Objective To investigate the effect of diosgenin on the proliferation and apoptosis of breast cancer cells and its potential molecular mechanism. Methods The breast cancer cell line MCF-7 was treated with low, medium, and high doses of diosgenin, and cell proliferation was detected through the MMT method. Flow cytometry was used to detect cell apoptosis. Nuclear-cytoplasmic-protein separation method was applied to detect the subcellular localization of death associated protein (DAXX). qRT-PCR and Western blot were used to detect the expressions of DAXX and c-Jun N-terminal kinase pathway (JNK)-related proteins. Results Diosgenin considerably inhibited the proliferation of MCF-7 cells and promoted cell apoptosis in a concentration-dependent manner. Diosgenin can promote the movement of DAXX from nucleus into the cytoplasm. Diosgenin upregulated the expression of cell surface death receptor (Fas), increased the phosphorylation levels of JNK and mitogen activated protein kinase (p38), and activated the JNK/p38 signaling pathway with concentration dependence. Conclusion Diosgenin inhibits the proliferation and promotes the apoptosis of the breast cancer cell line MCF-7, whose mechanism may be related to the regulation of DAXX subcellular localization and the activation of JNK/p38 signaling pathway.

Objective To explore the expression of death domain-associated protein(DAXX)and its subcellular localization in MCF-7 cells,discussing their effect on cellular function of MCF-7 cells.Methods MCF-7 cells were transfected with LV-DAXX-RNAi-GFP plasmid and LV-DAXX-GFP overexpression lentiviral plasmid,DAXX was knocked down and overexpressed.The expression and subcellular localization of DAXX in MCF-7 cells were detected by nuclear-cytoplasm separation methods and immu-nofluorescence,cell apoptosis and cell cycle were detected by flow cytometry,and cell proliferation was detected by MTT method.Results Both of the nuclear-cytoplasm separation methods and immunofluorescence confirmed that DAXX was mainly located in nucleus of MCF-7 cells.The transfection of LV-DAXX-GFP lentiviral plasmid mainly up-regulated the expression of DAXX in the cytoplasm of MCF-7 cells,while the transfection of LV-DAXX-RNAi-GFP lentiviral plasmid mainly down-regulated the expression of DAXX in the nucleus of MCF-7 cells.Flow cytometry confirmed that down-regulated the expression of DAXX located in nucleus could increase the apoptosis rate of MCF-7 cells,decreasing the cell proliferation,and block the cells in S phase,while up-regulated the expression of DAXX in cytoplasm had no significant effect on cell apoptosis,cell cycle and cell proliferation of MCF-7 cells.Conclusion DAXX is mainly located in the nucleus of MCF-7 cells.The expression of DAXX in the nucleus or cytoplasm may have the opposite effect on the cellular function of MCF-7 cells.

Objective:To evaluate the role of Sigma-1 receptor (Sigma-1R) in pentazoxine-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury in SH-SY5Y cells and the relationship with endoplasmic reticulum stress (ERS).Methods:The well-growing SH-SY5Y cells were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), OGD/R group (group O), OGD/R+ pentazoxin group (group OP) and OGD/R+ pintazoxin+ BD1047 group (group OPB). The cells in group C were normally cultured. In O group, OP group and OPB group, the culture medium was replaced with EBSS medium, and then the cells were cultured in an incubator of 5% CO 2-95% N 2 at 37 ℃ for 4 h, then replaced with DMEM/F12 medium containing 10% fetal bovine serum for restoration of O 2-glucose supply for 18 h, and in addition pentazoxin (final concentration 10 μmmol/L) was added during restoration in OP group, and pentazoxin (final concentration 10 μmmol/L) and Sigma-1R blocker BD1047 (final concentration 20 μmol/L) were added during restoration in OPB group. The apoptosis rate was detected by flow cytometry at the end of restoration, and the expression of Sigma-1R, C/EBP homologous protein (CHOP), phosphorylated inositol-requiring enzyme 1 (p-IRE1), spliced X-box binding protein 1 (XBP1s), and activated caspase-3 (c-cas-3) was detected by Western blot. Results:Compared with group C, the apoptosis rate was significantly increased, and the expression of CHOP and p-IRE1 was up-regulated in O group, OP group and OPB group, the expression of XBP1s and c-cas-3 was significantly up-regulated in O group and OPB group ( P<0.05), and no significant change was found in the expression of Sigma-1R, XBP1s and c-cas-3 in OP group ( P>0.05). Compared with O group, the apoptosis rate was significantly decreased, the expression of Sigma-1R was up-regulated, and the expression of CHOP, p-IRE1, XBP1s and c-cas-3 was down-regulated in OP group ( P<0.05). Compared with OP group, the apoptosis rate was significantly increased, the expression of Sigma-1R was down-regulated, and the expression of CHOP, p-IRE1, XBP1s and c-cas-3 was up-regulated in OPB group ( P<0.05). Conclusions:Sigma-1R is involved in pentazoxine-induced reduction of OGD/R injury in SH-SY5Y cells, and the mechanism may be related to inhibition of endoplasmic reticulum stress.

