Objective To investigate and optimize the condition of purification and crystallization of oncogenic protein MDM2.Methods MDM2 was expressed in E.coli expression system, and purified by Ni-NTA chelating affinity chromatography and molecular sieve chromatography.The secondary structure of purified protein was analyzed by circular dichroism spectroscopy (CD).Then the crystallization condition of MDM2 was screened and optimized by sitting-drop vapor-diffusion method.Results High purity of MDM2 was obtained by Ni-NTA chelating affinity chromatography and molecular sieve chromatography purification.CD analysis indicated the secondary structure of MDM2 was ordered.Protein-crystallisation experiments illustrated that MDM2 was prone to crystallization under lower pH.Conclusion The optimum pH of MDM2 protein crystallization is 5.5, the optimum protein concentration is 10 mg/mL.

Objective To observe the mechanisms of autophagic eukaryotic cells in Acinetobacter microvilli removal and protein histological study on apoptosis induced by macrophages .Methods A model of Acinetobacter baumannii infection was established in 24 female OCR mice .The mice were randomly divided into control group (n= 12) and observation group (n= 12) .The control group was injected with normal saline ,and the observation group was injected with autophagy eukaryotic cells ,the histopathological changes of Acinetobacter and the induction of macrophage apoptosis were observed .Results There was no significant difference in the bacterial counts between the two groups of mice immediately after implantation (P>0 .05) ,the bacterial counts in the 24 and 48 h in the observation group was significantly lower than that in the control group (P<0 .05) .The lung tissue of mice in the ob-servation group injected after autophagy was normal ,the alveolar cavity was open ,no abnormal substances were found ,the alveolar wall was not obviously thickened ,and no inflammatory cell infiltration was found in the wall .The mice in the control group were in-jected with normal saline and lacked the ability to remove Acinetobacter ,resulting in a large number of inflammatory cell infiltra-tion ,vasodilatation ,and congestion in some mice .Conclusion Autophagic eukaryotic cells injected with Acinetobacter baumannii can increase the clearance rate ,induce apoptosis of macrophages and improve the quality of Acinetobacter baumannii .

Objective:To clarify the characteristics of renal cortical microcirculation and its relationship with the expression of plasma endothelial microparticle (EMP) in septic rats, and to evaluate the effect of Xuebijing injection as an adjuvant therapy of antibiotics on septic AKI.Methods:The 8-10 weeks old specific pathogen free (SPF) male Sprague-Dawley (SD) rats were divided into sham operation group (Sham group), positive drug control group and Xuebijing group by the random number table method, with 10 rats in each group. The cecal ligation and puncture (CLP) with large ligation (ligated 75% of the cecum) was used to prepare a rat high-grade sepsis model; in the Sham group, the cecum was stretched without ligation or puncture. Due to the high mortality of CLP with large ligation, Xuebijing injection (4 mL/kg, 12 hours per time) and imipenem/cilastatin injection (90 mg/kg, 6 hours per time) were administered to the rats in the Xuebijing group via the tail vein immediately after the model was produced. Normal saline and imipenem/cilastatin were administered to the rats by the same methods in the positive drug control group. The rats in the Sham group were treated with the same volume of normal saline as any of the other two groups at the same frequency. At 48 hours after model reproduction, the mean arterial pressure (MAP) and blood lactic acid (Lac) of the rats were measured. The renal cortical microcirculation was monitored by using side stream dark-field imaging. Renal hypoxia signals were assessed by pimonidazole chloride immunohistochemistry. Plasma EMP levels were determined by using flow cytometry, and then the correlation between EMP and microcirculation parameters of renal cortex was analyzed. At the same time, the serum creatinine (SCr) was measured, and the renal injury score (Paller score) was used to evaluate the severity of renal tissue pathological damage.Results:Compared with the Sham group, perfused vessel density (PVD), microvascular flow index (MFI) and MAP in the positive drug control group and the Xuebijing group decreased significantly, the positive expression of hypoxia probe (pimonidazole) increased, Lac, EMP, Paller score and SCr increased significantly. However, compared with the positive drug control group, the renal cortical microcirculation in the Xuebijing group was improved significantly, PVD and MFI were increased significantly [PVD (mm/mm 2): 16.20±1.20 vs. 9.77±1.12, MFI: 2.46±0.05 vs. 1.85±0.15, both P < 0.05], Lac was reduced significantly (mmol/L: 4.81±1.23 vs. 6.08±1.09, P < 0.05), MAP increased slightly [mmHg (1 mmHg = 0.133 kPa): 84.00±2.00 vs. 80.00±2.00, P > 0.05], suggested that Xuebijing injection improved renal microcirculation perfusion in septic rats, and this effect did not depend on the change of MAP. The positive expression of pemonidazole in renal cortex of the Xuebijing group was significantly lower than that of the positive drug control group [(35.89±1.13)% vs. (44.93±1.37) %, P < 0.05], suggested that Xuebijing injection alleviated renal hypoxia. The plasma EMP levels of rats in the Xuebijing group were significantly lower than those in the positive drug control group (×10 6/L: 3.49±0.17 vs. 5.78±0.22, P < 0.05), and the EMP levels were significantly negatively correlated with PVD and MFI ( r values were -0.94 and -0.95, respectively, both P < 0.05), indicated that the increase of plasma EMP was highly correlated with renal microcirculation disorder, and Xuebijing injection inhibited the increase of plasma EMP levels. The Paller score in the Xuebijing group was significantly lower than that in the positive drug control group (46.90±3.84 vs. 62.70±3.05, P < 0.05), and the level of SCr was also significantly lower than that in the positive drug control group (μmol/L: 121.1±12.4 vs. 192.7±23.9, P < 0.05), which suggested that Xuebijing injection relieved kidney injury and improved renal function in septic rats. Conclusion:As an adjuvant therapy of antibiotics, Xuebijing injection could inhibit the expression of plasma EMP in rats with sepsis, improve renal cortex microcirculation, and reduce kidney injury.

