We prepared protoplasts from Salvia miltiorrhiza Bunge suspension culture cells. Then, the protoplasts' vitality and functions were tested by fluorescein diacetate staining method and Fluo-3/AM flourescent probe. The optimal condition of protoplast isolation was Cellulase R-10 1.5%, Pectinase Y-23 0.3%, Macerozyme R-10 0.5%, 40 r/min 12 h, 600 r/min 5 min, and the protoplasts yield was 1.1x10(6) cells/g FW, the vitality was more than 95% by using fluorescein diacetate staining method. It has been confirmed that calcium fluorescent probe Fluo-3/AM can be successfully loaded into protoplasts.
Aniline Compounds ; chemistry ; Cell Culture Techniques ; Cellulase ; chemistry ; Fluorescent Dyes ; chemistry ; Protoplasts ; chemistry ; Salvia miltiorrhiza ; growth & development ; Xanthenes ; chemistry

BACKGROUND:There is no effective treatment for muscle atrophy caused by peripheral nerve damage. Skeletal muscle cel s, a structural unit of muscle contraction, can be used for studies on muscle atrophy when cultured in vitro.
OBJECTIVE:To investigate the promotion effect of neuron-secreted factors on the growth of skeletal muscle cel s in vitro.
METHODS:Skeletal muscle cel s primary cultured in vitro were divided into two groups:experimental group with neuron-secreted factors, and control group with common culture medium, respectively. Afterwards, the number of skeletal muscle cel s and expression level of alpha actin were detected by hematoxylin-eosin staining and immunohistochemical staining, respectively.
RESULTS AND CONCLUSION:The number of skeletal muscle cel s and expression level of alpha actin in the experimental group were significantly higher than those in the control group (P<0.05). In conclusion, neuron-secreted factors have the ability of promoting the growth of skeletal muscle cel s and may be helpful for denervated muscle atrophy.

Objective To evaluate the correlations of vascular structures,hemodynamic changes and recanalization before receiving carotid endarterectomy ( CEA) in patients with subtotal or complete occlusion of carotid artery using color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) ultrasonography. Methods A total of 107 patients were diagnosed as subtotal ( stenosis rate 95% to 99%) or complete occlusion of carotid artery with DSA and treated with CEA at Beijing Xuanwu Hospital, Capital Medical University from January 2005 to January 2014 were enrolled retrospectively. The mean age of patients was 61 ± 9 years. According to the findings of DSA,they were divided into either a carotid artery subtotal occlusion group (n=63) or a complete occlusion group (n=44). The vascular diameter,the locations of the lesions ( internal carotid artery or common carotid artery) ,the lumen echo characteristics,and whether internal-external artery collateral circulation patent or not at different stages in patients of both groups were documented. Results The lumen diameter of distal segment was significant wider in patients of the complete occlusion group compared with the subtotal occlusion group (4. 1 ± 1. 1 mm vs. 3. 2 ± 0. 8 mm). There was significant difference between the 2 groups (P <0. 01). There was no significant difference between the location of occlusion and the recanalization rate (P=0. 460). The recanalization rate of the lumen homogeneous echo ( hypoecho and echodense) filling patients (94. 1% vs. 86. 7%) was significantly higher than that of the patients of heterogeneity echo. In patients with complete occlusion of internal carotid artery,the recanalization of CEA would increase when the internal-external collateral arteries were patent. For general comparison,the recanalization rate of the subtotal occlusion group was significantly higher than that of the complete occlusion group (P<0. 01). Conclusion The carotid artery diameter normal or broadening ,the homogeneous echo in the occlusive lumen and the internal-external collateral arteries patency are closely associated with the recanalization rate. The preoperative ultrasonography has great value for the assessment of recanalization of carotid artery occlusive disease after CEA.

Objective:To investigate the expression of Col9a1 gene in ankle tissue of rat model of idiopathic congenital talipes equinovarus (ICTEV) . Methods:ICTEV rat model was established. Immunofluorescence staining and immunohistochemical dyeing technology were used to detect the expression of Col9a1 protein in ankle tissue of model group and control group. Results:The expression level of Col9a1 protein in ankle tissue in model group was higher than that in the control group (P<0.05),Col9a1 gene expressed primarily in soft tissue and cartilage membrane. Conclusion:The increasing expression level of Col9a1 gene in ankle tissue of ICTEV rats may be related to clubfoot deformity.