Objective:To investigate the clinical efficacy of 3D printed osteotomy guide plate combined with "Jail" screw technique in the treatment of tibial plateau fractures involving external posterior condylar collapse.Methods:From January 2016 to January 2021, 41 patients (22 males and 19 females) with tibial plateau fractures involving external posterior condylar collapse were treated with 3D printed osteotomy guide plate combined with "Jail" screw technique and followed up. The age was 47.4±11.5 years (range, 22-69 years). According to Schatzker fracture type, 18 cases were type IV, 14 cases were type V and 9 cases were type VI. All fractures were closed, and 12 of them were complicated with lateral meniscus injury, but none of them were complicated with nerve and vascular injury. The time from injury to operation was 7.2±3.4 d (range, 4-17 d). All patients underwent 3D CT scanning and digital modeling before operation. According to the modeling results, a 1∶1 solid size fracture model was made by 3D printing, and the osteotomy guide plate and the "Jail" screw preset guide plate were designed. During the operation, the tibial lateral condyle osteotomy was performed with customized osteotomy guide plate. After reduction, the fixation of the fracture was performed with the preset guide plate using "Jail" screw. Postoperative fracture reduction was evaluated according to Rasmussen score, and knee function was evaluated by Hospital for Special Surgery (HSS) score.Results:All the 41 patients were followed up for 15.2±5.8 months (range, 6-26 months). Immediate postoperative radiographs showed good fracture reduction, and the average healing time was 14.1±1.2 weeks (range, 12-17 weeks). One year after operation, the Rasmussen score of knee joint was 17.4±1.6 points (range, 13-19 points), of which 31 cases were excellent, 8 cases were good, and 2 cases were fair, with an excellent/good rate of 95% (39/41). HSS scores was 87.3±5.6 points (range, 68-95 points), including 30 excellent cases, 10 good cases and 1 fair case, with an excellent/good rate of 98% (40/41). The range of motion of knee joint was 126.8°±3.8°. At the last follow-up, no serious complications such as common peroneal nerve injury, popliteal vascular injury, postoperative infection, or internal fixation failure occurred.Conclusion:3D printed osteotomy guide plate combined with "Jail" nail placement technique is an effective method for tibial plateau fractures involving external posterior condylar collapse, and the postoperative treatment results are satisfactory. The use of customized osteotomy guide plate is more accurate and less damaging. The use of "Jail" screw preset guide plate can ensure more accurate screw placement.

Objective:To explore the practical role of massive open online course (MOOC) mode based on the cultivation of creativity and thinking ability in clinical human anatomy teaching.Methods:Two classes of clinical medicine students of Batch 2019 were selected in the study, and one class was control group ( n=73), which adopted the traditional teaching mode of face-to-face teaching; the other class was research group ( n=79) and the MOOC mode based on the cultivation of creativity and thinking ability for teaching was adopted. After the teaching, the students' creativity, thinking ability, self-learning ability and learning interest were compared, and the mastery of knowledge (theoretical scores and anatomical operation assessment) and satisfaction with teaching were compared between the two groups. SPSS 19.0 was used for t test, chi-square test and rank sum test. Results:The scores of creativity, thinking ability, self-study ability and learning interest of the research group were significantly higher than those of the control group after the teaching ( P<0.05). The scores of theoretical knowledge[(91.41±6.28) points] and anatomical operation[(87.41±7.25) points] in the research group were significantly higher than those in the control group after the teaching[(85.24±7.36) points and (80.26±6.38) points] ( P<0.05). There was significant difference in the distribution of teaching satisfaction between the two groups ( P<0.05), and the total satisfaction rate for teaching of the research group (94.94%) was higher than that of the control group (83.56%). Conclusion:The MOOC mode based on the cultivation of creativity and thinking ability in clinical human anatomy teaching can significantly improve students' creativity, thinking ability and self-learning ability, improve their learning interest and mastery of human anatomy knowledge, and improve their satisfaction with teaching.

