Objective To evaluate the effect and mechanism of terminal fucosylation inhibitor 2-deoxy-D-galactose (2-D-gal) on ciclosporin (CsA)-induced renal epithelial-mesenchymal transition (EMT). Methods Fifteen male C57BL/6 mice aged 8-10 weeks were randomly and evenly divided into the control group (Ctrl group), CsA group and CsA+2-D-gal group (n=5). The expression levels of fucosyltransferase 1 (FUT1), EMT-associated proteins including E-cadherin, Vimentin, α-smooth muscle actin (α-SMA) in the kidney tissues of the Ctrl and CsA groups were detected by Western blot. The expression levels of terminal fucose in the kidney tissues of Ctrl and CsA groups were determined by immunofluorescence. The renal fibrosis of mice in each group was evaluated by Masson staining. The blood urea nitrogen and serum creatinine levels of mice in each group were detected. The in vitro EMT model of renal tubular epithelial cell HK2 was induced by CsA. HK2 cells were stimulated with 0, 2.5, 5.0 and 10.0 μmol/L CsA for 24 h, respectively. In addition, HK2 cells were divided into the Ctrl, 2-D-gal, CsA and CsA+2-D-gal groups. The morphology of HK2 cells after stimulation with different concentrations of CsA and in each group was observed. The expression levels of FUT1, E-cadherin, Vimentin and α-SMA in HK2 cells after stimulation with different concentrations of CsA and in each group were detected by Western blot. The expression level of terminal fucose in HK2 cells of the Ctrl and CsA groups was measured by immunofluorescence. Results Compared with the Ctrl group, the relative expression of E-cadherin protein was down-regulated, those of FUT1, Vimentin and α-SMA proteins were up-regulated (all P < 0.05), and that of terminal fucose in the mouse kidney tissues was up-regulated in the CsA group. Compared with the Ctrl group, the blood urea nitrogen and serum creatinine levels in the CsA and CsA+2-D-gal groups were up-regulated (all P < 0.05). Compared with the CsA group, the blood urea nitrogen and serum creatinine levels in the CsA+2-D-gal group were down-regulated (both P < 0.05). Compared with the Ctrl group, the collagen fiber deposition was increased and the relative expression of α-SMA protein was up-regulated in the mouse kidney tissues of CsA and CsA+2-D-gal groups (all P < 0.05). Compared with the CsA group, the collagen fiber deposition was decreased and the relative expression of α-SMA protein in the mouse kidney tissues was down-regulated in the CsA+2-D-gal group (both P < 0.05). With the increase of CsA concentration, the morphology of HK2 cells gradually became longer and thinner from original normal cobblestone shape, the relative expression levels of FUT1, Vimentin and α-SMA protein in HK2 cells were up-regulated, and that of E-cadherin protein was down-regulated in a concentration-dependent manner. Compared with the Ctrl group, the expression level of terminal fucose of HK2 cells was up-regulated in the CsA group. After CsA treatment combined with 2-D-gal intervention, the morphology of HK2 cells in the CsA+2-D-gal group was restored to resemble that of normal HK2 cells. Compared with the CsA group, the relative expression of E-cadherin protein in HK2 cells was up-regulated, whereas those of Vimentin and α-SMA proteins were down-regulated in the CsA+2-D-gal group (all P < 0.05). Conclusions CsA may induce EMT both in vivo and in vitro, and the terminal fucosylation is increased. 2-D-gal may inhibit CsA-induced EMT by suppressing the terminal fucosylation.

BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.

