OBJECTIVETo investigate the effect of rabbit saphenous and sciatic nerve homogenates on the proliferation and calcification of rabbit osteoblasts in vitro.

METHODThe saphenous nerves (sensory nerves) and the muscular branches of the sciatic nerve (motor nerve) were collected from 48 New Zealand white rabbits to prepare the nerve tissue homogenates. Bone marrow mesenchymal stem cells (MSCs) were isolated from the rabbits and cultured in vitro, and after 14 days of routine osteogenic induction, the resultant osteoblasts were identified by immunohistochemistry, alkaline phosphatase (ALP) and Alizarin red S staining. The osteoblasts were then incubated in the induction medium containing the saphenous (sensory nerve group) or sciatic homogenates (motor nerve group), with the cells in the dexamethasone-containing, dexamethasone-free osteogenic induction medium and control medium as the control. The proliferation, total protein and ALP activity of the osteoblasts were measured every other day until the 8th day, and Alizarin red S staining was used for quantitative analysis of calcification of the cells after two weeks.

RESULTSThe application of the saphenous nerve homogenates significantly promoted cell proliferation, total protein and ALP activity (P<0.01, P<0.05 and P<0.05), while exposure of the osteoblasts to dexamethasone inhibited the cell proliferation (P<0.001). Compared to dexamethasone-free group, the saphenous homogenates enhanced the mineralization of the osteoblasts (P<0.001).

CONCLUSIONSaphenous nerve homogenates significantly promotes the proliferation, differentiation, ALP activity and mineralization of rabbit osteoblasts, but sciatic nerve homogenates do not show osteogenic effects on the cells.

Animals ; Bone Marrow Cells ; cytology ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; physiology ; Rabbits ; Sciatic Nerve ; chemistry ; Sensory Receptor Cells ; chemistry ; Tissue Extracts ; pharmacology

Phase Ⅰ clinical trials are the preliminary tests of clinical pharmacology and human safety evaluation.Majority of the subjects in phase Ⅰ clinical trials are healthy subjects.Besides,there are only a few number of healthy subjects included in one trial.Thus the data requires better accuracy and responsibility.If the subjects do not take medicine appropriately,it will directly affect the results and fail to provide accurate reference for the coming phase Ⅱ,Ⅲ clinical trial.Therefore subject medication is one of the key steps in the conduct of phase Ⅰ clinical trial.This article summarizes and discusses the key steps for oral,injection,inhalation and other delivery methods,to make sure the scientificity,accuracy and objectivity of the test results.This will also ensure the smooth development of late-stage clinical trials.

Objective To investigate the effect of glucose metabolism alteration induced by alloxan intraventricular injection on learning and memory abilities of mice, and its role in the development of AD. Methods Mice were randomly divided into high-dose alloxan intraventricular injection group (n=7, 4 mg/kg) and low-dose alloxan intraventricular injection group (n=7, 1.5 mg/kg)and control group (n=7, physiological saline); intraventricular injection of alloxan, the O-GLcNAc transferase inhibitor, was performed in the high-dose and low-dose alloxan intraventricular injection groups to interfere the brain glucose metabolism. Morris water maze was used to detect the learning and memory abilities of mice. Western blotting and immunohistochemistry were used to analyze the alterations of phosphorylation and O-Glycosylation of neurofilament in mice brain induced by alloxan intraventricular injection. Results In the located navigation tests, the swimming time and distance to find the platform in the mice of alloxan administration were significantly increased as compared with those in the control group (P＜ 0.05); in space exploration experiments, compared with those in the control mice, the number of crossing the hidden platform was decreased and the initial angle of entry to water was increased in the mice of alloxan administration (P＜0.05). Western blotting and immunohistochemistry displayed that phosphorylation was obviously increased and the O-Glycosylation was significantly reduced in the cytoskletal neurofilament of the mice with alloxan administration as compared with those in the control group (P＜0.05), which was similar to the alteration of neurofilament's modification in AD brain. Conclusion The inventricular injection of alloxan could impair the learning and memory of mice, which might have a relation with the dysregulation of phosphorylation and O-Glycosylation in neurofilament caused by the impaired glucose metabolism, which is similar to the alteration of phosphorylation and O-Glycosylation in neurofilament in AD brain.

