 19 females with an age of (46.2±15.4) years]. The most common infection site was the tibia (37 cases), and the most common type of implants was plate and screw (59 cases). The positive rate of implant surface culture was significantly higher than that of conventional culture (86.7% vs. 52.0%, P<0.001). 80.5% (29/36) of the negative patients detected by the conventional culture obtained positive results by the implant surface culture; three of the positive patients detected by the conventional culture obtained negative results by the implant surface culture. The culture results were positive by both culture methods in 36 patients, and consistent by both culture methods in 35 patients, yielding a consistent rate of 97.2% (35/36). The time for implant surface culture was significantly shorter than that for conventional culture [1 (1, 2) d versus 3 (3, 4) d] ( P<0.001). Of the 65 positive patients by the implant surface culture, 59 were detected with monomicrobial infection, with Staphylococcus aureus on the top (29 cases). Conclusion:As the implant surface culture, a novel method, may be superior to the conventional culture in a significantly higher positive rate and a shorter culture time, it may be used as an effective adjunct to the conventional culture in identification of pathogens for fracture device-related infection.

Guideline for the diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients (version 2023) / 中华创伤杂志

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Guideline for clinical perioperative care of orthopedic trauma patients in the new stage of novel corona virus infection (version 2023) / 中华创伤杂志

As the National Health Commission changes the management of novel corona virus infection, the situation and preventive policies for controlling the epidemic have also entered a new stage in China. Perioperative care strategies for orthopedic trauma such as designated isolation and nucleic acid test screening have also been adjusted in the new stage. Based on the perioperative work experiences in the new stage of epidemic from the frontline anti-epidemic staff of orthopedics in domestic hospitals and combined with the literature and relevant evidence-based medical data in perioperative care of orthopedic trauma patients under the current anti-epidemic policies at home and abroad, Chinese Orthopedic Association and Chinese Society of Traumatology organized relevant experts to formulate the Guideline for clinical perioperative care of orthopedic trauma patients in the new stage of novel corona virus infection ( version 2023). The guideline summarized 16 recommendations from the aspects of preoperative diagnosis and treatment, infection prevention, emergency operation and postoperative management to systematically standardize the perioperative clinical pathways, diagnosis and treatment processes of orthopedic trauma in the new stage of novel corona virus infection, so as to provide a guidance and reference for hospitals at all levels to carry out relevant work in current epidemic control policies.

Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly (version 2023) / 中华创伤杂志

Periarticular fracture of the shoulder is a common type of fractures in the elderly. Postoperative adverse events such as internal fixation failure, humeral head ischemic necrosis and upper limb dysfunction occur frequently, which seriously endangers the exercise and health of the elderly. Compared with the fracture with normal bone mass, the osteoporotic periarticular fracture of the shoulder is complicated with slow healing and poor rehabilitation, so the clinical management becomes more difficult. At present, there is no targeted guideline or consensus for this type of fracture in China. In such context, experts from Youth Osteoporosis Group of Chinese Orthopedic Association, Orthopedic Expert Committee of Geriatrics Branch of Chinese Association of Gerontology and Geriatrics, Osteoporosis Group of Youth Committee of Chinese Association of Orthopedic Surgeons and Osteoporosis Committee of Shanghai Association of Chinese Integrative Medicine developed the Chinese expert consensus on the diagnosis and treatment of osteoporotic periarticular fracture of the shoulder in the elderly ( version 2023). Nine recommendations were put forward from the aspects of diagnosis, treatment strategies and rehabilitation of osteoporotic periarticular fracture of the shoulder, hoping to promote the standardized, systematic and personalized diagnosis and treatment concept and improve functional outcomes and quality of life in elderly patients with osteoporotic periarticular fracture of the shoulder.

Difficulties and strategies of total mesorectal excision for middle and low rectal cancer after neoadjuvant therapy / 中华消化外科杂志

Patients with local advanced rectal cancer (LARC) can benefit from neoadju-vant chemoradiotherapy (nCRT) of reducing local recurrence rate and improving survival rate. However, tissue edema after nCRT may lead to unclear tissue spaces, making it challenging for lymph node dissection and nervous system protection. The difficulty in locating inferior margin of tumor after clinical complete remission or closing to clinical complete remission, as well as the increasing risk of anastomotic leakage after nCRT, pose difficulties and new challenges of total mesorectal excision for middle and low rectal cancer. Based on literatures and clinical experiences, the authors summarize the difficulties and strategies of total mesorectal excision after nCRT, in order to provide reference for colleagues.

Dicer requiring for astrocyte quiescence in developing brains / 解剖学报

Objective To explore whether microRNAs (miRNA) in astrocytes participate in regulating the phenotypic switch in the developing central nervous system (CNS). Methods The hGFAP-CreERT; Dicer fl/fl; mT/mG and Dicer fl/fl; mT/mG mice were generated and induced by tamoxifen (TMF). The)' were divided into 7 days, 10 days, 14 days Dicer conditional knock out (Dicer CKO) group and control group according to their age and the expression of hGFAP-CreERT. These 24 mice were divided into six groups and each group contained four mice. Then the change of astrocytes in the developing CNS was observed by immunofluorescenct staining. Results Both of the densities of astrocytes and microglial cells increased in the Dicer CKO mice brains as compared to littermate controls. Conclusion Dicer and miRNA of astrocytes are important for astrocytes quiescence in developing brains.

