Objective To observe the effect of core stability training on the associated reaction of upper limbs in stroke patients with hemiplegia. Methods From November, 2014 to May, 2016, sixty stroke patients with hemiplegia were randomly divided into control group (n=30) and treatment group (n=30). Both groups accepted routine rehabilitation, while the treatment group received core stability training for 20 minutes during exercise. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), Fugl-Meyer Assess-ment-Lower Extremities (FMA-LE), Berg Balance Scale (BBS) and Associated Reaction Rating Scale (ARRS) before and 6 weeks after treatment. Results The scores of FMA-UE, FMA-LE, BBS and ARRS improved significantly in both groups after treatment (t>12.158, P<0.001), and improved more in the treatment group than in the control group (t>2.317, P<0.05). Conclusion Core stability training can re-lieve the associated reaction of upper limb in stroke patients with hemiplegia.

Objective To observe the effect of extracorporeal shock waves (ESW) therapy on avascular necrosis of femoral head (ANFH) of stage IV. Methods 72 hips of 44 patients with ANFH of stage IV, according to Association Research Circulation Osseous, were randomly divided into treatment group (n=37) and control group (n=35). The control group received routine therapy, while the treatment group received ESW in addition, 2.0-4.0 bar, 10 Hz, 6000 times. They were evaluated with Harris Hip Score, Manual Muscle Test (MMT) and Short Form of health survey (SF-36) before and 1 month after treatment. Results The scores of Harris Hip Score and MMT improved more in the treatment group than in the control group (P<0.05) after treatment, as well as the scores of SF-36 except physical health and vitality (P<0.05). Conclusion The ESW can obviously improve the motor function and quality of life of patients with ANFH of stage IV.

OBJECTIVETo observe the protective effect of combined administration of blood-activating drug and sedative drug on myocardial injury of acute myocardial infarction (AMI) rats.

METHODThe acute myocardial infarction (AMI) model was established by occluding the left descending coronary artery of Wistar rats. These rats were further divided into four groups (n = 15 per group): the sham-operated group, the AMI model group, the blood-activating drug group and the combined administration group.

RESULTCompared with the sham-operated group, the AMI model group showed significant decrease in ejection fraction (EF) and fractional shortening (FS) (P < 0.001) and notable increase in the left ventricular end-diastolic internal diameter (LVIDd) and left ventricular end-systolic internal diameter (LVIDs) (P < 0.01), with the infarct area of left ventricular front wall up to about 70%-90%. Besides, tissue was severely replaced by collagen deposition and fibrosis, the sarcomeres disorganized and mitochondrial abnormalized. Compared with the AMI model group, the blood-activating drug group and the combined administration group showed significant increase in the values of EF and FS (P < 0.05 or P < 0.01) and obvious reduction in LVIDd and LVIDs (P < 0.05 or P < 0.01), with the infarction area of left ventricular front wall up to about 40%-60%. The collagen deposition and myocardium fibrosis, the disorganized sarcomeres and mitochondrial abnormalities relieved significantly. And compared with blood-activating drug group, the combined administration group demonstrated further increase in the values of EF and FS and further decrease in LVIDd and LVIDs (P < 0.05), the collagen deposition and myocardium fibrosis, the disorganized sarcomeres and mitochondrial abnormalities relieved even more in Huoxue plus Anshen prescription group.

CONCLUSIONThe combined administration of blood-activating drug and sedative drug can further improve cardiac structure and function after myocardial ischemia infarction and have an obvious synergistic effect which may be related to sedative drug's effect of resisting lipid peroxide, stabling myocardial cell membrane and mitochondrial membrane and relieving cardiac cell injury.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Hypnotics and Sedatives ; therapeutic use ; Lipid Peroxidation ; drug effects ; Male ; Myocardial Infarction ; drug therapy ; pathology ; Rats ; Rats, Wistar