Objective:To explore the repair methods and clinical effects of full-thickness burn wounds deep to tendon or even bone in fingers.Methods:A retrospective non-randomized controlled trial was conducted on the 98 patients with full-thickness finger burns deep to tendon or even bone who met the inclusion criteria and were hospitalized in Xiangya Hospital of Central South University from January 2010 to December 2019. Among the 98 patients, there were 81 males and 17 females, aged from 1 to 72 years, with 160 fingers involved. The wound area of each of affected fingers ranged from 2.0 cm×1.5 cm to 12.0 cm×3.5 cm, and the maximum wound area after merging the affected fingers was 12.0 cm×10.0 cm. For adult hands with multiple full-thickness burn wounds deep to tendon or even bone in multiple fingers or children with full-thickness finger burns deep to tendon or even bone, pedicled abdominal flaps were selected. For adults with single or two fingers with full-thickness burns deep to tendon or even bone, the pedicled internal hand flaps and free tissue flaps were selected. The free tissue flap repair requires good vascular conditions in the recipient area with arteries and veins available for anastomosis. For thumb nail burns deep to tendon or even bone or partial absence of the thumb after burns, the thumbs were reconstructed with the first toenail flap or dorsal foot flap with the second toe. In this study, 45 pedicled abdominal flaps were used to repair the wounds in 91 fingers, 37 pedicled internal hand flaps were used to repair the wounds in 37 fingers, 26 free tissue flaps were used to repair the wounds in 28 fingers, 3 first toenail flaps were used to reconstruct 3 patients' thumb nails and to repair hand wounds, and 1 dorsal foot flap with the second toe was used to reconstruct 1 patient's thumb and to repair hand and wrist wounds. The tissue flap area was from 2.0 cm×1.5 cm to 20.0 cm×10.0 cm. The wound in the donor site was repaired by direct suture or full-thickness skin grafting from the medial upper arm of the affected limb or split-thickness skin grafting from the outer thigh. The postoperative survival of the tissue flap, postoperative complications, and appearance and function of the flap donor site were observed. For the patients who were followed up, their finger functions were evaluated at the last follow-up using the trial criteria for replantation function evaluation of the amputated finger issued by the Hand Surgery Society of the Chinese Medical Association, and the satisfaction of the patients was investigated using the Efficacy Satisfaction Scale. Data were statistically analyzed with Kruskal-Wallis H test and Nemenyi test. Results:Of the 112 tissue flaps, 104 tissue flaps survived completely and had good blood circulation; 1 pedicled thumb dorsal ulnar reverse island flap, 1 pedicled finger artery cutaneous branch reverse island flap, and 1 free grafted anterolateral thigh perforator flap were slightly necrotic at the end, which were repaired with outer thigh split-thickness skin graft after dressing change and granulation tissue growth; 2 free grafted tarsal external artery flaps and 1 pedicled thumb dorsal ulnar reverse island flap suffered from postoperative venous return obstruction, which survived after partial suture removal and heparin saline cleansing of the wound; 1 pedicled modified dorsal metacarpal artery retrograde island flap and 1 free grafted peroneal artery perforator flap were necrotic, which were repaired by a pedicled abdominal flap and a lateral upper arm flap free transplantation respectively in stage Ⅱ. After transplantation, the tissue flaps had good shape, soft texture, and good elasticity, without bloating. There was no functional disorder in the flap donor site, and only slight scar remained. A total of 117 fingers of the 72 patients received 3-24 months of outpatient or telephone follow-up. At the last follow-up, the excellent and good rates of function evaluation of fingers repaired with pedicled abdominal flap, pedicled internal hand flap, and free tissue flap were respectively 77.3% (51/66), 96.3% (26/27), and 95.8% (23/24). The function of fingers repaired with free tissue flap and pedicled internal hand flap was significantly better than that with pedicled abdominal flap ( P<0.01). The satisfaction of patients with fingers repaired by free tissue flaps was significantly higher than that by pedicled abdominal flap ( P<0.05). Conclusions:According to the specific situation of full-thickness burn wounds deep to tendon or even bone in fingers, the pedicled abdominal flap is used to repair the multiple full-thickness burn wounds deep to tendon or even bone in multiple fingers of adult or the full-thickness burn wounds deep to tendon or even bone in fingers of children, the pedicled internal hand flap or free tissue flap is used to repair the full-thickness burn wounds deep to tendon or even bone in single or two fingers of adult patients, and the first toenail flap or the dorsal foot flap with the second toe is used to reconstruct the thumbs with full-thickness burn deep to tendon or even bone, with high postoperative tissue flap survival rate and few complications. The functional recovery of the affected finger is better after repair with free tissue flap and pedicled internal hand flap, and the patients' satisfaction is the highest after free tissue flap repair.