OBJECTIVE: To explore the clinical features and genetic basis for a patient with hereditary hypophosphatemic rickets with hypercalciuria(HHRH). METHODS: Clinical data of the patient was collected. The patient was subjected to whole exome capture and next generation sequencing (NGS). Suspected variants were verified by Sanger sequencing. RESULTS: The patient presented with hypophosphatemic rickets, short stature, hypercalciuria, and renal stones. NGS showed that he has carried compound heterozygous variants of the SLC34A3 gene, namely c.532_533delCA(p.Q178Vfs*6) and c.894_925+69del(splicing). His parents were asymptomatic heterozygous carriers of one of the variants. Based on ACMG guidelines, both variants were classified as pathogenic. CONCLUSION The compound heterozygous variants c.532_533delCA (p.Q178Vfs*6) and c.894_925+69del(splicing) of the SLC34A3 gene probably underlie the disease in this child. Above finding has enriched the variant spectrum for HHRH. Based on the results, prenatal diagnosis may be provided for the family.

Objective: To retrospectively analyze the laboratory findings and ultrasonographic features in acute phase of children suffering from Kawasaki disease (KD) with stable hemodynamics and Kawasaki disease shock syndrome (KDSS), so as to provide the evidence for early diagnosis, timely treatment and improvement of prognosis of KDSS. Methods: Four hundred and eighteen patients with KD diagnosed at Shenzhen Children′s Hospital from November 2016 to May 2018 were selected, including 23 KDSS patients(KDSS group) and 395 cases with stable hemodynamic(KD without shock group). The clinical characteristics, laboratory index and ultrasonic examination data of the 2 groups were collected and compared for statistical conclusion. Results: (1)The level of C-reaction protein(CRP)[166.20 mg/L (74.40 mg/L)], γ-glutamyl transpeptidase(γ-GT)[88.00 IU/L (126.00 IU/L)], creatine kinase isoenzyme(CKI)[1.78 μg/L (5.17 μg/L)], troponin(TP)[0.01 μg/L (0.39 μg/L)] in the KDSS group in acute phase were all higher than those in the KD without shock group[70.50 mg/L (54.30 mg/L), 40.00 IU/L (89.00 IU/L), 1.20 μg/L (0.85 μg/L), 0.01 μg/L (0.01 μg/L)], hemoglobin(Hb)[90.00 g/L (15.00 g/L)], ablumin [24.20 g/L (4.30 g/L)], serum sodium[130.90 mmol/L (5.60 mmol/L)] levels in the KDSS group were lower than those in the KD without shock group[107.00 g/L (14.00 g/L), 33.40 g/L (4.08 g/L), 136.10 mmol/L (3.25 mmol/L)], and the differences were statistically significant (all P<0.05). (2)The incidence rates of impaired left ventricular ejection fraction(LVEF)[<55%: 3 cases (13.03%) vs.8 cases (2.00%)], coronary artery abnormalities[left anterior descending branch(LAD) Z-score>2.5: 6 cases (26.09%) vs.35 cases (8.86%)]and valvular regurgitation[tricuspid regurgitation(TR)≥moderate: 3 cases (13.03%) vs.5 cases (1.26%)]in the KDSS group were higher than those in the KD without shock group, and the differences were statistically significant (all P<0.05). (3)Among acute phase in KDSS group, 9 cases (39.13%) had liver enlargement, 9 cases (39.13%) had peritoneal effusion, 3 cases (13.04%) had diffuse renal lesions, 3 cases (13.04%) had joint effusion (2 cases of knee joint effusion, 1 case of hip joint effusion), and 2 cases (8.70%) had enteritis.In the KD without shock group, only 3 cases (0.76%) had hepatomegaly and 2 cases (0.51%) had a small amount of knee effusion. Conclusions Laboratory findings of KDSS group showed higher level of CRP, CKI, TP as well as γ-GT than those in KD without shock group, who are more prone to suffer from hypohemoglobin, hyponatremia and hypoalbuminemia.Ultrasound examination showed that KDSS children were more prone to have heart or multiple-organ damage.Clinicians should raise their awareness to provide comprehensive assessment as well as timely and effective treatment.

