Juglone induces proliferation inhibition and apoptosis of cervical cancer cells via promoting c-Myc ubiquitination / 南方医科大学学报

OBJECTIVE@#To observe the expression of c-Myc protein in cervical cancer HeLa cells and explore the effect of juglone on the proliferation and apoptosis of HeLa cells by affecting c-Myc ubiquitination.@*METHODS@#HeLa cells treated with different concentrations (0, 10, 20, or 50 μmol/L) of juglone or with 20 μmol/L juglone for different time lengths were examined for expression of c-Myc protein with Western blotting. The half-life of c-Myc protein was determined using cycloheximide (CHX) and c-Myc protein degradation was detected using coimmunoprecipitation. We also assessed the effects of 20 μmol/L juglone combined with 0, 1.0 or 2.0 μmol/L MG132 (a proteasome inhibitor) on c-Myc expression. The effects of 20 μmol/L juglone on the proliferation and apoptosis of HeLa cells with RNA interference-mediated knockdown of c-Myc were evaluated with MTT assay and flow cytometry.@*RESULTS@#Treatment with juglone significantly lowered c-Myc protein expression in HeLa cells in a concentration-and time-dependent manner (P < 0.05). Juglone obviously shortened the half-life of c-Myc protein, and the addition of MG132 significantly up-regulated the expression level of c-Myc protein (P < 0.05). Juglone treatment also promoted ubiquitination of c-Myc protein in HeLa cells. Compared with the cells transfected with a negative control construct, the cells transfected with si-c-Myc showed significantly decreased proliferation inhibition and a lowered cell rate with early apoptosis after juglone treatment (P < 0.05).@*CONCLUSION@#Juglone inhibits proliferation and promotes apoptosis of HeLa cells by affecting the ubiquitination of c-Myc protein.

Effect and mechanism of 6-gingerol on invasion and migration of HPV-positive and negative cervical cancer cells / 解放军医学杂志

Objective To observe the inhibitory effect of 6-gingerol on the invasion and migration of human papilloma virus (HPV)-positive and HPV-negative cervical cancer cells, and explore the possible mechanism. Methods Human HPV-positive cervical cancer cells (HeLa) and HPV-negative cells (C33A) were cultured, and added with 6-gingerol at different concentration of 0, 5, 10, 20, 50 μmol/L, the untreated cells play as control, then cultured for 24 h. 10 μmol/L 6-gingerol was determined as the best concentration, then all the cells were treated with 10 μmol/L 6-gingerol for 24 h, 48 h and 72 h respectively. The cell proliferation was detected by MTS method, and cell scratch test was performed to detect the effect of 6-gingerol on cell migration ability. The effect of 6-gingerol on cellular invasion was detected by Transwell chamber. Western blotting was used to detect the expression of matrix metalloproteinase MMP-2, MMP-9, E-cadherin and N-cadherin. Results MTS assay showed that the activity of HeLa and C33A cells decreased with the increase of 6-gingerol concentration and action time, while the activity of HeLa cells decreased more significantly than that of C33A cells at different concentrations or time points. Transwell invasion chamber test showed that the HeLa cells treated with 6-gingerol for 24 h and 48 h, and C33A cells treated with 6-gingerol for 48 h, the cellular invasion ability decreased significantly. The scratch test revealed that the wound healing rate decreased significantly of HeLa cells 24 h and 48 h after 6-carbenol action and of C33A cells 48 h after 6-carbenol action. Western blotting results showed that, treating with 10 μmol/L of 6-gingerol for 24 h, the expressions of E-cadherin increased and of N-cadherin, MM P-2 and MMP-9 declined in HeLa cells with statistical differences (P0.05) compared to that of unprocessed group. Conclusions 6-gingerol can inhibit the proliferation of HPV-positive cervical cancer cells and cell invasion. The mechanism may be associated with the effect of 6-gingerol on influencing the expression of epithelial-mesenchymal transition-related proteins.

Long-term outcomes of Chiari osteotomy for Legg-Calvé-Perthes disease in children / 中国骨伤

OBJECTIVE@#To explore long-term outcomes of Chiari osteotomy for Legg-Calvé-Perthes disease in children with type Catterall III or IV, and to analyze clinical effect of osteotomy angle on clinical and radiographic results.@*METHODS@#From March 2005 to July 2013, 26 children with Legg-Calvé-Perthes disease with type Catterall III or IV were treated by Chiari osteotomy, including 17 males and 9 females, aged from 4 to 13 years old with an average of (8.9±2.6) years old. Children were divided into low osteotomy angle group and high osteotomy angle group. according to osteotomy angle. There were 10 children in low osteotomy angle group with an osteotomy angle of 10 degrees, including 8 boys and 2 girls, aged from 4 to 13 years old with an average of (9.2±3.3) years old; while there were 16 children in high osteotomy angle group with an osteotomy angle of 15 degress, including 9 boys and 7 girls, aged from 6 to 12 years old with an average of (8.8±2.1) years old. HHS score before operation and at the latest follow-up were recorded to observe clinical results. CE angle of hip joint, acetabular index, Sharp angle, Shenton's line continuity, femoral head coverage, acetabular depth ratio were recorded to compare radiographic results. Stulberg classification was analyzed to compare reshaping ability of femoral head.@*RESULTS@#Twenty-six children were followed up for 4.5 to 12.0 years with an average of (7.9±1.8) years. All incisions were healed at stage I for 10 to 14 days, with an average of(12.3±1.1) days. No inflammation, skin necrosis and injury of vessel and nerve occurred. All osteotomies achieved bone union for 8 to 13 weeks, with an average of(9.8±1.4) weeks. HHS score increased from 75.8±6.5 before operation to 93.5±2.5 at the latest follow-up in low osteotomy angle group(<0.05), and form 77.6±6.2 to 97.8±1.6 in high osteotomy angle group (<0.05). HHS score of high osteotomy angle group at the latest follow-up was higher than that of low osteotomy angle group (<0.05). The acetabular index decreased from (10.1±2.5)° before operation to (4.5±1.3)° at the latest follow-up in low osteotomy angle group (<0.05), and from (10.7±3.3)° before operation to (2.0±1.1)° in high osteotomy angle group (<0.05). The acetabular index of high osteotomy angle group at the latest followup was better than low osteotomy angle group(<0.05). There was no significant difference in CE angle, Sharp angle, Shenton's continuity, femoral head coverage, acetabular depth ratio between two groups. According to Stulberg classification, the femoral head reshaping ability in high osteotomy angle group was better than that of low osteotomy angle group(<0.05).@*CONCLUSIONS@#Chiari osteotomy with 15° for Legg-Calvé-Perthes disease in children with type Catterall III or IV could effectively decrease index of acetabulum, and helpful for femoral head reshaping ability, then in further improve clinical effects.

