Objective To evaluate the outcomes of open reduction and internal fixation of complex tibial plateau fractures with improved three combined approaches.Methods In the period from July 2014 to February 2016,7 complex tibial plateau fractures underwent surgical treatment.These patients included 5 male and 2 female,aged from 24 to 68 years old (average,39.7± 15.3 years).According to Schatzker classification,they were all of type V.And all of type 41B-3.1 by AO/OTA classification.All these fractures were exposed and reduction via three combined approaches.First let the patients lied in lateral prone position,expose the anterolateral and the posterolateral of the tibia1 plateau,and fix the fractures of the posterior in the posterolateral approach,then fix the fractures of the lateral in the anterolateral approach.Then turn the patients to supine position,fix the fractures of the medial in the anteromedial incision.All patients received regular reexamination.The knee function was evaluated at the final follow-up using The Hospital for Special Surgery (HSS) score,the activity of the knee was evaluated by Lysholm score,and the stability of the knee was checked by Lachmantest and Pivot-shift test.The tibial plateau angle,the posterior slope angle and Rasmussen X-ray score were assessed on the X-ray films.Results The average time of operation is (3.3±0.9) h,and the hemorrhage volume in operation was (341± 106) ml for the 7 patients.The wounds of 6 patients healed by (11.8± 1.3) days,while the wound of the rest one of them occurred fat liquefaction after operation who is very fat (BMI ＞ 30) and suffered from diabetes.His wound healed by 21 days.These patients obtained follow-ups of 8 to 14 months (average,11.4±2.8 months).The average full weight-bearing time was 2-3 months (average,2.5±0.4 months).The fractures healed after 8 to 16 weeks (average,11.1 ±2.8 weeks).No displacement of the fractures or breakage of the implants occurred in our series.Nobody has activity limitation of the knee or pain because of the implant,and no never symptoms were noted postoperatively,so we would not take out the implant for these patients.The mean HSS score was 93.1±4.8 (range,from 85 to 100) at the final follow-up,and the excellent rate is 100％.The Lysholm score was 97.1±3.6 (range,from 90 to 100) at the final follow-up.The Lachman test and the Pivot-shift test were negative in our patients,and the mean knee flexion was 128.6±12.8°(range,from 105°to 140°).The fractures were all anatomical reduction by the X-ray after operation,and there had being no displacement of the fractures or breakage of the implants occurred during the follow-up period.The mean posterior slope angle was 8.29±2.87° (range,from 4 °to 12°),which was 8.71±2.63° (range,from 5 °to 14°) 6months after operation.The mean tibial plateau angle was 86.00± 1.41° (range,from 84 °to 88°),which was 86.43± 1.62° (range,from 84 °to 89°) 6 months after operation.The mean Rasmussen X-ray score was 16.86±1.57 (range,from 14 to 18),which was 16.57±1.51 (range,from 14 to 18) 6 months after operation,and the excellent rate are 100％.Conclusion For the complex tibial plateau fractures which simultaneous involved the medial,the lateral and the posterior,the improved three combined approaches showed the advantages of the convenient operation,the satisfactory results of reduction and fixation,and the less trauma and secondary damage,and could be worth for clinic.

Objective To investigate the therapeutic mechanism of bazedoxifene, the third-generation selective ER modulator (SERM), on endometriosis lesions in a rat model. Methods Endometriosis was induced by transplanting pieces of endometrium from other syngeneic rats that were as donors onto the subcutaneous of other unmated female rats. The rats with successful ectopic implants were divided into two groups:control group (n=10) and bazedoxifene group (n=10). The macroscopic morphology, volume, histopathology of ectopic implant and rats uterine wet weight were determined before and after the treatment. Expression of proliferation cell nuclear antigen (PCNA), ER and PR in the eutopic endometrium and endometriosis lesions detected by immunohistochemistry in the two groups. Results (1) The gross morphology and histological changes of endometriosis lesions in rats after treatment: compared with the control group, it was obviously depauperated and had more less glands and blood vessels in the stroma. (2) The change of rats′weight, the volume of endometriosis lesion before and after treatment and rats uterine wet weigh after treatment respectively in the control group and the bazedoxifene group:rats′ weight were respectively before treatment: (201±17) g, (202±18) g, that were respectively after treatment: (266±16) g, (261±16) g, which showed no significant difference between two groups before and after treatment (P>0.05). The volume of ectopic implant before treatment were respectively (85±17) mm3, (85±12) mm3, and showed no significant difference between two groups;that were respectively (48±11) mm3, (24±9) mm3 afte rtreatment, which was significantly decreased compared with the control group (P<0.05). Rats uterine wet weight after treatment were respectively:(0.77±0.16) g, (0.45±0.18) g, and was significantly reduced compared with the control group (P<0.05). (3) The protein expression levels of PCNA, ER and PR in the endometriosis lesions after treatment were respectively 0.282 ± 0.044, 0.51 ± 0.06, 0.49 ± 0.05 in the control group, 0.191 ± 0.020, 0.23 ± 0.03, 0.48 ± 0.06 in the bazedoxifene group; that in eutopic endometrium were respectively 0.369 ± 0.081, 0.56 ± 0.08, 0.56 ± 0.10 in the control group, 0.211 ± 0.037, 0.27 ± 0.05, 0.54 ± 0.08 in the bazedoxifene group;the protein expression levels of PCNA and ER in endometriosis lesions and the eutopic endometrium were significantly decreased in the bazedoxifene group compared with the control group (P<0.05), but the protein levels of PR in endometriosis lesionsand and the eutopic endometrium were not significantly altered by treatment (P>0.05). Conclusion Bazedoxifene could obviously reduce the size of endometriosis lesions, the mechanism may be related with suppressing estrogen-induced proliferation, the expression of ER and direct ER antagonism by this SERM.

