Objective To establish a fast and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation.Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations` distribution,each tube of cell fraction was then partially transfered to the 96 plate well,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy.Results Three types of spermatogenic cells can be identified quickly and accurately by it`s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection.Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation is successfully developed.

Objective To discuss the effect of α-asarone on the expression level of Cyt-c,Smac,Caspase3 mRNA and protein in human esophageal carcinoma Eca-109 cell mitochondria. Methods The Eca-109 cells were cultured in vitro,and divided into the negative control group and the α-asarone treatment groups(final concentration:25,50,100 μg·mL-1).After 48 h,the morphological changes of Eca-109 cells were observed by fluorescence inversion microscope.The total RNA of cells were extracted by TRIzol method,the expressions of Cyt-c、Smac and Caspase3 were measured by RT-PCR and Western blotting. Results After Eca-109 cells were treated with different concentrations of α-asarone for 48 h,and obvious changes in the morphology were observed,the expressions of Cyt-c,Smac and Caspase3 genes and protein were increased significantly compared to the negative control group( P<0.05). Conclusion α-asarone can induce the human Eca-109 cells apoptosis by regulating expressions of mitochondrial apoptosis pathway correlation genes such as Cyt-c,Smac and Caspase3.

Objective: To analyze the clinical characteristics of streptococcal toxic shock syndrome (STSS) caused by Streptococcus pyogenes (S. pyogenes) in the third trimester to improve clinical awareness of this disease. Methods: A retrospective study was conducted to analyze the clinical data of two patients with STSS caused by S. pyogenes in Jinan Maternal and Child Health Care Hospital and nine reported cases of healthy pregnant women infected with S. pyogenes at 28-42 gestational weeks with detailed data retrieved from Chinese National Knowledge Infrastructure and PubMed database from January 1997 to December 2018. Results: The two cases admitted to our hospital were both multiparas, with onset in winter and no specific symptoms in early stage. Disseminated intravascular coagulation (DIC) was developed in both cases on admission. In one case, the fetal heart rate was reduced and disappeared soon after admission. The other patient was diagnosed as stillbirth on admission. Both patients died and blood culture revealed infection of group A streptococcus. Nine previous cases were reported in one Chinese and five English articles. No high-risk pregnancy or premature rupture of membranes was reported in these 11 cases. Among the 11 cases, ten were positive for S. pyogenes indicated by blood or tissue culture and one was positive for streptococcal toxin. Ten cases had high fever and three presented with respiratory symptoms in early stage. Abdominal pain and watery diarrhea were common symptoms and all patients developed multiple organ dysfunction and DIC. Cases occurred in winter or spring were more common. Nine women died within 36 h after the onset of fulminant symptoms. Only two survived and hospitalized for 90 d and 25 d, respectively, after emergent cesarean section indicated by reduced fetal heart rate. Unfortunately, both neonates died after birth. For the babies, there were seven intrauterine fetal deaths, one stillbirth and three live births, but only one survived (whose mother developed respiratory cardiac arrest 90 min after delivery). Conclusions STSS caused by S. pyogenes in pregnant women exacerbates rapidly with a high mortality. Early identification of clinical manifestations and rapid progress of the disease are crucial for early diagnosis and treatment, which may help improve maternal and fetal outcomes.

Under the backdrop of the "Healthy China 2030" strategy, running has become the most common form of exercise. Fitness running is a kind of endurance aerobic exercise. Compared with swimming, aerobics, and other sports with high activity and intensity, the risk of lower limb fatigue injury during fitness running is higher. In this review paper, the risk factors for running-related injuries were summarized by consulting and analyzing the database and focuses on discussing and analyzing the impact of running on joints. The results showed that hip adduction, knee bending, and ankle joint abnormalities are the three joint-related factors that cause the main injuries during running. Four rehabilitation intervention methods for running-related injuries were proposed, that can guide patients to generate personalized rehabilitation treatment plans through training.

