Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

Objective:To investigate the interventional effects of the Fuzheng Huayu (FZHY) formula and its partial mechanistic role on cholestatic liver fibrosis in mice.Methods:Mdr2 gene knockout (Mdr2－/ －) mice were randomly divided into a model group, FZHY group, and Obeticholic acid group. Wild-type C57BL/6J mice of the same age served as the control group. Mdr2－/ －mice were given the corresponding drugs starting from the first day of 9 weeks of age by oral gavage in each group. The control and model groups were administered 0.3% sodium carboxymethylcellulose by oral gavage and were sacrificed at 12 weeks of age for specimen collection. High-speed biochemistry analyzer was used to detect serum alkaline phosphatase and alanine aminotransferase activity in mice. Hematoxylin-eosin staining and Sirius red staining were used to observe pathological changes in liver tissues. Hydroxyproline content was measured to assess collagen in liver tissues. Immunohistochemical staining, Western blotting, and real-time fluorescence quantitative PCR were used to detect the expression of fibrosis markers Col-I and alpha-smooth muscle actin in liver tissues. The expressional condition of cholangiocyte response markers Epcam, CK7, CK19, as well as Pcna, Mki67, and Ccnd1, inflammatory related factors Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4, phosphorylated peroxisome proliferator-activated receptor alpha (PPARα) and nuclear factor kappa-B (NF-κB) were determined. Comparative analysis among multiple groups was performed using one-way ANOVA. The LSD method was used for comparisons between groups. Two-tailed statistical tests were used.Results:Compared with wild-type mice, Mdr2 －/ － mice had a significant increase in serum alanine aminotransferase and alkaline phosphatase activity ( P<0.001). The percentage of Sirius red-positive staining areas and hydroxyproline content in liver tissues was significantly increased ( P<0.01). The expression of Col-I, α-smooth muscle actin, Epcam, CK7, CK19, Pcna, Mki67, and Ccnd1, and the expression of Ccl2, Ccl5, Tnf-α, Il10, and Cxcl4 were significantly increased ( P<0.01); however, both FZHY and Obeticholic acid significantly reversed the increases in these indicators ( P<0.05; P<0.01). Further results showed that compared to wild-type mice, the expression of PPARα was significantly reduced in liver tissues of Mdr2 －/ － mice, while NF-κB was significantly enhanced ( P<0.01). In contrast, compared to Mdr2-/- mice, the expression of PPARα in the liver tissues of FZHY group mice was significantly increased ( P<0.05), while NF-κB was significantly inhibited ( P<0.05). Conclusion:FZHY can significantly improve liver fibrosis, cholangiocyte response, and inflammation in Mdr2 －/ － mice with spontaneously occurring cholestatic liver fibrosis, and its mechanistic role is related to the regulation of the PPARα/NF-κB pathway.

【Objective】 To study the effect of macrophage mediator 1 (MED1) deficiency on atherosclerosis in female mice. 【Methods】 ApoE knockout (ApoE^-/-), LDLR knockout (LDLR^-/-), MED1^fl/fl, and macrophage MED1 knockout (MED1^△Mac) mice were recruited in the study. Two types of mouse model were constructed:ApoE and macrophage MED1 double knockout (MED1^△Mac/ApoE^-/-) mice and their littermate controls (MED1^fl/fl/ApoE^-/-). ② LDLR knockout (LDLR^-/-) mice receiving bone marrow from MED1^△Mac (MED1^△Mac→LDLR^-/-) or MED1^fl/fl (MED1^fl/fl→LDLR^-/-) mice. Female mice from these two models were fed a Western diet (21% fat and 0.15% cholesterol) for 12 weeks to promote the development of atherosclerosis. Body weight, total cholesterol (TC), and total triglyceride (TG) content in plasma were measured dynamically. After Western diet feeding for 12 weeks, aortic tree and aortic root were collected and hematoxylin-eosin (H&E) and oil red O staining were performed. 【Results】 Plasma TC and TG did not significantly differ between MED1^fl/fl/ApoE^-/- control group and MED1^△Mac/ApoE^-/-experimental group. However, the plaque area in aortic tree and aortic root was significantly increased in MED1^△Mac/ApoE^-/-mice. Moreover, compared with that in MED1^fl/fl→LDLR^-/- control group, the plaque area of aortic tree and aortic root had an increasing trend in MED1^△Mac→LDLR^-/- mice group. 【Conclusion】 MED1 deficiency in macrophages promotes the development of atherosclerosis in female ApoE or LDLR knockout mice.