Objective Combining the concept of double loop learning in enterprise management with individualized nursing practice,the application value of this method in nursing care of patients with early onset severe preeclampsia(ESPE)was analyzed. Methods Patients with ESPE were selected from January 2014 to December 2016, which were divided into observation group and control group by random digits table method. The patients in observation group were given individualized nursing care with the idea of double loop learning,while the patients in control group received routine nursing care. The hospital admission, maternal pregnancy outcome and nursing satisfaction of the patients were compared between two groups. Results The observation group included 96 cases and the control group included 101 cases. There was no significant difference between two groups in age, gestational age, systolic blood pressure, diastolic blood pressure, and 24 hours urine protein(P>0.05).There were 87 cases of cesarean section in the observation group,while 94 cases in the control group, and there was no significant difference(P>0.05). The gestational weeks,the incidence of neonatal asphyxia,the nursing satisfaction was(35.5±2.7)weeks,5.2%(5/96), 90.6%(87/96)in the observation group and(34.1±2.0)weeks, 15.8%(16/101),72.3%(73/101)in the control group, the difference was statistically significant(t=4.149, χ2=5.843, 10.862, P<0.05 or 0.01). Conclusions In the ESPE nursing management, the individualized nursing and double loop learning can improve the child pregnancy outcomes and nursing satisfaction.

Objective To investigate the expression of LncRNA TCONS_00003757 in papillary thyroid carcinoma ( PTC) , and to explore the effect of LncRNA TCONS_00003757 on the proliferation, migration, and invasion in TPC-1 cells. Methods Sixty cases of papillary thyroid carcinoma specimens and paired normal tissues were collected after surgery from October 2017 to March 2018 in the Tangshan Workers Hospital. Real-time PCR analysis was used to examine the expression level of LncRNA TCONS_00003757 in PTC tissues, and the association of LncRNA TCONS_00003757 expression levels with the clinicopathological data of patients was analyzed. The effects of LncRNA TCONS_00003757 and ST3GAL5 silencing on the proliferation, migration, and invasion in TPC-1 cells were assessed by MTT, scratch test, and transwell assay. Protein expression of ST3GAL5 was detected by Western blot after LncRNA TCONS_00003757 silencing. Results LncRNA TCONS_00003757 expression was increased in PTC tissues compared with that in normal tissues ( P<0. 05 ) , which was correlated with capsule invasion and lymph node metastasis( P<0.05) . LncRNA TCONS_00003757 expression in TPC-1 cells was higher than that in normal thyroid follicular epithelial Nthy-ori 3-1 cells(P<0.05). LncRNA TCONS_00003757 and ST3GAL5 silencing inhibited the proliferation, migration, and invasion of thyroid cancer TPC-1 cells, and ST3GAL5 expression was decreased after LncRNA TCONS_00003757 silencing (P<0.05). Conclusion LncRNA TCONS_00003757 expression in PTC is significantly increased and correlated with its occurrence and development. Silencing LncRNA TCONS_00003757 may inhibit the proliferation, migration, and invasion of thyroid papillary cancer cells via decreasing ST3GAL5 expression.

Background/Aims: Although the conversion from tacrolimus (TAC) to cytotoxic T-lymphocyte-associated antigen 4-immunoglobulin (CTLA4-Ig) is effective in reducing TAC-induced nephrotoxicity, it remains unclear whether CTLA4-Ig has a direct effect on TAC-induced renal injury. In this study, we evaluated the effects of CTLA4-Ig on TAC-induced renal injury in terms of oxidative stress. Methods: In vitro study was performed to assess the effect of CTLA4-Ig on TAC-induced cell death, reactive oxygen species (ROS), apoptosis, and the protein kinase B (AKT)/forkhead transcription factor (FOXO) 3 pathway in human kidney 2 cells. In the in vivo study, the effect of CTLA4-Ig on TAC-induced renal injury was evaluated using renal function, histopathology, markers of oxidative stress (8-hydroxy-2’-deoxyguanosine) and metabolites (4-hydroxy-2-hexenal, catalase, glutathione S-transferase, and glutathione reductase), and activation of the AKT/FOXO3 pathway with insulin-like growth factor 1 (IGF-1). Results: CTLA4-Ig significantly decreased cell death, ROS, and apoptosis caused by TAC. TAC treatment increased apoptotic cell death and apoptosis-related proteins (increased Bcl-2-associated X protein and caspase-3 and decreased Bcl-2), but it was reversed by CTLA4-Ig treatment. The activation of p-AKT and p-FOXO3 by TAC decreased with CTLA4-Ig treatment. TAC-induced renal dysfunction and oxidative marker levels were significantly improved by CTLA4-Ig in vivo. Concomitant IGF-1 treatment abolished the effects of CTLA4-Ig. Conclusions CTLA4-Ig has a direct protective effect on TAC-induced renal injury via the inhibition of AKT/FOXO3 pathway.