Objective:To study the differential expressions of piRNAs in the seminal plasma of men and the role of piRNAs in spermatogenesis.Methods:We sequenced the seminal plasma samples collected from 187 male infertility patients and 58 normal healthy men,obtained differentially expressed piRNAs,and detected the relative expressions of piRNAs in different types of sperm by RT-qPCR to explore their significance in the diagnosis of male infertility.Using histopathology,RNA-protein pull-down and Western blot,we investigated the action mechanism of piRNAs in spermatogenesis in the mouse model.Results:RT-qPCR of the seminal plasma samples revealed a high expression of hsa_piR_000478 in teratozoospermia and ROC curve analysis showed an auxiliary signifi-cance of hsa_piR_000478 in the diagnosis of the disease(AUC=0.7549).Transfection of hsa_piR_000478 and its homologous se-quence piR_mmu_54800729 into the seminiferous tubules of the mouse model significantly decreased sperm motility,increased the per-centage of morphologically abnormal sperm and destroyed the testicular structure.Molecular biological experiments exhibited a close correlation between piRNAs and the energy metabolism-related pathway,which elevated the level of cell glycolysis and interfered with normal spermatogenesis.Conclusion:hsa_piR_000478 has an auxiliary significance in the diagnosis of male infertility,and piRNAs may interfere with spermatogenesis by affecting the glycolysis-related pathway in the spermatogenic microenvironment of the testis.

Objective:To investigate the impact of ionizing radiation(IR)on the structure and function of the testis and pro-vide some strategies for the prevention and treatment of IR-induced damage(IRD).Methods:Using radiation dose simulation,se-men analysis,hormone testing,electron microscopy and single-cell transcriptome sequencing,we assessed and analyzed a case of IRD.We established a mouse model of IRD to validate the results of single-cell sequencing,and investigated the specific biological mecha-nisms of IRD and potential strategies for its intervention.Results:IR at 1-2 Gy significantly reduced sperm concentration and mo-tility,which gradually recovered after 12 months but the percentage of morphologically normal sperm remained low.It also caused im-balanced levels of various steroid hormones,decreased testosterone and dehydroepiandrosterone sulfate,increased progesterone,prolac-tin,luteinizing hormone,and follicle-stimulating hormone.Electron microscopy revealed damages to the testis structure,including loss of germ cells,atrophy of the seminiferous tubules,nuclear membrane depression of the spermatocytes,mitochondrial atrophy and de-formation,and reduction of mitochondrial cristae.Single-cell sequencing indicated significant changes in the function of the Leydig cells and macrophages and disrupted lipid-related metabolic pathways after IRD.Administration of L-carnitine to the mouse model im-proved lipid metabolism disorders and partially alleviated IRD to the germ cells.Conclusion:Ionizing radiation can cause disorders of testicular spermatogenesis and sexual hormones and inhibit lipid metabolism pathways in Leydig cells and macrophages.Improving lipid metabolism can alleviate IRD to germ cells.

BACKGROUNDAt present, a diagnostic tool with high specificity for impaired endometrial receptivity, which may lead to implantation failure, remains to be developed. We aimed to assess the different endometrial microRNA (miRNA) signatures for impaired endometrial receptivity by microarray analysis.

METHODSA total of 12 repeated implantation failure (RIF) patients and 10 infertile patients, who conceived and delivered after one embryo transfer attempt, were recruited as RIF and control groups, respectively. Endometrial specimens from the window of implantation (WOI) were collected from these two groups. MiRNA microarray was conducted on seven and five samples from the RIF and control groups, respectively. Comparative, functional, and network analyses were performed for the microarray results. Quantitative real-time polymerase chain reaction (PCR) was performed on other samples to validate the expression of specific miRNAs.

RESULTSCompared with those in the control group, the expression levels of 105 miRNAs in the RIF group were found to be significantly up- or down-regulated (at least 2-fold) by microarray analysis. The most relevant miRNA functional sets of these dysregulated miRNAs were miR-30 family, human embryonic stem cell regulation, epithelial-mesenchymal transition, and miRNA tumor suppressors by tool for annotations of microRNA analysis. Network regulatory analysis found 176 miRNA-mRNA interactions, and the top 3 core miRNAs were has-miR-4668-5p, has-miR-429, and has-miR-5088. Expression levels of the 18 selected miRNAs in new samples by real-time PCR were found to be regulated with the same trend, as the result of microarray analysis.

CONCLUSIONSThere is a significant different expression of certain miRNAs in the WOI endometrium for RIF patients. These miRNAs may contribute to impaired endometrial receptivity.