Objective:To explore the morphological characteristics, treatment strategies and clinical results of complex hyperextension tibial plateau fractures.Methods:From October 2017 to January 2019, data of 27 patients with complex hyperextension tibial plateau fractures were retrospectively analyzed. There were 19 males and 8 females with an average age of 43.4 years (range, 23-68 years). According to Schatzker classification of tibial plateau fractures: there are 8 cases of type IV, 5 of type V, and 14 of type VI; according to the three-column theory classification: there are 8 cases of two-column fracture and 19 cases of three-column fracture. Bicondylar fractures were treated with medial Tomofix locking plate and anterolateral L-shaped locking plate through medial and anterolateral approach; tibialmedial condylar fractures was treated with T-shaped plate and posteromedial locking plate through extended medial approach. Patients with anterior tibial fractures were treated with horizontal strip plate through modified anterior median approach. Combined soft tissue or bone injury was repaired. The fracture healing and reduction were evaluated by X-ray and CT scan. The reduction of tibial plateau fracture was evaluated by Rasmussen radiology standard, and the knee joint function was evaluated 12 months after the operation by the score of American hospital for special surgery (HSS).Results:All the 27 surgeries were performedsuccessfully. The operation time was 130-350 minutes, with an average time of 165 minutes. Twenty-seven cases were followed up for 12-24 months, with an average period of 15.8 months. All fractures were healed. The average clinical healing time was 13.5 weeks (range, 10-18 weeks). Twelve months after operation, Rasmussen's radiology score was 13-18, with an average of 16.7 points, among them there were 19 excellent and 8 good. Twelve months after the operation, the score of HSS knee joint was 82-98, with an average score of 93.2 points, and there were 22 cases excellent, 4 cases good and 1 case fair. The excellent and good rate was 96.2% (26/27).Conclusion:Complex hyperextension tibial plateau fractures often combined with tibial bicondylar, medial tibial condyle or anterior tibial fractures. According to the morphological characteristics of complex hyperextension tibial plateau fractures, using appropriate surgical approach and internal fixation, repairing ligament soft tissue structure and reconstructing knee joint stability can achieve satisfactory results.

Objective:To discuss how to make the surgical strategy for tibial tubercle fracture associated with bicondylar tibial plateau fracture.Methods:Data of thirty-five patients of tibial tubercle fractures associated with bicondylar tibial plateau fractures who were treated from October 2014 to May 2018 were retrospectively analyzed. There were 26 males and 9 females with an average age of 37.6 years (range, 21-68 years). According to Schatzker classification in tibial plateau fracture, 16 cases were type V and 19 cases were type VI. According to the integrity of tibial tubercle fracture and cortical bone of the proximal tibia in bicondylar tibial plateau fracture, they were divided into four types: type A, tibial tubercle fracture fragment and cortical bone of the proximal tibia are both complete; type B, tibial tubercle fracture fragment is complete but cortical bone of the proximal tibia is comminuted; type C, tibial tubercle fracture fragment is comminuted but cortical bone of the proximal tibia is complete; type D, both of them are comminuted. The surgical approaches and fixation methods of all the tibial tubercle fractures were according to the four different types. There were 22 cases with type A and B that were treated via an anterolateral and a medial incision, 13 cases with type C and D were treated via an anterior midline and a medial incision. There were 4 cases belonging to type A fixed with lag screws singly, 18 cases with type B fixed with 1/4 tubular plates, 7 cases with type C and 6 cases with type D fixed by 1/4 tubular plates combined with lag screws.Results:Thirty-five patients were followed up for 16.8 months (range, 12-24 months). All fractures healed with an average time of 4.7 months (range, 3-6 months). Loss of reduction didn’t occur in 34 cases except one. According to Rasmussen radiographic evaluation, the average score was 14.1 (range, 10-18) and clinical outcomes were rated with excellent in 11 cases, good in 19, fair in 5. The excellent and good rate was 85.7% (30/35) . The mean Hospital for Special Surgery (HSS) scores of all cases were 86.8 (range, 64-98) and the functional scores were excellent in 22 cases, good in 10 cases and fair in 3 cases with the excellent and good rate of 91.4% (32/35) . Surgical complications included fat liquefaction in 2 cases, superficial wound infection in 1, loosening of implant in 1and traumatic arthritis in 1.Conclusion:This kind of tibial tubercle fracture associated with bicondylar tibial plateau fracture is rare and special. Therefore, the preoperative plan should be made by considering the morphological features of the tibial tubercle fragments and the cortical bone of the proximal tibia. The middle longitude approach is the best way to expose tibial tubercle fragments which should be fixed with 1/4 tubularplate and/or lag screws.