OBJECTIVETo reveal the response of content berberine in root of Coptis chinensis to different intensity of UV-B radiation, and provide the theory basis for promoting the content of berberine.

METHODFour groups of UV-B radiation were set in the experiment which included: natural light control (0 W x m(-2)), UL (0.05 W x m(-2)), UM (0.10 W x m(-2)), UH (0.20 W x m(-2)). The special photosynthesis character, PPP pathway in the primary metabolism and lyrosinase activity, the changes of berberine in the root of C. chinensis were measured under different UV-B radiation.

RESULTPhotosynthetic pigment, qN, Fo, ETR, activity of glucose-6-phosphate dehydrogenase and the content of berberine in the root of C. chinensis, all of these parameters were lower than other groups under the UH radiation. However, under the UM radiation, C. chinensis protected itself from the light UV-B radiation by promoting the power of photosynthesis and PPP pathway in order to produce more NADPH and secondary metabolites.

CONCLUSIONC. chinensis increases its photosynthetic ability and PPP pathway which can furnish more precursor of secondary metabolites and NADPH that are needed in the secondary metabolism. Furthermore, the content of berberine increases correspondingly. The research provide the example for increasing the content of berberine in C. chinensis cultivation.

Berberine ; analysis ; Coptis ; chemistry ; drug effects ; metabolism ; Glucosephosphate Dehydrogenase ; metabolism ; NADP ; metabolism ; Photosynthesis ; radiation effects ; Ultraviolet Rays

OBJECTIVEUnder various drought conditions and nitrogen application, the content of vindoline, catharanthine, vincristine and vinblastine in the leaf of Catharanthus roseus were illustrated to improve the content of alkaloid theoretically.

METHODSix groups were set in the experiment, which included: CK (natural control), CN (natural control + nitrogen), LK (low drought), LN (low drought + nitrogen), HK (high drought), HN (high drought + nitrogen) to discuss the change characteristics of total nitrogen, the activity of alkaline POD and TDC, the content of four alkaloids under the different conditions were measured.

RESULTUnder LK condition, the activity of POD, TDC were enhanced. In the early stage of stress (0-21 d), vindoline, catharanthine, vincristine and vinblastine accumulated, and reduced in the later stage (28-35 d). For all groups, adding exogenous nitrogen could improve the total content of nitrogen, vindoline and vinblastine, meanwhile the activity of POD and TDC were enhanced as well. The LN, HN treatments were beneficial to accumulating catharanthine and vinblastine.

CONCLUSIONDrought stress or additional nitrogen have an influence on both of the activities of POD and TDC, and the four alkaloids were affected as well. Thereinto, the LN condition was the most effective treatment for accumulating the four alkaloids (vindoline, catharanthine, vincristine and vinblastine), which were regulated by improve nitrogen content and enzymatic activity.

Catharanthus ; metabolism ; Nitrogen ; metabolism ; Peroxidase ; metabolism ; Stress, Physiological ; Vinblastine ; analogs & derivatives ; metabolism ; Vinca Alkaloids ; metabolism ; Vincristine ; metabolism ; Water ; metabolism

Objective To compare the difference of cutpoint and clinical significance of HbA1C for the diagnosis of abnormal glucose metabolism in two population groups with different ages.Methods According to oral glucose tolerance test(OGTT),the cutpoint and clinical significance of HbA1C for the diagnosis of type 2 diabetes and impaired glucose regulation(IGR)were investigated in the two population groups.Results The mean HbA1C of 1 064 young subjects in an academy and 1 671 aged subjects in a community were 5.31% ±0.41% and 5.79% ±0.71%,respectively.The cutpoints of HbA1C for diagnosis of diabetes were 5.7%(specificity 86.7%,sensitivity 66.7%)and 5.9%(specificity 73.8%,sensitivity 80.1%)in the two population groups,and 5.6% for diagnosis of IGR (specificity 82.8%,sensitivity 55.8%)and 5.7%(specificity 60.9%,sensitivity 64.3%),respectively.87.8%,78.7%,and 38.5% were diagnosed diabetes by current OGTT criteria at HbA1C levels of ≥5.7%,≥5.9%,and≥6.5%,IGR being 61.6%,39.6%,and 4.1%,and normal glucose tolerance being 24.4%,10.0%,and 0.4%.Conclusion The cutpoints of HbA1C for diagnosis of diabetes and IGR are different in populations with different ages and HbA1C levels.As one of diagnostic criteria for diabetes,HbA1C 6.5% with relatively higher specificity and lower sensitivity must be combined with fasting blood glucose,random blood glucose,and OGTT.