ObjectiveTo investigate the effect of the Yap1 gene and tanshinone ⅡA on the proliferation, migration, and invasion abilities of Huh-7 hepatoma cells. MethodsA total of 10 pairs of human hepatocellular carcinoma (HCC) samples and adjacent tissue samples were collected in Zhongnan Hospital of Wuhan University from June 1 to December 1, 2019. Quantitative real-time PCR and Western blotting were used to measure the expression of the Yap1 gene and phenotype-related molecules. MTT cell proliferation detection reagent was used to measure the inhibition rate of cell proliferation after the treatment with different concentrations of tanshinone ⅡA. Western blotting was used to measure the changes in the expression of apoptosis-and migration-related markers after different interventions. Flow cytometry and Transwell assay were used to measure apoptosis and cell migration and invasion abilities. The data of 375 cases of liver cancer and 50 cases of relatively normal liver tissue samples were downloaded from The Cancer Genome Atlas, including clinicopathological information. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. ResultsIn 8 of the 10 pairs of HCC samples and adjacent tissue samples, HCC samples had significantly higher expression of Yap1 than the adjacent tissue samples. Compared with the normal human liver epithelial cells L02, the Huh-7 and HCCL-M3 hepatoma cells had a significant increase in the expression of Yap1. The silencing efficiency of si-Yap1-3 transfection reached 87.004% at the protein level. MTT results showed that tanshinone ⅡA effectively inhibited the proliferation of Huh-7 cells, with a half inhibitory concentration of 8.683 μmol/L. After the cells were treated with si-Yap1-3 and tanshinone ⅡA, there was an increase in the expression of the downstream marker for proliferation and migration E-cadherin and a reduction in the expression of vimentin, and the results of Transwell assay showed that compared with the si-NC group, the tanshinone ⅡA+si-Yap1-3 group had significant reductions in the migration and invasion abilities of Huh-7 cells (migration: 43.19±2.88 vs 132.20±10.03, t=8.527, P=0.001; invasion: 53.95±4.20 vs 179.10±11.11, t=4.484, P=0.011). The group treated with si-Yap1-3 and tanshinone ⅡA had an increase in the expression of the apoptosis-related marker Bax and a reduction in the expression of Bcl-2, as well as a significantly higher early apoptosis rate than the si-NC group (2598% vs 9.21%, χ2=4.078, P＜0.05). ConclusionOncogene Yap1 silencing combined with tanshinone ⅡA can promote the apoptosis of Huh-7 hepatoma cells and inhibit their migration and invasion, which can provide certain guiding significance for clinical medication.

Objective To analyze the epidemiological features of Galeazzi fractures from 2003 through 2012 in The Third Affiliated Hospital to Hebei Medical University. Methods We retrospectively analyzed the data of Galeazzi fractures between January 2003 and December 2012 in the Third Affiliated Hospital to Hebei Medical University. The data from 2003 through 2008 were defined as group A and those from 2008 through 2012 as group B. The general information was compared between the 2 groups. The epi-demiological characteristics and trends during the 10 years were analyzed concerning gender, age and fracture type of the patients. Results A total of 153 Galeazzi fractures were recorded, accounting for 0. 81％ of ulnoradial fractures and 0. 12％ of all fractures. There were 109 males and 44 females, with a male/female ratio of 2. 48:1. The age range from 11 to 20 years had the highest constituent ratio ( 22. 22％) and type Ⅱthe highest proportion ( 76. 47％) . There were 74 cases in group A, with a male/female ratio of 2. 22:1. There were 79 cases in group B, with a male/female ratio of 2. 76:1. There was no significant difference between the 2 groups concerning the male/female ratio ( P > 0. 05 ) . The median age for group A was 29 years, significantly younger than that for group B ( 34 years ) ( P <0. 05 ) . The high risk age was from 11 to 20 years in group A (32. 43％) and from 21 to 50 years (22. 78％) in group B. Compared with group A, the constituent ratio of age range from 11 to 20 years in group B was significantly lower and the constituent ratio of age range from 41 to 50 years significantly higher ( P <0. 05 ) . There were no significantly differences between the 2 groups concerning the constituent ratio of each fracture type ( P> 0. 05 ) . Conclusions Galeazzi fractures accounted for 0. 81％ of ulnoradial fractures and 0. 12％ of all fractures. There were more male patients than female ones. The age range from 11 to 20 years and typeⅡhad the highest constituent ratios. Compared with the first 5 years, the latter 5 years witnessed increased mean age.

Objective To compare and analyze the epidemiological features of adult patients with scapular fracture between eastern and western areas in China from 2010 to 2011. Methods The data of adult patients with scapular fracture who had been treated from January 2010 to December 2011 in 63 hospitals in the eastern and western areas in China were collected through the PACS system and case reports checking system. The data from the eastern 35 hospitals were classified as group A and those from the 28 western hospitals as group B. The analytic items included gender, age and AO classification. Results A total of 2, 063 cases were collected. The median age was 44 years ( interquartile range, from 33 to 56 years ) and gender ratio 3. 50:1 in Group A of 1, 376 cases; the median age was 41 years ( interquartile range, from 31 to 51 years ) and gender ratio 3. 09:1 in Group B of 687 cases. There was a significant difference between the 2 groups in median age ( Z= -3. 798, P=0. 000 ) . There was no significant difference between the 2 groups in gender ratio or constituent ratio of fracture peak age ( P > 0. 05 ) . The proportions of patients from 16 to 20 and from 31 to 40 years old in group A were significantly lower than in group B ( P <0. 05 ) . The proportion of middle-aged patients was significantly higher and that of young patients significantly lower in group A than in group B. The proportion of AO type 14-A fractures in group A ( 66. 49％) was significantly higher than in group B ( 55. 60％) while the proportion of AO type 14-C fractures was significantly lower ( 21. 37％) in group A than in group B ( 29. 55％) . Conclusions Most scapular fractures occurred in male patients. The proportion of young patients in the eastern area was lower than in the western area, but the proportion of middle-aged patients was higher in the eastern area than in the western area. The proportion of AO type 14-A fractures in the eastern area was significantly higher than in the western area while the proportion of AO type 14-C fractures in the eastern area significantly lower than in the western area.