Objective: To investigate the mechanism of miR-31-5p/tension protein 1 gene (TNS1) axis modulating radiotherapy resistance in breast cancer. Methods: The breast cancer tissues and corresponding para-cancerous tissues of 21 patients with breast cancer, who underwent surgical resection at Department of Cancer Radiotherapy of Nanyang Central Hospital from July 2017 to December 2017, were collected for this study; breast cancer cell lines (MCF-7，MDA-MB-23 and SKBR-3) were also collected; qPCR was applied to detect the expression level of miR-31-5p in breast cancer tissues and cell lines. The radiation resistant cell line MCF-7R was constructed by using 6 MV-X ray radiotherapy treatment. Subsequently, the influence of over-expression/kockdown of miR-31-5p on radiation sensitivity of MCF-7 and MCF-7R cells were detected by colony formation assay, Transwell assay and Annexin V-FITC/PI double staining flow cytometry assay, respectively. Moreover, luciferase reporter assay was used to verify whether TNS1 was a target gene of miR-31-5p. Results: Compared with para-cancerous tissues, normal mammary epithelial MCF-10A cells and MCF-7 cells, miR-31-5p was low-expressed in breast cancer, cell lines and MCF-7R (all P<0.01). Over-expression of miR-31-5p resulted in inhibited invasion and promoted apoptosis of MCF-7R cells (P<0.01), whereas miR-31-5p knockdown got opposite results in MCF-7 cells. Moreover, luciferase reporter assay confirmed that TNS1 was a target gene of miR-31-5p. Over-expression of miR-31-5p inhibited invasion and increased radio-sensitivity, apoptosis of MCF-7R cell via targeting TNS1 (P<0.01), whereas knockdown of miR-31-5p significantly promoted the invasion but reduced apoptosis of MCF-7R cells (all P<0.01), and further up-regulated the radio-sensitivity of MCF-7R cells. Conclusion: miR-31-5p/TNS1 axis regulates the radiotherapy resistance of breast cancer, and over-expression of miR-31-5p may reverse the resistance of MCF-7R to radiotherapy.

OBJECTIVETo explore the clinical characteristics of a patient with Sensenbrenner syndrome (also called cranioectodermal dysplasia type 3) caused by mutation of intraflagellar transport (IFT) 43 gene.

METHODSThe clinical data of the patient was retrospectively analyzed. The target genes was the patient were captured and subjected to next generation sequencing. Suspected mutations were verified through Sanger sequencing.

RESULTSThe patient, a-13 year-and-5-month-old girl, was admitted for anemia and renal dysfunction for 8 months. Clinically, she has featured short stature, short limbs, brachydactylia, tooth agenesis, and retinal dystrophy, high-degree myopia, and chronic renal failure. Gene sequencing showed that she has carried a homozygous c.1A>G (p.M1V) mutation of the IFT43 gene, for which both of her parents were heterozygous carriers.

CONCLUSIONc.1A>G (p.M1V) mutation of the C14ORF179/IFT43 gene is the cause for praecox chronic renal failure in children. Genetic testing can facilitate the diagnosis of this rare disorder. For affected families, prenatal diagnosis should be provided.

Objective To evaluate effect of dexmedetomidine on mitochondrion-dependent apoptosis during hypoxia-reoxygenation (H/R) injury to hippocampal neurons of rats.Methods The primarily cultured hippocampal neurons of Sprague-Dawley rats were divided into 4 groups (n =40 each) using a random number table method:control group (C group),vehicle group (V group),H/R group and dexmedetomidine group (D group).Hippocampal neurons were subjected to oxygen-glucose deprivation followed by restoration of oxygen supply to establish the model of H/R injury.Dexmedetomidine 1 μmol/L was added at 6 h of reoxygenation in D group.The viability of neurons was measured by methyl thiazolyl tetrazolium assay at 20 h of reoxygenation.The ultrastructure of mitochondria was observed by transmission electron microscopy.The expression of cytochrome c (Cyt c),caspase-3,Fis1 and Drp1 was detected by Western blot.The neuronal apoptosis was detected by flow cytometry,and apoptosis rate was calculated.Results Compared with C group,no significant change was found in the viability of neurons in group V (P＞0.05),and the viability of neurons was significantly decreased,the apoptosis rate was increased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was up-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was accentuated in H/R and D groups.Compared with H/R group,the viability of neurons was significantly increased,the apoptosis rate was decreased,the expression of Cyt c,caspase-3,Fis1 and Drp1 was down-regulated (P＜0.05),and the damage to mitochondrial ultrastructure was significantly attenuated in D group.Conclusion The nechanism by which dexmedetomidine reduces the H/R injury to hippocampal neurons is related to inhibiting mitochondrion-dependent apoptosis in rats.