OBJECTIVE: To investigate the effect of human adipose-derived mesenchymal stem cells (hASCs) exosomes on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) extracted from osteoporotic mice, and to evaluate the effect of hASCs exosomes on preventing bone loss induced by estrogen deficiency. METHODS: hASCs exosomes were extracted by ultracentrifugation. The osteoporotic mice were established by bilateral ovariectomy (OVX). BMSCs were isolated from osteo-porotic mice and cultured for further analysis. In the experimental group, these BMSCs were exposed to an osteogenic induction medium supplemented with hASCs exosomes to evaluate their potential effects on osteogenesis. In contrast, the control group was treated with the same osteogenic induction medium, but without the addition of hASCs exosomes, to serve as a baseline comparison for the study. To comprehensively assess the osteogenic differentiation of BMSCs influenced by hASCs exosomes, alkaline phosphatase (ALP) staining, ALP activity quantitative analysis and quantitative reverse transcription polymerase chain reaction (qPCR) were performed. These evaluations provided critical insights into the role of hASCs exosomes in promoting osteoblast differentiation and bone formation in osteoporotic conditions. The fluorescence labeled hASCs exosomes were injected via the tail vein to observe the biodistribution of exosomes. Two weeks after OVX, the mice were divided into three groups: The experimental group consisted of estrogen-deficient mice receiving hASCs exosome injections; the negative control group consisted of estrogen-deficient mice receiving phosphate-buffered saline (PBS) injections; and the positive control group consisted of mice that underwent Sham surgery and received PBS injections.The injections were administered once every 3 days, for a total of 8 injections. Afterward, the femurs were collected from the mice, and micro-computed tomography (micro-CT) was performed to measure bone mineral density and conduct bone morphometric analysis. RESULTS: hASCs exosomes were successfully extracted using ultracentrifugation. After the induction by hASCs exosomes, ALP staining and ALP activity in the BMSCs extracted from osteoporotic mice were significantly enhanced, the expression of osteogenesis related genes in BMSCs were significantly up-regulated. More trabecular bone and higher bone mineral density were observed in estrogen-deficient mice injected with hASCs exosomes compared with estrogen-deficient mice injected with PBS, and there was no significant decrease in bone mineral density compared with the Sham operation group. CONCLUSION hASCs exosomes promoted the osteogenic differentiation of BMSCs extracted from osteoporotic mice. hASCs exosomes prevented bone loss induced by estrogen deficiency.
Animals ; Mesenchymal Stem Cells/cytology* ; Exosomes ; Estrogens/deficiency* ; Humans ; Osteogenesis ; Cell Differentiation ; Female ; Mice ; Osteoporosis/prevention & control* ; Ovariectomy ; Adipose Tissue/cytology* ; Cells, Cultured

Background::Differentiated thyroid cancer (DTC) is commonly diagnosed in women of child-bearing age, but whether pregnancy influences the prognosis of DTC remains controversial. This study aimed to summarize existing evidence regarding the association of pregnancy with recurrence risk in patients previously treated for DTC.Methods::We searched PubMed, Embase, Web of Science, Cochrane, and Scopus based on the prespecified protocol registered at PROSPERO (CRD42022367896). After study selection, two researchers independently extracted data from the included studies. For quantitative data synthesis, we used random-effects meta-analysis models to pool the proportion of recurrence (for pregnant women only) and odds ratio (OR; comparing the risk of recurrence between the pregnancy group and the nonpregnancy group), respectively. Then we conducted subgroup analyses to explore whether risk of recurrence differed by response to therapy status or duration of follow-up time. We also assessed quality of the included studies.Results::A total of ten studies were included. The sample size ranged from 8 to 235, with participants’ age at pregnancy or delivery ranging from 28 to 35 years. The follow-up time varied from 0.1 to 36.0 years. The pooled proportion of recurrence in all pregnant patients was 0.13 (95% confidence intervals [CI]: 0.06-0.25; I2: 0.58). Among six included studies reporting response to therapy status before pregnancy, we observed a trend for increasingly higher risk of recurrence from excellent, indeterminate, and biochemically incomplete to structurally incomplete response to therapy ( Ptrend <0.05). The pooled risk of recurrence in the pregnancy group showed no evidence of a significant difference from that in the nonpregnancy group (OR: 0.75; 95% CI: 0.45-1.23; I2: 0). The difference in follow-up time (below/above five years) was not associated with either the proportion of recurrence in all pregnant patients ( P >0.05) or the OR of recurrence in studies with a comparison group ( P >0.05). Two included studies that focused on patients with distant metastasis also did not show a significant difference in disease recurrence between pregnancy and nonpregnancy groups (OR: 0.51 [95% CI: 0.14-1.87; I2: 59%]). Conclusion::In general, pregnancy appears to have a minimal association with the disease recurrence of DTC with initial treatment. Clinicians should pay more attention to progression of DTC among pregnant women with biochemical and/or structural persistence.Registration::PROSPERO, https://www.crd.york.ac.uk/PROSPERO/; No. CRD42022367896.

Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.

This study was designed to investigate the effect of hypoxia on preeclampsia(PE)by modulating the Src/Siglec-6/SHP2 signaling pathway in the cytoplasm of trophoblast cells.Mouse model of PE was established in normal control and Siglec-6 knockdown mice by L-NAME administration,with aims of studying the changes in vascular diameter of spiral arteries in vivo and examining the expression levels of Siglec-6,p-Src,p-Shp2 and p-ERK1/2 proteins in mouse uterine vascular tissues.While,the effect of Src/Siglec-6/SHP2 on the invasive proliferation of trophoblast cells was explored by culturing human chorionic trophoblast cells HTR-8/SVneo with hypoxia in vitro.In vivo experimental assays showed that the diameter of spiral arteries was reduced in the Siglec-6 knockdown group of mice,and the expression levels of Siglec-6,p-Src,p-SHP2 and p-ERK1/2 proteins were significantly reduced.In vitro hypoxic HTR-8/SVneo cell model results revealed that Siglec-6 overexpression could promote trophoblast cell invasion and proliferation by regulating p-Src,p-SHP2,p-ERK1/2,MMP2,P53 and P21.While,suppression of Src and SHP2 eliminated Siglec-6 overexpression-mediated Siglec-6,p-Src,p-SHP2 and p-ERK1/2 expression,and inhibited the ability of Siglec-6 overexpression to mediate trophoblast invasion and proliferation.Taken together,Siglec-6 plays an important role in preeclampsia,and can alleviate preeclampsia by promoting trophoblast invasion and proliferation through the Src/SHP2 signalling pathway.

Objective:To investigate the role of RNA m6A methylation in mediating cerebellar dysplasia through analyzing the phenotypes of the mouse cerebella and the expression of several key m6A regulators upon hypobaric hypoxia treatment.Methods:Five-day old C57/BL6 mice were exposed to hypobaric hypoxia for 9 days. The status of mouse cerebellar development was analyzed by comparing the body weights, brain weights and histological features. Immunostaining of cell-type-specific markers was performed to analyze the cerebellar morphology. Real-time PCR, Western blot and immunohistochemical staining were performed to detect the expression of key m6A regulators in the mouse cerebella.Results:Compared with the control, the body weights, brain weights and cerebellar volumes of hypobaric hypoxic mice were significantly reduced ( P<0.01). The expression of specific markers in different cells, including NeuN (mature neuron), Calbindin-D28K (Purkinje cell) and GFAP (astrocyte), was decreased in hypobaric hypoxic mouse cerebella ( P<0.01), accompanied with disorganized cellular structure. The expression of methyltransferase METTL3 was significantly down-regulated in the cerebella of hypobaric hypoxic mice ( P<0.05). Conclusions:Hypobaric hypoxia stimulation causes mouse cerebellar dysplasia, with structural abnormalities in mature granular neurons, Purkinje cells and astrocytes. Expression of METTL3 is decreased in hypobaric hypoxic mice cerebellum compared with that of normobaric normoxic mice, suggesting that its mediated RNA m6A methylation may play an important role in hypobaric hypoxia-induced mouse cerebellar dysplasia.