Objective To compare and analyze the epidemiological features of adult patients with distal humerus fractures between the east and west China from 2010 to 2011.Methods The data of adult patients with distal humerus fracture treated from January 2010 to December 2011 in 63 hospitals in the east and west China were collected through the PACS system and case reports checking system.The data from 35 hospitals in the east China were classified as group A,and those from 28 hospitals in the west China as group B.The analytic items included gender,age and AO/OTA classification.Results A total of 2,668 cases were collected.The median age of the 1,852 cases in group A was 42 (IQR:33) years old.The high peak age ranged from 21 to 30 years old;the gender ratio of group A was 1.18:1.The median age of the 816 cases in group B was 37 (IQR:27) years old.The high peak age ranged from 31 to 40 years old;the gender ratio of group B was 1.80:1.Group A had a significantly lower gender ratio but a significantly higher median age than group B (P ＜ 0.05).The proportions of patients from 16 to 20 and from 31 to 40 years old in group A were significantly lower than those in group B but the proportion of patients aged above 61 years old in group A was significantly higher than that in group B (P ＜ 0.05).According to the AO/OTA classification,1,399 cases (52.44％) were type 13-A,682 ones (25.56％) type 13-B and 587 ones (22.00％) type 13-C.The proportions of type 13-A and type 13-A1 in group A were significantly lower than in group B (P ＜ 0.05).Conclusions Fractures of the distal humerus mostly occurred in the age range from 21 to 40 years old and in males.Type 13-A fractures were the most common.The east China had a significantly higher proportion of female patients,an older median age and a lower proportion of type 13-A1 fracture than the west China.

Objective To compare the epidemiological characteristics of adult single fractures of the cuneiform between the east and west areas in China from 2010 through 2011.Methods Thirty-five hospitals in the east coast area (group A) and 28 hospitals in the west inland area (group B) were selected for this investigation. The data of adult single fracture of the cuneiform treated between January 2010 and De-cember 2011 at the 63 hospitals were collected through the PACS system and medical records inquiry system. The epidemiological characteristics concerning age, gender and fracture classification were compared between the 2 groups. Results A total of 457 adult single fractures of the cuneiform were collected, involving 384 cases in group A and 73 cases in group B. They accounted respectively for 4.38% (384/8,772) and 3.53% (73/2, 068) of the adult tarsal bone fractures in the same period, showing no significant difference (P >0.05), accounted respectively for 1.71% (384/22, 455) and 1.15% (73/6, 354) of the adult foot frac-tures, showing a significant difference (P <0.05), and accounted respectively for 0.20% (384/192,991) and 0.09% (73/81, 143) of all the adult fractures, showing a significant difference (P <0.05). The median age was 39 years in group A (29, 50) and 36 years in group B (25, 45), showing a significant difference (P <0.05). The male to female ratio was 1.61:1 (237/147) for group A and 1.52:1 (44/29) for group B. The peak age range of the fracture for males was from 21 to 30 years old in both groups, and its proportion was 28.69% (68/237) in group A and 38.64% (17/44) in group B. The peak age range of the fracture for females was from 41 to 50 years old in both groups, and its proportion was 26.53% (39/147)in group A and 27.59% (8/29) in group B. All the above comparisons were statistically insignificant (P> 0.05). The proportion of the age group of≥61 years was significantly larger in group A than in group B (P <0.05). The high risk type of the fracture was type 85-A1 in both groups, and its proportion was 61.98% (238/384) in group A and 52.05% (38/73) in group B. The proportion of type 85-B1 was significantly larger in group B than in group A (P <0.05). Conclusions Adult single fractures of the cuneiform were more common in young and male patients. The proportion of elderly patients (≥61 years old) was significantly larger in the east area. The high risk type of the fracture was simple fracture involving the medial cuneiform bone (type 85-A) in both areas. The proportion of the comminuted fracture of the medial cuneiform (type 85-B1) was significantly larger in the west area.