[Saikosaponin D regulates apoptosis and autophagy of pancreatic cancer Panc-1 cells via Akt/mTOR pathway].

This study aims to investigate the effect and mechanism of saikosaponin D on the proliferation, apoptosis, and autophagy of pancreatic cancer Panc-1 cells. The cell counting kit(CCK-8) was used to examine the effects of 7, 10, 13, 16, 19, 22, 25, and 28 µmol·L~(-1) saikosaponin D on the proliferation of Panc-1 cells. Three groups including the control(0 µmol·L~(-1)), low-concentration(10 µmol·L~(-1)) saikosaponin D, and high-concentration(16 µmol·L~(-1)) saikosaponin D groups were designed. The colony formation assay was employed to measure the effect of saikosaponin D on the colony formation rate of Panc-1 cells. The cells treated with saikosaponin D were stained with hematoxylin-eosin(HE), and the changes of cell morphology were observed. Hoechst 33258 fluorescent staining was used to detect the effect of saikosaponin D on the cell apoptosis. The autophagy staining assay kit with MDC was used to examine the effect of saikosaponin D on the autophagy of Panc-1 cells. Western blot and immunocytochemistry(ICC) were employed to examine the effect of saikosaponin D on the expression levels and distribution of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cysteine-aspartic acid protease-3(caspase-3), cleaved caspase-3, autophagy-associated protein Beclin1, microtubule-associated protein light chain 3(LC3), protein kinase B(Akt), phosphorylated protein kinase B(p-Akt), mammalian target of rapamycin(mTOR), and phosphorylated mammalian target of rapamycin(p-mTOR). The results showed that compared with the control group, saikosaponin D decreased the proliferation rate of Panc-1 cells in a dose-dependent and time-dependent manner. The colony formation rate of the cells significantly decreased after saikosaponin D treatment. Compared with the control group, the cells treated with saikosaponin D became small, accompanied by the formation of apoptotic bodies. The saikosaponin D groups showed increased apoptosis rate and autophagic vesicle accumulation. Compared with the control group, saikosaponin D up-regulated the expression of Bax, cleaved caspase3, Beclin1, LC3Ⅱ/LC3Ⅰ and down-regulated the expression of Bcl-2, caspase-3, p-Akt/Akt, and p-mTOR/mTOR. In addition, these proteins mainly existed in the cytoplasm. In conclusion, saikosaponin D can inhibit the proliferation and induce the apoptosis and autophagy of Panc-1 cells via inhibiting the Akt/mTOR pathway.

[Clinical study of hydraulic perfusion pump and traditional water flushing for percutaneous endoscopic lumbar discectomy].

OBJECTIVE: To compare the clinical outcomes of hydraulic perfusion pump and traditional water fhushing in percutaneous endoscopic lumbar discectomy. METHODS: From January 2016 to December 2018, 72 patients with lumbar disc herniation failed to conservative treatment were enrolled in this study. The patients were divided into hydraulic perfusion pump group and traditional water flushing group, 36 cases in each group. There were no significant differences in gender, age, prominent segment, clinical classification, preoperative visual analogue scale (VAS) and Japanese Orthopaedic Association(JOA) score between two groups (P>0.05). All operations were performed by the same surgeon, and the operations were completed by hydraulic perfusion pumpflushing or traditional water flushing assisted surgery. The operation time, intraoperative blood loss, hospital stay were recorded;VAS and JOA scores were observed before and after operation;the incidences of postoperative complications were compared between two groups. RESULTS: All the patients were followed up for 12 to 24 (15.7±5.1) months. Compared with the traditional water flushing group, the operation time of the hydraulic perfusion pump group was shorter [(65.5±21.3) min vs (74.8±19.9) min, t=9.465, P=0.003], and intraoperative blood loss was less [(21.2±12.9)ml vs (27.4± 14.1) ml, t=8.331, P=0.012], there was no statistical difference in hospital stay between two groups. Postoperative VAS scores at 1 day, 2 weeks and the final follow-up and postoperative JOA scores at the final follow-up of the two groups of patients improved compared with those before operation, but there was no significant difference between two groups (P>0.05). In the traditional water flushing group, the intracranial hypertensionoccurred during operation and the operation time was forced to shorten in 5 cases, the nerve root adventitia injury in 4 cases, and the postoperative recurrence in 4 cases. No intracranial hypertension during operation was found in the hydraulic perfusion pump group, nerve root adventitia injury occurred in 1 case, postoperative recurrence in 2 cases. The incidence of complication of the hydraulic perfusion pump group was lower than that ofthe traditional water flushing group. CONCLUSION: Both hydraulic perfusion pump and traditional water flushing assisted percutaneous endoscopic lumbar disc herniation can achieve satisfactory clinical results, but the former has shorter operation time, clearer intraoperative vision, less bleeding, and fewer intraoperative and postoperative complications.