Clinical Characteristics of Brucellosis Complicated by Syndrome of Inappropriate Secretion of Antidiuresis / 中国医学科学院学报

To summarize and analyze the causes of hyponatremia in patients with brucellosis and explore the clinical manifestations of syndrome of inappropriate antidiuresis(SIAD)in patients with brucellosis. The clinical data of 111 patients with acute brucellosis who were treated in Peking Union Medical College Hospital from September 2011 to December 2017 were retrospectively reviewed.Hyponatremia was defined by serum sodium level lower than 135 mmol/L.Clinical characteristics including medical histories,vital signs,and laboratory test findings were collected and analyzed. Hyponatremia was found in 14(12.6%)of 111 patients with brucellosis,among whom 3 patients were confirmed to be with SIAD,10 were suspected as SIAD,and 1 was diagnosis as hypopituitarism.Hypoalbuminemia,elevation of erythrocyte sedimentation rate,and high sensitivity C reactive protein were found in brucellosis patients with SIAD,along with severe complications such as infective endocarditis,septic shock,and anemia. Hyponatremia is not a rare condition in brucellosis patients and may be caused by SIAD.

Repairing diabetic rats with bone defect by VEGF165 gene modified adipose-derived stem cells / 中国骨伤

<p><b>OBJECTIVE</b>To explore repairing results of VEGF165 gene modified adipose-derived stem cells for diabetic rats with bone defect.</p><p><b>METHODS</b>Seventy-eight male Wistar rats weighted 180 to 220 g were selected, 72 rats were established diabetic animal models by streptozotocin inducement method, blood glucose level was more than 16.7 mmol/L. Experimental animals were randomly divided into 5 groups, 6 rats in normal group and each 18 rats in other groups. VEGF165 gene modified adipose-derived stem cells were implanted into normal group with bone defect; single diabetic rats with bone defect were named as diabetic group;vascular endothelial growth factor implanted into single diabetic rats with bone defect named as growth factor group; adipose-derived stem cells implanted into diabetic rats with bone defect names as stem cell group; VEGF165 gene modified adipose-derived stem cells implanted diabetic rats with bone defect named as experimental group. After combination of VEGF165-ADSCs (5×106) cells combined with gel sponge, implanted into diabetic rats with bone defect. On the forth week, general form of defect repairing tissue were observed by optical microscopy;local density of micro-vessel were detected by immunohistochemistry method; content of Ca, P and ALP of repairing callus were detected by IRIS Intrepid XSP inductively coupled plasma emission spectrometer. Efficacy of the VEGF165-ADSCs repairing function was evaluated by SPSS statistic software.</p><p><b>RESULTS</b>Fluorescent staining results showed that expression of VEGF165 located on cytoplasm of ADSCs, expression percentage was more than 87%; general histology results showed that callus formation and quality was near to normal group, repairing results in diabetes group, growth factor group and stem cell group were poor. On the Forth week after implantation, content of Ca, P and ALP of repairing callus in experimental group were higher than those in growth group and stem cell group, and without significant differences compared with normal group; blood vessel density in experimental group was lower than normal group, but higher than other groups.</p><p><b>CONCLUSIONS</b>VEGF165 gene modified adipose-derived stem cells for repairing diabetic rats with bone defect has advantages of osteogenesis and angiogenesis, and should be one of the effective method for repairing diabetic rats with bone defect.</p>

Effect and its mechanism of Juglone on invasion and metastasis ability of human cervical cell line Hela / 解剖学报

Objective: To observe the effect of Juglone on invasion and metastasis of Hela cells and explore the possible mechanism. Methods: HeLa cells were cultured and treated with 10,20,50,100μmol/L Juglone for 24 hours. The morphology changes of HeLa cells were observed with an inverted microscope. The viability of HeLa cells was detected by MTT assay. The cell scratch test was used to detect cell migration ability after treatment of Juglone. The ability of cell invasion was measured by Transwell chamber. The expression of matrix metalloprateinases (MMP)-2 and MMP-9 were detected by Western blotting. Results: Compared with control group, the viability of HeLa cells decreased after treatment with different concentrations of Juglone for 24 hours, and the cell morphology was changed in a dose-dependent manner. Scratch test results showed that the level of cell movement ability decreased significantly with the increase of the concentration of Juglone. Transwell invasion assay showed that Juglone had a strong inhibitory effect on the invasiveness of HeLa cells in vitro. Western blotting results showed that Juglone inhibited the expression of MMP-2 and MMP-9 protein in HeLa cells. Conclusion: Juglone can inhibit the invasion and metastasis in HeLa cells, and its possible mechanism may be related to down regulating the expression of MMP-2 and MMP-9.