Obj ective To observe the position fertilization (IVF)treatment cycle of in vitro cultivation quality blastocyst formation and analyze its relationship with pieces of D2D3 embryo cleav-age count and rating.Methods Cleavage stage 3 day (D3 )frozen surplus embryos were selected and cultured to blastocyst stage.Quality of blastocyst formation was observed,and at the same time their D2D3 embryo cleavage count and debris were discussed.Results The high quality D3 10 or more cells blastocyst formation rate was 38.9%,which was almost equal to 43.1%of 7 to 9 cells blastocyst formation rate (P>0 .05 ).7 to 9 cell quality blastocyst formation rate was 43 .1%, which was significantly higher than 10.2%of 4 to 6 cells (P<0.05).D2 4 cell quality blastocyst formation rate was 43.1%,which was significantly higher than 28.9%of 5 cells and 12.3%of 2 to 3 cells (P<0 .05 ).5 cells quality blastocyst formation rate was 28 .9%,which was almost equal to 23 .4%of 6 cells blastocyst formation rate (P>0 .05 ).Ⅰand classⅡD3 embryos pieces of high quality blastocyst formation rate was 36.4%,which was significantly higher than 6.1%of gradeⅢorⅣ(P<0 .05 ).Ⅰand class ⅡD2 embryos pieces of high quality blastocyst formation rate was 35.3%,which was significantly higher than 4.7% of grade Ⅲ or Ⅳ (P<0.05).Conclusion Extending IVF can effectively filter the embryo developmental potential.The more D3 embryo cleavage count ,the more high-quality blastocysts .D2 4 cells ,embryo blastocyst development ability is good.Ⅰand classⅡD2 and D3 embryos pieces of high quality blastocyst formation rate is higher.High quality blastocyst formation and D2 and D3 embryo cleavage count and debris are closely related to the ratings.

Obj ective To observe the position fertilization (IVF)treatment cycle of in vitro cultivation quality blastocyst formation and analyze its relationship with pieces of D2D3 embryo cleav-age count and rating.Methods Cleavage stage 3 day (D3 )frozen surplus embryos were selected and cultured to blastocyst stage.Quality of blastocyst formation was observed,and at the same time their D2D3 embryo cleavage count and debris were discussed.Results The high quality D3 10 or more cells blastocyst formation rate was 38.9%,which was almost equal to 43.1%of 7 to 9 cells blastocyst formation rate (P>0 .05 ).7 to 9 cell quality blastocyst formation rate was 43 .1%, which was significantly higher than 10.2%of 4 to 6 cells (P<0.05).D2 4 cell quality blastocyst formation rate was 43.1%,which was significantly higher than 28.9%of 5 cells and 12.3%of 2 to 3 cells (P<0 .05 ).5 cells quality blastocyst formation rate was 28 .9%,which was almost equal to 23 .4%of 6 cells blastocyst formation rate (P>0 .05 ).Ⅰand classⅡD3 embryos pieces of high quality blastocyst formation rate was 36.4%,which was significantly higher than 6.1%of gradeⅢorⅣ(P<0 .05 ).Ⅰand class ⅡD2 embryos pieces of high quality blastocyst formation rate was 35.3%,which was significantly higher than 4.7% of grade Ⅲ or Ⅳ (P<0.05).Conclusion Extending IVF can effectively filter the embryo developmental potential.The more D3 embryo cleavage count ,the more high-quality blastocysts .D2 4 cells ,embryo blastocyst development ability is good.Ⅰand classⅡD2 and D3 embryos pieces of high quality blastocyst formation rate is higher.High quality blastocyst formation and D2 and D3 embryo cleavage count and debris are closely related to the ratings.