Adult ; Embryo Implantation ; genetics ; physiology ; Endometrium ; metabolism ; Female ; Humans ; Infertility, Female ; genetics ; MicroRNAs ; genetics ; Microarray Analysis ; Pregnancy ; Real-Time Polymerase Chain Reaction

Objective: To summarize the clinical characteristics of patients with Takotsubo syndrome (TTS) from China and compare these features with patients from Europe/North America. Methods: We reviewed case reports published between 1990 and 2020 with the key words of "Takotsubo syndrome" "stress cardiomyopathy" "apical balloon syndrome" and "broken heart syndrome", in Wanfang, CNKI, Pubmed and Web of Science databases, and 1 294 articles were identified, including 128 articles reporting 163 cases in China and 1 166 articles reporting 1 256 cases in Europe/North America. The characteristics of demographics, triggers, symptoms, electrocardiogram, echocardiography, left ventriculogram,coronary angiography, treatment and prognosis were analyzed and compared between Chinese and European/North American cases. Results: A total of 1 294 articles (1 419 cases: 163 from China, 1 256 from Europe/North America) were included in the final analysis. The characteristics of Chinese cases included: (1) demographic:the age was (59.6±16.9) years, which was similar with that of European/North American ((59.7±17.4) years, P=0.90), and female accounting for 78.5% (128/163), which was lower than that of European/North American (85.4% (1 073/1 256), P=0.02). (2) Triggers:mental triggers accounted for 48.5% (79/163), physical triggers accounted for 43.6% (71/163), and no triggers accounted for 7.9% (13/163), respectively. Compared with Europe/North America, the ratio of patients with mental triggers was higher in China, while the ratio of patients with physical triggers and no triggers was lower (P<0.05). (3) Symptoms: chest pain (52.8% (86/163)), chest tightness (35.0% (57/163)), shortness of breath (33.1% (54/163)), dizziness (16.0% (26/163)), sweating (15.3% (25/163)), palpitations (12.3% (20/163)), syncope (9.2% (15/163)) abdominal pain/diarrhea (8.6% (14/163)), hypotension (7.4% (12/163)), and fatigue (1.2% (2/163)) were illustrated in sequence. Compared with patients in Europe/North America, the ratio of patients with chest tightness, dizziness, sweating, palpitations, abdominal pain/diarrhea was higher in Chinese patients, while the ratio of patients with hypotension was lower in Chinese patients (P<0.05). (4) Electrocardiogram: main manifestations were myocardial ischemia symptoms, such as ST-segment elevation (63.8% (104/163)), T wave inversion (46.0% (75/163)), ST-segment depression (8.6% (14/163)). Compared with European/North American, the ratio of patients with ST-segment elevation, T wave inversion, and atrioventricular block was higher in Chinese patients (P<0.05). (5) Echocardiography and imaging:apical dyskinesia (59.5% (97/163)) and apical/left ventricular bulbar dilation (36.2%(59/163)) dominated the echocardiography findings. Compared with European/North American, the ratio of patients with apical dyskinesia, apical/left ventricular bulbar dilation, and mitral regurgitation was higher in Chinese patients, while the ratio of patients with dyskinesia in other parts and left ventricular ejection fraction<50% was lower in Chinese patients (P<0.05). Left ventricular angiography showed 36.2% (59/163) of apical dyskinesia in Chinese patients, which was higher than that reported in European/North American patients, and 38.7% (63/163) of apical/left ventricular bulbar dilation was reported in Chinese patients, which was similar to that reported in European/North American patients. Coronary angiography showed percent of no stenosis or stenosis less than 50% was 87.1% (142/163), which was similar to that reported in European/North American patients (P>0.05). The typical type of TTS accounted for 96.3% (157/163), which was significantly higher than that reported in European/ American patients, while the ratio of basal type and midventricular type was lower (P<0.01). (6) Treatment and prognosis:the applied drugs in China were listed in order as following, β-blockers (41.1% (67/163)), antiplatelet agents (37.4%(61/163)), ACEI/ARB (36.2%(59/163)), anticoagulants (27.0%(44/163)), diuretics (19.6% (32/163)), etc. Compared with Europe/North America, the ratio of antiplatelet agents, anticoagulants, statins, diuretics, and nitrates use was higher in China (P<0.05), while the use of oxygen therapy and IABP was similar (P>0.05). The hospital mortality in China was 5.5% (9/163), during 1-year follow-up the recurrence rate was 3.7% (6/163) and the mortality was 0. The prognosis was similar with that in Europe/North America. Conclusions: Compared with TTS cases in Europe/North America, TTS cases in China also occur usually in middle-aged and elderly women, most of whom have mental/physical triggers and typical imaging manifestations, followed by a low hospital mortality rate and recurrence rate.
Abdominal Pain/complications* ; Adult ; Aged ; Angiotensin Receptor Antagonists ; Angiotensin-Converting Enzyme Inhibitors ; Anticoagulants ; Arrhythmias, Cardiac/complications* ; China/epidemiology* ; Diuretics ; Dizziness/complications* ; Dyskinesias/complications* ; Electrocardiography ; Europe/epidemiology* ; Female ; Humans ; Hypotension/complications* ; Middle Aged ; Platelet Aggregation Inhibitors ; Stroke Volume ; Takotsubo Cardiomyopathy/etiology* ; Ventricular Function, Left