Objective: To explore the clinical features and treatment strategies of tibial plateau fractures sustained with hyperextension varus. Methods: Data of 11 patients of tibial plateau fractures with hyperextension varus treated from January 2008 to November 2017 were retrospectively analyzed. There were 7 males and 4 females with an average age of 41.2 years old (range, 25-67 years). Injuries were caused by falling down in 7 cases, traffic accident in 3 cases, and falling from height in 1 case, respectively. On the basis of Luo's three columns classification in tibial plateau, there were 9 cases of medial column fracture and 2 cases of medial combined with posterior column fracture. Six cases were concomitant with fibular head fracture and 2 cases with the injury of common peroneal nerve. Preoperative magnetic resonance imaging showed that there were anterior cruciate ligament injury in 3 cases, posterior cruciate ligament injury in 4 cases, medial meniscus injury in 5 cases, lateral meniscus injury in 3 cases, medial collateral ligament injury in 6 cases, iliotibial band injury in 2 cases and posterolateral complex injury of the knee joint in 9 cases, respectively. All tibial plateau fractures were treated firstly by open reduction and internal fixation via medial approach of the knee. The medial meniscuses and collateral ligaments were explored, in which of them there were 2 medial meniscuses with the marginal tear been sutured simultaneously. Then the knee joints which were still unstable after the examination of stable tests in 6 cases with posterolateral complex injuries were repaired surgically via lateral approach. Fibular head fractures were fixed with anchor nails or cannulated screws in 6 cases. The ruptured posterior cruciate ligaments in 2 cases were reconstructed with autologous tendon transplantation under endoscopy. Results: All the patients were followed up for an average period of 16.2 months (range, 12-22 months). All fractures were healed in 10-20 weeks with an average time of 16.5 weeks. The range of extension of the affected knee joint in all patients was 0° and the average flexion was 135° (range, 120°-145°) one year after surgery. The average flexion of affected knee in 4 cases which were only treated with the tibial plateau fracture without the mild ligament injuries was 137° (range, 132°-145°) and the average flexion of affected knee in 7 cases who were treated with tibial plateau fracture and severe posterolateral complex included posterior cruciate ligaments completely broken with reconstruction was 132° (range, 120°-140°). According to Rasmussen radiographic evaluation, the average score of all patients was 16.3 (range, 14 to 18) and clinical outcomes were rated with excellent in 10 cases and good in 1. The excellent and good rate was 100% (11/11). The mean of the hospital for special surgery (HSS) score was 86.7 (range, 79-96) and the functional scores were excellent in 9 cases, good in 2 cases thus the excellent and good rate was 100% (11/11). Both varus stress test 30° and dial test were positive in one case considered for the ligament laxity postoperatively who didn’t accept further treatment and the stabilization tests were negative in the other 10 cases. There were no intraoperative complications in all patients such as neurovascular injury. No incision infection, failure of the implants and fracture nonunion occurred postoperatively. Traumatic arthritis of the affected knee occurred one year after surgery in 1 case who had no obvious pain after treated with oral medicine. Conclusion The hyperextension varus injuries of the knee are rare clinically. The posterolateral complex should be evaluated thoroughly for this injury pattern. If it's necessary, the posterolateral structures must be repaired surgically after the tibial plateau fractures are fixed.

Objective To explore the guiding significance of posterior tibial plateau partition for the selection of surgical approach in treatment for posterior column fracture.Methods From June 2008 to May 2015,46 patients with posterior column fractures of tibial plateaus treated were retrospectively analyzed.There were 31 males and 15 females with an average age of 35.1±12.8 years old (range,19-62 years).Nineteen patients were left side and twenty-seven patients were right side.Injury was caused by traffic accident in 27 cases,falling from bicycle in 12 cases and falling from height in 7 cases.On the basis of the posterior condyle with anatomical structure in tibia1 plateau,the posterior column was divided into four parts.All fractures were treated via the optimal approach based on the location of them.The posteromedial approach was used with fractures in zone 1 and 2,posterolateral approach used in zone 3,lateral approach via the fibular head osteotomy was used in zone 4 and combined approaches were used in multiple zones.All the fractures involved the posterior column were treated by anatomical reduction and fixation with plates and screws under direct vision.Results All the patients were followed up with an average of 15.5±3.7 months (range from 12 to 24 months).The healing time of all patients was 11-18 weeks,with an average time of 14.6±2.3 weeks.According to Rasmussen radiographic evaluation,the average score was 15.1 (range from 11 to 18) and clinical outcomes were rated with "excellent" in 17 cases,"good" in 24 cases,"fair" in 5 cases.The excellent and good rate was 89.1％ (41/46).The mean HSS (the Hospital for Special Surgery) score of all patients at 12 months operatively were 86.7±8.6 (range from 67 to 98) and the functional scores were excellent in 25 cases,good in 17 cases and fair in 4 cases with the excellent and good rate was 91.3％ (42/46).The average range of motion in affected knee was 118°±13.7° (range from 0° to 135°) in 17 cases via posterolateral approach,123°±15.6° (range from 0° to 135°) in 18 cases via posteromedial approach,115°±16.7° (range from 0° to 130°) in 18 cases via combined posteromedial and posterolateral approaches and 124°±7.4° (range from 0° to 130°) in 4 cases via the fibular head osteotomy lateral approach.Complications included fat liquefaction in 1 case,anterior tibial artery spasm in 1 case and traumatic arthritis in 1 case.Conclusion The partition of posterior tibial plateau can be used to guide the surgical approach to the posterior column simply and accurately.For the fractures of isolated posterior column and posterior column mainly involved,the partition has a certain guiding significance.