Chronic endometritis (CE) is one of the common diseases in women of reproductive age, belonging to pelvic inflammatory disease, and characterized by a persistent localized inflammatory state of the endometrium. Clinically, CE often presents as asymptomatic or with atypical symptoms, leading to frequent neglect by obstetricians and gynecologists. In recent years, the incidence of CE has been increasing annually and has become a significant cause of unexplained infertility, recurrent implantation failure, and miscarriage in women. It also plays a crucial role in influencing the outcomes of assisted reproductive technologies. Therefore, safe, effective, and non-invasive diagnostic and therapeutic methods have garnered increasing attention. This article comprehensively elaborated on the etiology, latest diagnostic methods, and multidimensional treatment modalities of CE, providing novel insights into its diagnosis and treatment.

Acute traumatic spinal cord injury (SCI) represents one of the most devastating injuries that afflict the human body. Ascorbic acid ( AA) plays an important role in mammalian central nervous system, especially in SCI. In this study, the change of AA concentration after SCI was investigated by using an on-line electrochemical method integrated with in vivo microdialysis. A microdialysis probe (2 mm in length) was implanted into the spinal cord of an anesthetized rat (Thoracic-10). Microdialysis perfusate (2 μL/ min) was collected in the sample loop of an on-line injector for direct injection onto a glassy carbon electrode which was modified with the heat-treated single-walled carbon nanotubes (SWNTs). Normal ascorbic acid concentration in the extracellular fluids of spinal cords was (26. 17 ± 1. 25) μmol/ L (n =8). The experimental spinal cord injury, induced by a lesion at T-10, significantly increased the extracellular ascorbic acid levels to (53. 24± 1. 95) μmol/ L (n =8). This study provides the experimental evidence on the essential roles of ascorbic acid in spinal cord injuries.

AIM:To verify the safety application of MIL60 in the treatment of corneal neovascularization both in vivo and in vitro.METHODS:We observed the biological characteristics of human corneal epithelial cells.The cell proliferation was analyzed using CCK-8 assay,which also used to test the toxicity of MIL60 and the solvent on cultured human corneal epithelial (HCE).FACs was used to analyze the apoptosis of HCE after treated with MIL60.Also we evaluated the effect of subconjunctival injection of MIL60 on corneal epithelial healing model in normal rat and rats with epithelium defect through slit lamp-microscopy,Draize scores and histopathology way.RESULTS:The proliferation speed of HCE in three groups was the same.MIL60 did no harm on the proliferation of HCE and the apoptosis of HCE,and has no effect on corneal epithelial healing and other parts of the ocular in rats without inflammation cells infiltration.CONCLUSION:When given subconjunctival injection,Mil60 does no harm to the proliferation and apoptosis of HCE,and is safe with ocular application.

BACKGROUNDAtopic dermatitis (AD) is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 (Th1/Th2) cytokine expression. Filaggrin (FLG) is the key protein to maintaining skin barrier function. Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes. However, the role of Th1/Th2 cytokines on FLG processing is not substantially documented. Our aim was to investigate the impact of Th1/Th2 cytokines on FLG processing.

METHODSHaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin (IL)-4, 13 or interferon-γ (IFN-γ). FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor (LEKTI) were measured by both real-time quantitative polymerase chain reaction and Western blotting. Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.

RESULTSIL-4/13 significantly reduced, while IFN-γ significantly up-regulated FLG expression. IL-4/13 significantly increased, whereas IFN-γ significantly decreased the expression of kallikreins 5 and 7, matriptase and channel-activating serine protease 1. On the contrary, IL-4/13 significantly decreased, while IFN-γ increased the expression of LEKTI and caspase-14. Similar trends were observed in AD lesions.

CONCLUSIONSOur results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes. The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.

Caspase 14 ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Dermatitis, Atopic ; metabolism ; Humans ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Intermediate Filament Proteins ; metabolism ; Keratinocytes ; enzymology ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; metabolism ; Serine Peptidase Inhibitor Kazal-Type 5 ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

