This study was aimed to establish the method of identifying bupleurum cultivated germplasm using simple sequence repeat (SSR) molecular markers and to initially establish dataset of characteristic SSR bands to the bred cultivars or strains. From the bupleurum SSR primer pairs which were designed in previous work, 50 primer pairs were selected. Two bred strains and 4 other bupleurum cultivated germplasms were used as test materials. Primers pairs were screened with effective PCR amplification and high polymorphism. Meanwhile, conditions for PCR amplification and electrophoresis were optimized. Then, obtained SSR bands were analyzed and a clustering tree on the basis of genetic distance was constructed. The results showed that 9 SSR primer pairs can be used for identification. The suitable assay conditions were established and characteristic SSR bands were obtained for tested materials. The tested samples can be divided into 4 categories in the genetic similarity coefficient of 0.73. TheB. scorzonerifolium cultivated inHeilongjiang andChuanhongchaiNo. 1 strains were clustered as one category. ChuanbeichaiNo. 1 strain andZhongchai No. 1 cultivar clustered as another category. Cultivated germplasms fromSichuan Fengshunand Rongxian clustered as a unique category. It was concluded that the primer pairs and assay method established in the present study can be used as reference in identification of bupleurum cultivars or cultivated germplasms.

BACKGROUND:The homing ability of mesenchymal stem cells is closely associated with the effects of celltransplantation. Clarifying the mechanism of chemotaxis and migration wil contribute to enhance the clinical application of mesenchymal stem cells. OBJECTIVE:To investigate the effect of Cdc42 in the homing of human umbilical cord mesenchymal stem cells. METHODS:First, mesenchymal stem cells were isolated from human umbilical cord, and co-cultured with tumor necrosis factorα, interleukin-1β, and transforming growth factorβ. Western blot assay was used to test the level of Cdc42. Besides, Cdc42 siRNA was synthesized by chemical method to transfect the cells, and cellmigration and adhesion were measured by Transwel and Matrigel separately. Meanwhile, the activity of signal molecule, extracellular regulated protein kinase 1/2, was evaluated by western blot. RESULTS AND CONCLUSION:The results indicated that the inflammation factors induced the highly expression of Cdc42 in human umbilical cord mesenchymal stem cells, almost double level to controls. siRNA notably inhibited the migration and adhesion of human umbilical cord mesenchymal stem cells through Cdc42 down-regulation, and the extracellular regulated protein kinase 1/2 and phosphorylation form were also decreased simultaneously. In a word, we speculate Cdc42 plays a role in the chemotaxis of human umbilical cord mesenchymal stem cells in vitro.

Objective:To investigate the individual and combined effects of calf circumference and vitamin D levels on all-cause mortality risk in community-dwelling older adults based on the Chinese Longitudinal Healthy Longevity Survey.Methods:The calf circumference was measured in the baseline survey in 2012 and 2014.Low calf circumference was defined as <34 cm for men or 33 cm for women at screening.Vitamin D deficiency was defined as plasma 25-hydroxyvitamin D[25(OH)D]levels <50 nmol/L.All participants were followed up until 2018, when death outcomes and survival time were collected.Cox proportional hazard regression models were used to analyze the effects of calf circumference and 25(OH)D levels on the risk of all-cause mortality.Results:A total of 3 052 older adults were included in the analysis, of which 1 960(64.22%)had low calf circumference and 2 245(73.56%)had vitamin D deficiency.After 10 559.9 person-years of follow-up period, 1 312 death events were recorded.After adjusting for sociodemographic characteristics, physical activities, cognitive function, and multiple chronic diseases, calf circumference and 25(OH)D levels were negatively associated with the risk of all-cause mortality(both P<0.05). In the combined analysis, compared with the normal group, the risk of death was highest in the participants with both low calf circumference and vitamin D deficiency, which was higher than those with low calf circumference or vitamin D deficiency alone, with a hazard ratios( HR)(95% CI)of 2.51(1.81-3.45), 1.71(1.22-2.42)and 1.53(1.09-2.15), respectively.There was a significant additive interaction between low calf circumference and vitamin D deficiency on mortality(RERI>0). Conclusions:Low calf circumference and vitamin D deficiency are associated with higher mortality.Older adults with combined conditions had a even higher risk of death.Attention should be paid to joint screening and comprehensive intervention for older adults with both low calf circumference and vitamin D deficiency.

ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang ［Yiqi Yangyin Jiedu prescription （YQYYJD）］ in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor （EGFR-TKI）-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD （0， 2， 3， 4， 5， 6， 7， 8 g·L^-1） and cisplatin （0， 3， 6， 9， 12， 15， 18， 21， 24， 27 mg·L^-1） on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 （CCK-8） assay after 24 hours of intervention. The half-maximal inhibitory concentration （IC₅₀） for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group （complete medium）， YQYYJD group （5 g·L^-1）， cisplatin group （12 mg·L^-1）， and combined group （YQYYJD 5 g·L^-1 + cisplatin 12 mg·L^-1）. After 24 hours of intervention， cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay， and cell migration was evaluated by scratch and Transwell assays. Glucose consumption， lactate production， and adenosine triphosphate （ATP） levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins， including hexokinase 2 （HK2）， phosphofructokinase P （PFKP）， pyruvate kinase M2 （PKM2）， lactate dehydrogenase A （LDHA）， glucose transporter 1 （GLUT1）， and monocarboxylate transporter 4 （MCT4）， were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC₅₀ of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L^-1 and 12.91 mg·L^-1， respectively. In terms of cell proliferation， compared with the blank group， the cell survival rate and the number of colonies formed in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in cell survival rate and colony formation （P<0.01）. In terms of cell migration， compared with the blank group， the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane （P<0.01）. In terms of glycolysis， compared with the blank group， glucose consumption， lactate production， and ATP levels in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in glucose consumption， lactate production， and ATP levels （P<0.05）. Compared with the blank group， the protein expression levels of HK2， PFKP， PKM2， and LDHA in the YQYYJD， cisplatin， and combined groups were significantly decreased （P<0.01）. The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups （P<0.01）. No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes， including HK2， PFKP， PKM2， and LDHA， thereby inhibiting glycolysis.

【Objective】 To analyze the causes of a case of hemolytic disease of the fetus and newborn (HDFN),and investigate the genetic background of maternal Rh deletion D--formation. 【Methods】 Blood samples of maternal and fetus were collected, and ABO blood typing, Rh blood typing, antibody screening and identification test were performed to explore the blood group serological characteristics of Rh deletion type D--, and Rh gene sequence was performed on parturient. 【Results】 The maternal blood group was identified to be O type, D--, and the anti-Hr0 antibody against Rh high-frequency antigen was suspected to be caused by multiple pregnancies which passes through the placental barrier and enable fetus to obtain anti Hr0 antibody, leading to HDFN, with genetic testing result as RH RHCE* Ce/RHCE* Ce. 【Conclusion】 In-depth research on the formation mechanism of Rh D-- in parturient should be conducted to provide clinical value for HDFN blood exchange treatment and blood transfusion in special blood group population.