Bovine adenovirus type 3 (BAdV3) is being used in the development of potential vehicles for gene therapy and vectored vaccine. To that end, a more comprehensive description of BAdV3 biology is essential. In this study, we focused on the role of pIX in BAdV3 virion rescue after full-length BAdV3 genome transfection. Initially, pIX deletion or initiation codon mutation abolished the production of progeny virions, which suggested that pIX was essential for the rescue of BAdV3 containing a full-length genome. Moreover, through transfection of a panel of pIX mutant BAdV3 genomes, we observed that the conserved N-terminus and the putative leucine zipper element (PLZP) were essential for virion rescue, whereas the C-terminus following the coiled-coil domain was non-essential. In addition, swap of the PLZP element and its following region of BAdV3 pIX to corresponding domains of human adenovirus type 5 (HAdV5) did not affect virion production, whereas swap of the entire pIX abolished production of progeny virions. We suggest that failure of the full-length BAdV3 pIX swap might be due to species specificity of its N-terminus region before the PLZP element.
Adenoviridae* ; Adenoviruses, Human ; Biology ; Codon, Initiator ; Genetic Therapy ; Genome ; Genome, Viral* ; Leucine Zippers ; Species Specificity ; Transfection* ; Virion*

Objective:To investigate the clinical application value of ultrasound in the diagnosis and differential diagnosis of Takayasu arteritis (TA ). Methods: A total of 39 suspected TA patients were examined by ultrasonography,erythrocyte sedimentation rate (ESR),C reactive protein (CRP)and CT angiography (CTA), and the ultrasound examination results were compared and analyzed with the final chinical diagnosis results. Results:Of the 39 cases of suspected TA patients which had been examined by ultrasound,33 cases were diagnosed as TA,2 cases were diagnosed as atherosclerosis (AS),2 cases were diagnosed as fibromuscular dysplasia (FMD) and 2 cases were diagnosed as thromboangiitis obliterans (TAO).Compared with final chinical diagnosis results, 1 FMD patient was misdiagnosed as TA,and the rest of the diagnostic results were consistent with the final chinical diagnosis results.Conclusion:Ultrasound examination can provide objective and accurate basis for the diagnosis and differential diagnosis of TA,and it is an effective method of imaging examination.

OBJECTIVETo investigate the effects of PPARγ overexpression on steatosis in mouse primary hepatocytes.

METHODSPrimary hepatocytes isolated from C57BL/6J mice were infected with either Ad/LacZ or Ad/PPARγ for 48 h. Steatosis of the primary hepatocytes was checked by Oil Red O staining. The mRNA and protein expression of adipocyte-specific genes PPARγ, aP2 and CideA were analyzed by using RT Real-time PCR and Western Blot.

RESULTSPrimary hepatocytes were small and even. Hepatocyte nuclei were round with dispersed chromatin and prominent nucleoli. Accumulated lipid droplets were observed in Ad/PPARγ-infected hepatocytes, but in Ad/LacZ-infected hepatocytes. Moreover, compared with Ad/LacZ-infected hepatocytes, the mRNA expression of PPARγ, aP2, FGF21 and CideA in Ad/PPARγ-infected hepatocytes were significantly induced, the protein expression of PPARγ and its target aP2 strongly increased.

CONCLUSIONover expression of PPARγ induces adipogenic steatosis in mouse primary hepatocytes.

Adipocytes ; metabolism ; Adipogenesis ; Animals ; Cells, Cultured ; Fatty Liver ; metabolism ; pathology ; Genetic Vectors ; Hepatocytes ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; PPAR gamma ; metabolism ; Transfection