Objective:To compare the clinical efficacy between coracoclavicular ligament anatomical reconstruction and clavicular hook plate in the treatment of Neer Ⅱb distal clavicular fractures.Methods:A total of 64 patients with Neer Ⅱb clavicular fracture were treated at Department of Orthopaedics, The First Affiliated Hospital to Nanjing Medical University from September 2016 to June 2019. They were 35 males and 29 females, aged from 19 to 68 years (average, 50.7 years). They were assigned into 2 groups according to their operative methods: a reconstruction group of 30 cases undergoing coracoclavicular ligament anatomical reconstruction and a hook plate group of 34 cases undergoing fixation with a clavicular hook plate. The 2 groups were compared in terms of hospital stay, operation time, intraoperative blood loss, surgical incision length, postoperative coracoclavicular separation ratio, visual analogue scale (VAS) and Constant-Murley shoulder scores at 3, 6 and 12 months after operation, and postoperative complications.Results:There was no significant difference in general data between the 2 groups, showing comparability between groups ( P>0.05). Operations were completed uneventfully and surgical incisions healed by primary intention in both groups after operation. All the patients were followed up for 12 to 24 months (average, 14.6 months). The operation time [(74.6±22.0) min] and incision length [(10.4±0.4) cm] were significantly shorter but the intraoperative blood loss [(90.2±5.3) mL] was significantly less in the hook plate group than those in the reconstruction group [(95.6±20.8) min, (12.4±0.9) cm and (74.2±3.5) mL] ( P<0.05). There was no significant difference in hospital stay between the 2 groups ( P>0.05). At 3, 6 and 12 months after operation, the VAS scores (1.8±0.5, 1.2±0.3 and 1.1±0.2) and Constant-Murley scores (85.2±4.6, 91.1±2.6 and 92.1±2.2) in the reconstruction group were significantly better than those in the hook plate group (3.2±1.0, 1.6±0.3 and 1.5±0.3; 73.6±2.9, 85.9±4.6 and 87.0±3.1) ( P<0.05). At the last follow-up, the postoperative coracoclavicular separation ratio (elevation) in the hook plate group (0.20±0.16) was significantly greater than that in the reconstruction group (0.10±0.05) ( P<0.05). Conclusion:In the treatment of Neer ⅡB distal clavicular fractures, coracoclavicular ligament anatomical reconstruction may lead to better fixation and fewer postoperative complications than a clavicular hook plate, demonstrating fine clinical efficacy.

Objective:To construct human immortalized keratinocytes stably expressing human papillomavirus type 16 (HPV16) E6/E7 gene, and provide a cell model for studying mechanisms underlying HPV16 E6/E7-induced cell immortalization and malignant transformation.Methods:Primary human foreskin keratinocytes (HFKs) were isolated by sequential two-step enzymatic digestion. Cultured HFKs were stably transfected with a HPV16 E6/E7 gene-overexpressing lentiviral vector LV5-HPV16 E6/E7, and consecutively cultured for more than 30 passages. Then, immortalized keratinocytes were screened out and divided into 3 groups: (1) blank control group: second-passage primary HFKs; (2) experimental group: HFKs transfected with LV5-HPV16 E6/E7 at different passages, and the second-passage primary HFKs transfected with LV5-HPV16 E6/E7 were referred to as A0 cells, thereafter, the transfected HFKs were named according to their passage number, such as A1, A2, ... A30; (3) positive control group: the HPV16-positive cervical cancer cell line SiHa. Real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the mRNA expression of HPV16 E6/E7 and protein expression of HPV16 E6/E7 and CK14, respectively, in the blank control group, experimental group and positive control group. Cell counting kit-8 (CCK8) assay and Transwell insert invasion assay were conducted to assess the cellular proliferative and invasive activity. In vivo tumor formation experiment in nude mice was conducted to investigate the tumorigenicity of A30 cells in the experimental group and SiHa cells in the positive control group. Results:Primary HFKs were successfully isolated. After the primary HFKs were transfected with the recombinant plasmid LV5-HPV16 E6/E7, the blank control group showed no fluorescence in the cells, but showed senescent phenotypes after serial passages, while in the experimental group, the volume and morphology of A30 cells were similar to those of the primary HFKs with the proportion of fluorescence-positive cells being 100%. Compared with the blank control group, the experimental group showed significantly increased mRNA expression levels of HPV16 E6 and E7 in A1, A10, A20 and A30 cells (HPV16 E6: t = 7.12, 8.07, 6.53, 5.66, P < 0.001, < 0.001, = 0.001, = 0.005, respectively; HPV16 E7: t = 3.20, 4.29, 3.75, 4.22; P = 0.024, 0.008, 0.013, 0.014, respectively) . The protein expression of HPV16 E6/E7 was absent in the blank control group, but was observed in A30 and SiHa cells. CCK8 assay showed that the proliferative activity of A10, A20 and A30 cells was significantly higher than that of the blank control group ( t = 6.49, 7.55, 9.43, P = 0.003, 0.002, 0.001, respectively) , while there was no significant difference in the proliferative activity between A1 cells and the blank control group ( t = 2.40, P = 0.074) . Transwell insert invasion assay showed that A30 cells could not cross the basement membrane, but SiHa cells could pass through the basement membrane and were stained blue. Two months after the inoculation with A30 cells in the nude mice, no visible tumors were found, which was confirmed by a histological study. Subcutaneous tumors were formed in the nude mice after the inoculation with SiHa cells. Conclusion:Human immortalized keratinocytes were successfully established by lentivirus-mediated transfection with HPV16 E6/E7 gene, and can serve as an ideal cell model for HPV-related research.