[Risk factors of non-surgical vertebral fracture after percutaneous kyphoplasty of single segment thoracolumbar fracture].

OBJECTIVE: To investigate the risk factors of non-surgical vertebral fracture after percutaneous kyphoplasty (PKP) in patients with single segment thoracolumbar osteoporotic vertebral compression fracture(OVCF). METHODS: The clinical data of 219 patients with single segment thoracolumbar osteoporotic vertebral compression fracture underwent percutaneous kyphoplasty from January 2010 to December 2015 were retrospectively analyzed. There were 29 cases in non-surgical vertebral body fracture group with 7 males and 22 females aged (67.35±9.92) years. Bone mineral density was(-3.89±0.72)SD, fracture compressed degree was (0.57±0.35)%, bone cement injected volume was (4.53±1.64) ml, and vertebral height recovery was (0.49±0.86)% on average. Ten cases were with vertebral body space fracture, 7 cases with bone cement leakage to the intervertebral disc, 9 cases with bone cement leakage to the peripheral tissue and 14 cases were treated by systemic anti-osteoporosis treatment after operation. Other 190 cases in non-surgical vertebral body without fracture group, and there were 42 males and 148 females, aged (66.98±7.55) years. Bone mineral density was (-3.13±0.59) SD, fracture compressed degree was (0.39±0.60)%, bone cement injected volume was (4.72±1.28) ml, and vertebral height recovery was (0.46±0.94)% on average. Among them, 13 cases were with vertebral body space fracture, 5 cases with bone cement leakage to the intervertebral disc, 29 cases with bone cement leakage to the peripheral tissue and 129 cases were treated by systemic anti-osteoporosis treatment after operation. The factors of gender, age, fracture compressed degree, fractured type, bone mineral density, bone cement injected volume, bone cement leakage, vertebral height recovery, adjacent vertebral fractures, and systemic anti-osteoporosis treatment were studied in the two groups for single factor analysis. For the results with significant differences among above factors, the multivariate logistic regression analysis was used to research the non-surgical vertebral fracture risk factors. RESULTS: Among the 219 patients with single thoracolumbar fractures, 29 cases occurred non-surgical vertebral fractures. The single factor analysis results showed that there was no significant correlation between non-surgical vertebral fracture occurrecne with age, sex, fracture compressed degree, bone cement injected volume, and vertebral height recovery(P>0.05), but there was a significant correlation between non-surgical vertebral fracture occurrence with fractured type, bone cement leakage, systemic anti-osteoporosis treatment, and bone mineral density (P<0.05). Multivariate logistic regression analysis showed that fractured type(OR=0.135), whether systemic anti-osteoporosis treatment (OR=3.793), bone cement leakage (OR=8.975), and bone density (OR=57.079) are risk factors for postoperative non-surgical vertebral fractures. CONCLUSIONS: Improving the quality of the people, systemic preventing and treating the osteoporosis, improving the surgical skills and decreasing the bone cement leakage during operation are effective methods to prevent postoperative non-surgical vertebral fractures.