Objective To study the effect of lentivirus-mediated CCL5-RNAi on the biological behaviors of human breast cancer cells. Methods CCL5-specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector, pGCSIL-GFP. Human high-metastatic breast cancer cells, MDA-MB-231, were infected by CCL5-siRNA recombinant lentivirus, which was set as KD group. Cells infected with CCL5-NC was as NC group, and cells cultured was as CON group. The expression of CCL5 mRNA and protein in MDA-MB-231 cells was detected by RT-PCR and western blot, respectively. Cell growth suppression and cell cycle was observed by MTT assay and fluorescence activated cell sorting (FACS). Colony formation and migration ability were determined by colony-rorming assay and Boyden chamber method. Results After infection of CCL5-siRNA recombinant lentivirus, the expression level of CCL5 mRNA and protein in MDA-MB-231 cells as well as the colony formation and migration ability decreased significantly, but cell's proliferation was not affected obviously. Compared with MDA-MB-231 (0.88± 0.15) and MDA-MB-231/CCL5-NC (1.00±0.07) cells, the expression of CCL5 mRNA in MDA-MB-231/ CCL5-siRNA decreased to 0.18±0.03, P<0.01. Compared with MDA-MB-231/CCL5-NC (1.82±0.18) cells, the expression of CCL5 protein in MDA-MB-231/CCL5-siRNA decreased to 0.33±0.13, P <0.01. Colony-forming assay and Boyden chamber method showed that the colony formation and migration ability of MDA-MB-231/CCL5-siRNA decreased markedly (P<0.05). The clone count in KD group was (0.33± 0.10), which was a significant decrease from (0.97±0.09) (NC group) and (1.04±0.07) (CON group), P<0.05. The number of cells that migrated through the chamber membrane of KD group (38± 15) was less than that of NC group (77±11, P <0.05) and CON group (69±9, P <0.05). However, MTT assay and FACS revealed that the proliferation of MDA-MB-231/CCL5-siRNA was not different from MDA-MB-231/CCL5-NC and MDA-MB-231 (P>0.05), the proliferation index (PI) of group KD, NC and CON were (0.48±0.02), (0.44±0.05) and (0.47±0.02) respectively. The difference was not statistically significant by multiple comparison (P>0.05). Conclusion CCL5-specific siRNA can specifically suppress the colony formation and migration of human high-matastatic breast cancer cells.

Objective To explore the adverse pregnancy outcomes of gestational diabetes mellitus (GDM) patients with different conception methods,and to construct a risk prediction model.Methods A ret-rospective cohort study was carried out to analyze the clinical data of 1453 cases of GDM patients who re-ceived regular prenatal care and delivery in the obstetrics department of the First People's Hospital of Yunnan Province from January 2018 to August 2022.A total of 436 patients with GDM who underwent in vitro fertili-zation-embryo transfer (IVF-ET) were served as the observation group,1017 GDM patients with natural ges-tation during the same period were selected as the control group.Chi-square test and t-test were used to com-pare the basic data,weight change,blood glucose reaching standard rate,maternal and infant pregnancy out-comes and related complications between the two group.The risk factors causing adverse pregnant outcomes were analyzed after controlling the confounding variables by adopting multivariate logistic regression.The pre-diction model was constructed according to the regression coefficient weight of each risk factor.The receiver operating characteristic (ROC) curve and the area under the curve (AUC) were used to evaluate the predictive efficiency of each independent risk factor and combined predictive factors.Results IVF-ET was an independ-ent risk factor for the first cesarean section (OR=1.809,95%CI:1.339-2.442),preterm birth (OR=1.622,95%CI:1.090-2.413),postpartum hemorrhage (OR=2.377,95%CI:1.406-4.018),adherent placenta (OR=1.971,95%CI:1.211-3.209),low birth weight infant (OR=2.232,95%CI:1.354-3.680) in the pa-tients with GDM (P<0.05).A combined prediction model was established by parity,age,fetus number,ges-tational weight gain,IVF-ET and scar uterus.The combined prediction probability was P=eLogit(P)/(1+eLogit(P)),Logit(P)=β1X1+β2X2+…+βnXn.The combined prediction efficacy of this study model for pre-term birth,cesarean section and low birth weight infants was good,AUC was approximately 0.8,which was greater than AUC of each independent risk factor.Conclusion The combined model of parity,age,fetus num-ber,gestational weight gain,IVF-ET and scar uterus has a certain predictive value for the risk of cesarean sec-tion,preterm birth and low birth weight infant in the patients with GDM.

OBJECTIVE To investigate the effect of a new variety of Sambucus williamsii Hance Yandan on the healing in rats with fractures. METHODS SD rats were randomly allocated to sham surgery group， model group， Zhonghua dieda pill group （0.54 g/kg）， wild S. williamsii group （5.4 g/kg， using raw drug dosage）， and high-， medium-， and low-dose groups of Yandan （10.8， 5.4， 2.7 g/kg， using raw drug dosage）， with 12 rats in each group. Except for the sham surgery group， the remaining groups were prepared with a femoral fracture model. Starting from the second day after surgery， each group was intubated with the corresponding drugs and distilled water once daily for 8 consecutive weeks. At 2， 4， and 8 weeks after administration， the levels of calcium， phosphorus， alkaline phosphatase （ALP）， and osteocalcin （OCN） in rat serum were detected. Micro-CT scanning was used to evaluate the morphology and bone microstructural parameters of the fractured femur [bone mineral density （BMD）， bone volume/tissue volume （BV/TV）， trabecular number （Tb.N）， trabecular separation （Tb.Sp）， trabecular thickness （Tb.Th）]， and hematoxylin-eosin staining was adopted to observe the morphological changes at the bone fracture site. RESULTS Compared with the model group， the levels of calcium in rat serum were significantly decreased （except at 4 weeks after administration）， and the levels of phosphorus， ALP， and OCN were significantly increased （P＜0.05） at 2， 4， and 8 weeks after administration in the high- dose group of Yandan； bone callus formation， connection of fracture ends， gradual blurring or disappearance of the fracture line， and opening of the marrow cavity were observed in the bone repair process， and a large amount of granulation tissue， fibroblasts， chondrocytes， new trabeculae， and new bone plates were visible at the fracture site of bone tissue； after 8 weeks of administration， BMD， BV/TV， Tb.N， and Tb.Th were significantly increased， and Tb.Sp was significantly decreased （P＜0.05）. Most of the above indicators in the wild S. williamsii Hance group showed no significant changes. CONCLUSIONS Yandan has the effect of promoting fracture healing in rats， and its effect is superior to that of wild S. williamsii.