Objective: To investigate the mid-term outcomes of the dual mobility total hip prosthesis in primary total hip arthroplasty. Methods: A total of 101 patients who underwent primary total hip arthroplasty with dual mobility total hip prosthesis from May 2010 to March 2013 were enrolled in the present study with complete follow-up information. There were 56 females and 45 males with the mean age of 66.36 years (rang from 58 to 77 years). There were 35 patients with femoral neck fracture, 33 patients with femoral head necrosis, 10 patients with hip osteoarthritis, 18 patients with secondary osteoarthritis to hip dysplasia and 5 patients with ankylosing spondylitis. The dual mobility total hip prosthesis was used for all 101 patients by the posterior-lateral approach of hip joint. Harris hip score was used to evaluate the clinical effects. Radiographic analysis was also performed to evaluate the biological fixation effects of the prosthesis, dislocation of the prosthesis, osteolysis and migration of the acetabular cup. Results: The mean operation duration was 80.68±6.59 min (range from 70 to 90 min). The average blood loss during the operation was 180.67±18.76 ml (range from 150 to 200 ml). All incisions were healed at the first stage. All patients were followed up with an average of 65±3 months (range from 62 to 75 months). Harris hip score improved from 56.70±16.71 before surgery to 94.26±1.91 at the last follow-up. The differences among different follow-up times were of statistical significance. The Hip flexion and extension, adduction and abduction, and internal and external rotation improved from 86.67°±16.70°, 34.06°±7.05°, 34.53°±7.45° preoperatively to 141.73°±6.56°, 57.06°±3.83°, 75.18°±4.00° at the last follow-up, respectively. At 3 months after the operation, the X-ray showed satisfied bone integration. There were no acetabular and femoral shaft fractures, sciatic nerve, femoral artery and vein injuries during the operation, and no joint dislocation, prosthesis loosening, infection and deep venous thrombosis of lower limbs during the postoperative and follow-up duration. Conclusion The dual mobility total hip prosthesis has the advantages in good initial and middle stability, rapid bone growth, low dislocation rate and repid recovery of postoperative motion range. It is suitable for patients over 65 years old or younger patients with postoperatively high dislocation.

Objective To analysis the change of hip joint in healthy infants by ultrasound,and establish the normal reference value of the developmental dysplasia of the hip(DDH). Methods A total of 8 000 healthy infants from 0 to 24 weeks were collected from the Multi-center study of 10 children′s medical centers. Among them,3 855 infants(2 065 females and 1 790 males) with complete data and follow-up were included in this study. All subjects were divided into 6 groups ( ＜4,4～7,8～11,12～15,16～19 and≥20 weeks group). α angle,femoral head length and width,femoral head coverage ratio by acetabulum ( FHC) were measured in the coronal view on the neutral position;distance from pubis to femoral head ( P-H) and distance from ischium to femoral head ( I-H ) were measured in the transverse view on neutral position;distance from femoral head topubis ( H-P) was measured in the posterolateraltransverse view on the flexion position. The results of each group changes with age were analysised. Results ① The α angle of healthy infants from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05),but there was no significant difference between 16～19 and ≥20 weeks group( P ＞0.05). ②The femoral head length and width of all age groups were increased with age,the difference among all the groups was statistically significant( all P ＜0.05). ③ FHC from 0 to 20 weeks were increased with age,the difference among the groups were statistically significant( P ＜0.05) except between 16～19 and ≥20 weeks group( P ＞0.05). ④ The P-H and I-H in all age groups showed no statistically significant ( all P＞0.05). ⑤The H-P of all age groups were increased with age,the difference between the groups were statistically significant(all P ＜0.05).Conclusions The development of hip joints have the certain regular developmental pattern in healthy infants less than 5 months of birth and are relatively constant after birth more than 5 months. The ultrasound normal reference value of the hip joints can be used for the early diagnosis of the DDH.

Objective To explore the influence of the iodine disinfection on nasal bacterial colonization through the transsphenoidal approach. Methods Totally 133 pituitary adenoma patients who underwent transsphenoidal surgery in our department from January to August 2017 were enrolled in this study. Before disinfection,pharyngeal swabs of inferior turbinate root secretions were taken for bacterial culture. After iodine disinfection,pharyngeal swabs were taken again at the same site. Changes in the nasal bacterial spectrum before and after disinfection were compared. Patients were followed up for three months after the surgery,during which any intracranial infection/bacteraemia was recorded,and its correlation with nasal bacteria colonization was analyzed. Results Nasal bacterial colonization was detected in 45 (33.8%) of 133 patients before iodine disinfection and in only 6 cases (4.5%) after iodine disinfection (χ=34.5,P=0.000). Thus,iodine disinfection eliminated 86.7%(39/45) of the colonized bacteria. The most common nasal bacterium was Staphylococcus aureus (24.4%,11/45),followed by Klebsiella pneumoniae (24.4%,11/45),and Staphylococcus epidermidis (13.3%,6/45). One patient had high fever and chills 2 days after surgery,but blood culture and cerebrospinal fluid culture showed negative Results . After the administration of third-generation cephalosporins,the symptoms disappeared after two days. Conclusion sThere are colonized bacteria in nasal cavity. Iodine disinfection of nasal cavity can effectively clear most of the nasal bacteria. The possibility of intracranial infection/bacteremia after transsphenoidal approach is low.