Objective: To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from different donors as first-line treatment for children and adolescents with severe aplastic anemia (SAA) . Methods: The clinical data of 79 children and adolescents with SAA diagnosed from January 2013 to December 2016 in Henan Province were retrospectively analyzed. There were 50 males and 29 females, with a median age of 14(4-18) years. 40 cases received matched sibling transplantation (MSD-HSCT), 17 with unrelated donor transplantation (UD-HSCT), and 22 with haploidentical transplantation (haplo-HSCT). Results: The comparison of MSD-HSCT, UD-HSCT, haplo-HSCT groups was conducted and the median times of neutrophils engraftment were statistically significant [12(9-25) d, 14(10-22) d, 16(11-26) d, respectively (χ²=13.302, P=0.001)], but no difference in+30 d engraftment rate [97.3%(36/37), 100%(15/15), 100%(20/20), χ²=0.959, P=0.619]. The median times of PLT engraftment were not statistically significant [14(6-34)d, 16(7-32)d, 19(10-34)d, respectively, χ²=5.892, P=0.053], and the +30 d engraftment rate had no difference [97.3%(36/37), 100%(15/15), 100%(20/20), χ²=0.959, P=0.619]. The post-transplant infection rate showed no statistically significance [35.0% (14/40), 29.4% (5/17), 45.5% (10/22), χ²=1.158, P=0.560], as well as the incidences of aGVHD, grade III/IV aGVHD and cGVHD(χ²=0.230, P=0.891; χ²=2.628, P=0.269; χ²=3.187, P=0.203). The two-years OS rate was not statistically significant respectively [(77.1±6.7)%, (70.6±11.1)%, (77.3±8.9)%, χ²=0.330, P=0.845]. Severe post-transplant infection (RR=4.617, P=0.009), grade Ⅲ/Ⅳ aGVHD (RR=2.707, P=0.048) were independent risk factors for OS. Conclusion The overall efficacy of MSD-HSCT, UD-HSCT and haplo-HSCT as first-line therapy for children and adolescents with SAA/VSAA is comparable.

ObjectiveTo explore the inhibitory effect of water extract of Broussonetiae Fructus on hepatocellular carcinoma （HCC） induced by diethyl nitrosamine （DEN） in mice based on homologous phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B （PTEN/PI3K/Akt） signaling pathway. MethodThe primary HCC mouse model was constructed by intraperitoneal injection of DEN solution， and the HCC mice were randomly divided into model group， sorafenib group （0.01 g·kg^-1·d^-1）， low-dose Broussonetiae Fructus water extract group （0.9 g·kg^-1·d^-1）， medium-dose Broussonetiae Fructus water extract group （1.8 g·kg^-1·d^-1）， and high-dose Broussonetiae Fructus water extract group （3.6 g·kg^-1·d^-1）， with 10 mice in each group. Another 10 C57BL/6 mice were selected as a control group and intraperitoneally injected with an equal volume of normal saline. Mice were treated with different concentrations of Broussonetiae Fructus water extract when liver cancer-like white nodules appeared. sorafenib group was treated with sorafenib. The control group and model group were intraperitoneally injected with normal saline. The activities of alkaline phosphatase （ALP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， and γ-glutamyl transferase （γ-GT） in the serum of mice were detected by the biochemical analyzer. The expression levels of alpha-fetoprotein （AFP） and carcinoembryonic antigen （CEA） were detected by enzyme-linked immunosorbent assay （ELISA）. The degree of hepatocyte canceration and hepatocyte injury were observed by Hematoxylin-eosin （HE） and Masson staining. The proliferation of HCC cells was observed by immunohistochemical staining. The apoptosis of HCC cells in mice was observed by erminal-deoxynucleotidyl transferase mediated nick end labelling （TUNEL） staining. The expression levels of PTEN， PI3K， Akt， and p-Akt proteins related to the PTEN/PI3K/Akt signaling pathway were detected by Western blot. ResultCompared with the control group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the model group were significantly increased （P<0.01）. Carcinogenesis and inflammatory cell infiltration were obvious in liver tissue of mice， and a large number of blue collagen fiber hyperplasia was found. The number of Ki67 positive cells was significantly increased （P<0.01）， and the expression level of PTEN protein was significantly decreased， while PI3K and p-Akt protein expression was increased （P<0.01）. Compared with the model group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the medium-dose and high-dose Broussonetiae Fructus water extract groups were significantly decreased （P<0.05， P<0.01）. The degree of carcinogenesis and inflammatory cell infiltration in liver tissue were reduced， and the collagen fiber hyperplasia was significantly reduced. The number of Ki67 positive cells was significantly decreased， and the number of TUNEL positive apoptotic cells was significantly increased （P<0.05， P<0.01）. PTEN protein expression was increased， while p-Akt protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionThe water extract of Broussonetiae Fructus has a significant inhibitory effect on DEN-induced primary HCC in mice， and its mechanism may be related to the regulation of key protein expressions in the PTEN/PI3K/Akt signaling pathway.