BACKGROUND:Platelet-rich plasma has been shown to enhance the viability and the pro-angiogenesis capacity of mesenchymal stem cells.Extracellular vesicles are one of the key mediators for mesenchymal stem cells to exert their effects,but currently,it is unclear whether platelet-rich plasma affects the functions of extracellular vesicles. OBJECTIVE:To investigate the effects of platelet-rich plasma on the function of extracellular vesicles from bone marrow mesenchymal stem cells,verify whether platelet-rich plasma can be used as an adjuvant to enhance the healing effects of bone marrow mesenchymal stem cells on repairing the peripheral nerve injury. METHODS:For in vitro study,bone marrow mesenchymal stem cells were cultured under normal conditions and with 1%platelet-rich plasma.The ultracentrifugation was used to extract the extracellular vesicles produced by bone marrow mesenchymal stem cells cultured under normal conditions(EVs-nor)or the condition supplemented with 1%platelet-rich plasma(EVs-prp).Extracellular vesicles were used to incubate with Schwann cells.The EdU assay,western blot assay,qPCR and light microscopy photography were performed to examine the effects of EVs-nor and EVs-prp on Schwann cell reprogramming,which was characterized by cell proliferation,c-Jun expression,reprogramming-associated gene expression and cell morphology.For in vivo study,the model of sciatic nerve injury in rats was established.Bone marrow mesenchymal stem cells were grafted with or without 1%platelet-rich plasma into the injured rat sciatic nerve using a chitin nerve conduit.Eight weeks after the surgery,the recovery was assessed by histological and functional indexes,including regenerated nerve fiber density,gastrocnemius wet weight ratio and sciatic function index. RESULTS AND CONCLUSION:(1)Compared with EVs-nor,EVs-prp was stronger in promoting Schwann cell proliferation.The gene expressions of c-Jun and GDNF were significantly upregulated in EVs-prp treated Schwann cells.The morphology of Schwann cells was significantly longer in EVs-prp group than that in EVs-nor group,indicating that EVs-prp had a stronger ability to stimulate Schwann cell reprogramming than EVs-nor.(2)Sciatic nerve injury animal experiment results revealed that grafting mesenchymal stem cells along with platelet-rich plasma into the injured sciatic nerve showed the best recovery compared with grafting mesenchymal stem cells or platelet-rich plasma alone,demonstrated by the significantly improved density of nerve fibers,gastrocnemius wet weight ratio,and sciatic function index.(3)These results suggested that platelet-rich plasma improved the function of bone marrow mesenchymal stem cell-derived extracellular vesicles and could be served as a practical and feasible preparation to synergize with bone marrow mesenchymal stem cells to improve peripheral nerve repair.

Objective: To investigate the pathogenic characteristics of viral encephalitis in children living in Hebei province. Methods: We randomly collected cerebrospinal fluid specimens from a total of 399 children diagnosed with viral encephalitis in Hebei Children′s Hospital from May to December 2017. Real-time fluorescence quantitative PCR and Sanger sequencing were used to detect viral nucleic acids in cerebrospinal fluid by an automatic laboratory station. Statistical analysis was performed on the experimental data using SPSS 21.0 software and the clinical data were analyzed. Comparison of infection rates of EV encephalitis in different months, using line × column chi-square test. The MRI and EEG positive rates of different viral encephalitis and viral encephalitis patients not infected with the virus were analyzed by Fisher′s exact probability test. The positive rate of infection with different viruses and non-virus agents was analyzed by Fisher′s exact probability test. Results: The result showed that 80 of 399 samples were positive, and the positive rate was 20.05%. It included 22 cases of enterovirus, 4 cases of influenza A virus, 3 cases of mumps virus, 2 cases of herpes simplex virus type 1, 1 case of herpes simplex virus type 2, 4 cases of EB virus, 7 cases of cytomegalovirus, 7 cases of herpes zoster virus, 8 cases of adenovirus, 14 cases of human herpesvirus type 6. Eight cases had combined viral infection. Eight cases had concurrent infections: 3 cases had enterovirus and herpesvirus type 6 concurrent infection, 1 case had enterovirus and Japanese encephalitis virus concurrent infection and 1 case had herpes simplex virus type 2 and adenovirus, 1 case had influenza A virus herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had mumps virus and herpesvirus type 6, 1 case had herpes simplex virus type 1 and herpes zoster virus concurrent infections. Children with EV viral encephalitis in Hebei Province were highly prevalent in May and June (P=0.016). HHV6 virus encephalitis was more susceptible to infection than non-HHV6 virus (P=0.016); The rate of MRI positive findings in patients with different viral encephalitis was not statistically significant (P>0.05). The result of EEG of different viral encephalitis were P>0.05, which was not statistically significant. Conclusions EV was the most common pathogen of children with viral encephalitis in Hebei province. Encephalitis caused by influenza A virus cannot be ignored in clinical practice.