Comparative research on separation and purification methods of neural stem cells / 中国药理学通报

Aim To explore and optimize the different separation and purification methods of neural stem cells (NSCs) , and compare the cell viability and purity after separation and purification.Methods (X) Separation of NSCs: The brains of ICR mice born within 24 h were isolated ( the cerebellum and olfactory bulb removed) , chopped and digested by mechanical pipetting, trypsin digestion, and PDD enzyme digestion.After sieving and centrifugation, the brain tissues were inoculated at a concentration of 1 X 10 • L , and cultured in serum-free medium to observe and compare the activity, proliferation and number of miscellaneous cells.(2) Purification of NSCs: The brain tissues of mice born within 24 h were isolated, and NSCs were purified by differential centrifugation and sieving, and the purity was identi-fied by immunofluorescence.Results NSCs separated by PDD enzyme digestion had high survival rate and rapid proliferation rate; the Nestin positive rate of NSCs in both Sieving assay and Differential centrifugation assay groups was higher than that of primary NSCs (59.14% ± 0.16% , P <0.05) , and the former (93.54% ± 0.02%) was higher than the latter (86.12% ± 0.04% ) .Conclusions The three separation methods can obtain a large amount of NSCs.Among them, the PDD enzymatic digestion method has little damage to the NSCs, and the neuro-sphere proliferation speed is fast; the NSCs obtained by the sieving assay have higher purity.

Chuanxiong injection improves neurological function of post-stroke depression rats through cAMP-CREB-BDNF pathway / 中国药理学通报

Aim To explore the neuroproteetion of Chuanxiong injection on post-stroke depression (PSD) and its effeet on cAMP-p-CREB-BDNF pathway in hip¬pocampus.Methods Hats were randomly divided into four groups of sham, model, low-dose ( 0.1 g • kg ~1 ) of Chuanxiong injection and high-dose (0.2 g • kg 1 ) of it.The PSD model was established by middle cere-bral artery occlusion ( MCAO) combined with chronic stress stimulation.After the treatment, the behaviors of rats were observed by the tests of sugar consumption, open-field and Morris water maze.The function and status of neurons in hippocampus CA1 area were detec¬ted by Nissl staining and Neun immunofluorescence.The expressions of cAMP-p-CREB-BDNF pathway pro¬teins in hippocampus were observed by Western blot.Results Both Chuanxiong injection groups of low-dose and high-dose significantly increased consumption of sugar water, horizontal anrl vertical movement scores, decreased total route distance and route near the plat, elevated the number of Nissl body in CA1 area, re¬stored the morphology of Nissl body, and enhanced the expressions of cAMP and p-CREB in hippocampus.Additionally, the high dose group shortened swimming duration, reduced distance ratio of near plat to far from plat, and increased the expressions of Neun and BD- NF.Conclusions Chuanxiong injection has the po¬tential of improving the behavior and neurological func¬tion of PSD rats via up-regulating cAMP-CREB-BDNF pathway, protecting the neurons in hippocampal CA1 area, and ultimately alleviating the cognitive deficiency of PSD rats.

Bioinformatics analysis on structure and function and expression vector construction of SARS-CoV-2 related protein TMPRSS2 / 中国药理学通报

Aim Human TMPRSS2 is a transmembrane serine protease.In this paper, the structure and func¬tion of the protein were systematically analyzed by bioinformatics, the codon was optimized and the pro- karvotie expression vector was constructed to explore the molecular mechanism of SARS-CoV-2 infecting host cells.Methods The recombinant expression vector pET-22b-TMPRSS2 was generated by molecular clo¬ning technology.The homology, functional sites, sub¬cellular localization, three-dimensional structure and evolutionary characteristics of TMPRSS2 protein were systematically analyzed by using analytical tools such as Protparam, NetPhos3.1, Blast, Clustal X2 and MEGA7.0.Results The prokarvotic expression plas- mid was constructed correctly; TMPRSS2 belongs to medium molecular weight protein, which is composed of 492 amino acid residues.The theoretical isoelectric point is 8.12, the molecular extinction coefficient is 118 145 L • mol~1 • cm"1 , and the half-life is 30 h; TMPRSS2 has 15 potential glycosylation sites and 49 possible phosphorylation sites.It is a transmembrane hydrophilie protein without signal sequenee.In addi¬tion, the protein has 13 potential B-cell epitopes and 7 T-eell epitopes.Seeondarv structure analysis showed that random coil accounted for the highest proportion of TMPRSS2 protein ( 0.453 3) , followed by extended strand (0.252 0).Sequence comparison and evolu¬tionary analysis showed that the highest sequence con¬sistency and closest genetic relationship with human TMPRSS2 was Pan troglodytes, followed by gorilla.Conclusions Human-derived TMPRSS2 protein is ev- olutionarilv conserved and functionally important.Hie results of this study can help to reveal the structure and mechanism of action of TMPRSS2 protein, provide ide¬as for the diagnosis and treatment of COYID-19, and accelerate the research and development process of new drugs targeting TMPRSS2 protein.