Humans ; Medicine, Chinese Traditional ; Colitis-Associated Neoplasms ; Drugs, Chinese Herbal/adverse effects* ; Colitis/chemically induced* ; Colorectal Neoplasms/drug therapy*

ObjectiveTo investigate the mechanism of Renshen Guben oral liquids（RGOL） in treatment of mice with renal fibrosis based on metabolomics and network pharmacology. MethodC57BL/6 mice were randomly divided into control group， model group and RGOL group， 12 mice in each group. Except for the control group， mice in the other groups were induced into unilateral ureteral obstruction（UUO） model by UUO. After preparation of the model， an aqueous solution of 4.2 g·kg^-1 extract powder was administered by gavage to RGOL group for 14 d， and an equal amount of distilled water was administered by gavage to the control and model groups. After the last administration on the 14^th day， urine was collected and detected by ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QQQ-MS/MS） with 0.1% formic acid aqueous solution as mobile phase A， and acetonitrile-isopropanol（70∶30） as mobile phase B for gradient elution（0-1 min， 5%B； 1-5 min， 5%-30%B； 5-9 min， 30%-50%B； 9-11 min， 50%-78%B； 11-13.5 min， 78%-95%B； 13.5-14 min， 95%-100%B； 14-16 min， 100%B； 16-16.1 min， 100%-5%B； 16.1-18 min， 5%B）， column temperature of 40 ℃， flow rate of 0.4 mL·min^-1， electrospray ionization（ESI）， collection range of m/z 50-900. Through network pharmacology， the targets of components in RGOL and the targets of renal fibrosis were analyzed interactively， and the key components and key targets were screened by network topology analysis， and DAVID platform was used to predict the signaling pathways of RGOL for the treatment of renal fibrosis. ResultA total of 7 differential metabolites involving 8 metabolic pathways were identified in RGOL for the treatment of renal fibrosis. The network pharmacology revealed that 36 key components in RGOL were related to 7 differential metabolites， mainly ginsenosides， notoginsenosides and nucleotides. Based on the herbs-components-targets-pathways network， a total of 23 key targets related to the treatment of renal fibrosis by RGOL were highlighted， which together with the differential metabolites were involved in linoleic acid metabolism， arginine biosynthesis， tricarboxylic acid cycle（TCA）， arginine and proline metabolism and other pathways. ConclusionBased on metabolomics and network pharmacology， this study preliminarily identified 7 differential metabolites， 36 potential pharmacodynamic components and 23 key targets and 4 key pathways in RGOL for the treatment of renal fibrosis， providing an experimental basis for the clinical application and mechanism study of this preparation.

Generation of mutants with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is commonly carried out in fish species by co-injecting a mixture of Cas9 messenger RNA (mRNA) or protein and transcribed guide RNA (gRNA). However, the appropriate expression system to produce functional gRNAs in fish embryos and cells is rarely present. In this study, we employed a poly-transfer RNA (tRNA)-gRNA (PTG) system driven by cytomegalovirus (CMV) promoter to target the medaka (Oryzias latipes) endogenous gene tyrosinase(tyr) or paired box 6.1 (pax6.1) and illustrated its function in a medaka cell line and embryos. The PTG system was combined with the CRISPR/Cas9 system under high levels of promoter to successfully induce gene editing in medaka. This is a valuable step forward in potential application of the CRISPR/Cas9 system in medaka and other teleosts.
Animals ; CRISPR-Cas Systems ; Cell Line ; Gene Editing ; Oryzias/genetics* ; RNA, Guide/genetics* ; RNA, Transfer/genetics*