Objective:To investigate the distribution of respiratory pathogens and risk factors of death in patients with pulmonary infection in neurosurgical intensive care unit (NICU).Methods:A total of 87 patients with pulmonary infection in the NICU of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from January 2018 to December 2019 were collected, and the pathogens of their respiratory tract were analyzed to understand the types and distribution of bacteria in the lung infection. Univariate statistical analysis was used to analyze the relationship between the patient′s clinical outcome with age, diabetes, hypertension, renal insufficiency, hypoproteinemia, anemia, chronic respiratory disease, surgery, tracheotomy, and bacterial multi-resistance. Binary logistic regression analysis was used to analyze the influencing factors of death in NICU patients with pulmonary infection.Results:A total of 112 pathogenic bacteria were isolated in this research group, including 83 Gram-negative bacteria (74.11%), 22 Gram-positive bacteria (19.64%), and 7 Fungi (5.25%). Imipenem was highly sensitive to Gram-negative bacteria, vancomycin was highly sensitive to Gram-positive bacteria, and other drugs were highly resistant. 41 patients died (47.13%). Age≥60 ( OR=3.501, 95% CI: 1.152-10.638), renal insufficiency ( OR=3.872, 95% CI: 1.336-11.224), tracheotomy ( OR=0.317, 95% CI: 0.114-0.882), bacteria multi-drug resistance ( OR=3.480, 95% CI: 1.162-10.422) were independent risk factors for death in NICU patients with pulmonary infection. Conclusions:Patients with severe neurological diseases are in critical condition, and there are many patients with pulmonary infection, with poor prognosis and high mortality. Gram-negative bacteria are the most common respiratory pathogens. Carbapenems account for the highest proportion of antibiotics in clinic. Advanced age, renal insufficiency and bacterial multidrug resistance increase the mortality of patients, while early tracheotomy can reduce the mortality of patients.

OBJECTIVE：To investigate the effects of dexmedetomidine on regional cerebral oxygen saturation and cerebral function in patients undergoing intracranial aneurysm embolization. METHODS ：Totally 44 patients undergoing intracranial aneurysm embolization in the First Affiliated Hospital of Hunan University of TCM during Jun. 2017-Aug. 2019 were collected and randomly divided into group D （22 cases）and group C （22 cases）. Ten minutes before anesthesia induction ，group D was given intravenous injection of Dexmedetomidine hydrochloride injection 1 μg/kg；group C was given buffered normal saline 20 μL. Both groups were induced with Propofol emulsion injection+Midazolam injection+Fentanyl citrate injection+Cisatracurium besylate for injection. During the operation ，group D was given Dexmedetomidine hydrochloride injection 0.5 μg（/ kg·h）+Fentanyl citrate injection+Benzsulfosum atracurium for injection+Propofol emulsion injection to maintain anesthesia ；group C was continuously pumped with buffered normal saline 0.5 μg（/ kg·h）+Fentanyl citrate injection + Benzsulfosum aratracurium for injection Propofol emulsion injection to maintain anesthesia. Before anesthesia induction （T0）， immediately after anesthesia。induction （T1）， 1 min after tracheal intubation （T2）， immediately after operation finished （T3），immediately afte extubation（T4），the mean arterial pressure（MAP），heart rate 中国药房 2021年第32卷第7期 China Pharmacy 2021Vol. 32 No. 7 ·865· （HR），regional cerebral oxygen satur ation（rSO2）were observed in 2 groups. The levels of neuron specific enolase （NSE）and S100 β protein in serum were measured at T1，T3，6 h after operation （T6）. The recovery time ，intraoperative blood loss ， nitroglycerin amount and the occurrence of ADR were recorded. RESULTS ：MAP and HR of group D at T 2-T4 were significantly lower than those at T 0；MAP and HR of group C at T 2-T4 were significantly higher than those at T 0；the group D were significantly lower than the group C at the same period （P＜0.05）；there was no statistical significance in rSO 2 between 2 groups at T 0-T4（P＞ 0.05）. The levels of serum NSE and S 100β protein in 2 groups at T 3 were significantly higher than at T 1；those in 2 groups at T 6 were significantly lower than at T 3，but those of group D were significantly lower than the group C at T 3（P＜0.05）；there was no statistical significance in the levels of serum NSE or S 100β protein between 2 groups at T 1（P＞0.05）. The recovery time of anesthesia，the amount of nitroglycerin ，the incidence of tachycardia ，nausea and vomiting ，restlessness，shivering and cough in group D were significantly shorter or lower than group C （P＜0.05）；there was no statistical significance in the intraoperative blood loss between 2 groups（P＞0.05）. CONCLUSIONS ：Dexmedetomidine can maintain the hemodynamic stability of patients with intracranial aneurysm embolization during the perioperative period ，has little effect on rSO 2 and brain function ，and has good safety.

Background & Objective: Nonsense mutations in SMC1A have been reported only in females with cluster seizures, all of whom have been described as drug-resistant epilepsy. Here, we aim to explore the use of ketogenic diet treatment. Methods: The clinical data of female patients with de novo nonsense mutations in SMC1A were collected and analyzed. The clinical data was recruited and analyzed. The peripheral blood of children and their parents was collected. The next generation sequencing was used to find suspected pathogenic mutations and all the confirmed mutations were verified by Sanger sequencing. Results: Three patients with heterozygous de novo mutations in SMC1A gene were reviewed. All patients were females, presenting with seizure onset at age between 2.5 to 11 months old. One patient had mild developmental delay. One had moderate developmental delay. Another had severe developmental retardation. None of the patients had a clinical diagnosis of Cornelia de Lange syndrome. All three patients had prominent clinical features of cluster seizures. All the nonsense mutations were predicted damaging SMC1A protein by PolyPhen-2 HVAR. All the patients were treated with multiple antiepileptic drugs but their seizures remained refractory. When initiated with ketogenic diet, they became seizure free within 3 to 4 weeks. Conclusion: SMC1A nonsense mutations can cause early onset epilepsy only in females with cluster seizures. These patients are characterized by drug-resistant epilepsy, but all our three patients have good effect on ketogenic diet add-on therapy.

Objective:To investigate the possible molecular mechanism of high-risk human papillomavirus (HPV) 16 E6 protein in promoting the proliferation, invasion and migration of cervical cancer cells.Methods:C33A cells were infected with HPV E6 protein overexpression lentivirus to construct a stable C33A-E6 cell line. The expression of heat shock protein 70 (HSP70) was detected by qRT-PCR and Western blot. Degradation rates of HSP70 at mRNA and protein levels were detected by qRT-PCR and Western blot at different time points after actinomycin D (ActD) and actinomycin ketone (CHX) treatment. Exosomes were extracted by hypervelocity centrifugation and the concentrations of HSP70 in exosomes were detected by enzyme-linked immunosorbent assay (ELISA). The concentrations of TNF-α, IL-1β, IL-6 and IL-10 in the supernatants of THP-1 cell culture that treated with exosomes isolated from C33A and C33A-E6 cells after interfering HSP70 expression were detected by ELISA. C33A cells were co-cultured with exosome-treated THP-1 cells to evaluate the changes in cell proliferation with CCK-8 method. The invasion and migration of C33A cells were assessed by Transwell assay.Results:Compared with the C33A cells, the expression of HSP70 at mRNA level in C33A-E6 cells was significantly increased ( P<0.05), but no significant change was observed at the protein level ( P>0.05). No significant difference in the degradation rate of HSP70 at mRNA or protein level was detected between C33A and C33A-E6 groups after treating with ActD and CHX for different times, but the concentration of HSP70 in exosomes was significantly increased in the C33A-E6 group ( P<0.001). The exosomes secreted by C33A-E6 cells could enhance the secretion of IL-6, TNF-α and IL-10 by THP-1 cells ( P<0.001), and the THP-1 cells treated with the exosomes could promote the proliferation, migration and invasion of C33A cells ( P<0.001). However, interfering the expression of HSP70 in C33A-E6 cells could significantly reduce the secretion of IL-6 and IL-10 by exosome-treated THP-1 cells, and inhibit the THP-1 cell-induced proliferation, migration and invasion of C33A cells ( P<0.001). Conclusions:High-risk HPV 16 E6 protein upregulated the level of HSP70 in the exosomes secreted by cervical cancer cells, thereby promoting the secretion of IL-6 and IL-10 by macrophages and enhancing the macrophage-induced proliferation, migration and invasion of cervical cancer cells.

Objective: To present the evolutionary path of Chinese Medical Ethics, analyze the development vein of periodical, explore the journal' s research focuses, and provide reference for related personnel to understand the development of the current research status in the field through visualization technology. Methods:Using biblio-metric method,taking "China National Knowledge Infrastructure" as source of data collection, we used the visual-ization software CiteSpace to draw scientific knowledge maps and analyzed literatures published from 1990 to 2014 in Chinese Medical Ethics. Results:The annualvolume of journal articles fluctuated upwards,with peaks in 1992, 2000 and 2009, and of which the most was in 2009, with 393 articles. "Medical Ethics" and "Doctor-patient Relationship" were hotwords in this field. The publications of domestic and foreign scholars promoted the interna-tional exchange and the development of Chinese medical ethics, and the most productive institutions were often col-leges or universities. Conclusion:Chinese Medical Ethics has effectively promoted the development of bioethics in China, more and more scholars are involved in the relative research of medical ethics, and the old, middle and young scientists and research teams inherit, cooperate and develop with each other. The cross-regional, inter-a-gency and interdisciplinary collaboration remains very limited, whichwill become the important factorinfluencingthe development of the field of Chinese medical ethics in the future.