To capture a state of the enzyme in complex with an intact substrate, we developed and adopted a novel freezing method in crystal preparation procedure. Neither the elimination of the catalytically indispensable ligands, nor mutation or modification of the active site is required. At -20 degrees C, we soaked the crystal of 6-phosphate-beta-glucosidase (Bg1A) in the liquor containing p-nitrophenyl-beta-D-glucopyranoside-6-phosphate (pNPbetaG6P). The diffraction data at 2 A resolution was collected and an intact and unambiguous electron density map of pNPbetaG6P was obtained. These results provide an effective method for the research of cryoenzymology and the intermediate state of enzyme-substrate complex in the future.
Cold Temperature ; Crystallization ; Glucosidases ; chemistry ; metabolism ; Substrate Specificity ; X-Ray Diffraction

Osteoporotic spinal fractures is a serious orthopedic chronic disease in the healthy elder people .Because of the complexity of its pathogenesis , there has no standard treatment scheme .In order to solve this problem , we review the mechanism of the disease etiology, imaging, treatment, prevention of osteoporotic spinal fractures to provide references for the further prevention and con -trol of this disease .

Objective To observe whether peripheral blood mononuclear cells (PBMCs) from patients with active systemic lupus erythematosus (SLE) can induce the adhesion of T lymphocytes to endothelial cells. Methods Sera and PBMCs were obtained from patients with active SLE and normal human controls. PBMCs were cultivated and culture supematants were harvested. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro with or without the presence of the sera or culture super-natants of PBMCs. Some cells were pretreated with the antibody to IL-17 before the treatment with the sera or supematants. After another 48-hour culture, RT-PCR and real-time PCR were used to detect the mRNA expressions of adhesion molecules, including intercellular adhesion moleeule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-cadherin in HUVECs, wound healing assay to estimate the motility of HUVECs. Additionally, T lymphocytes were added to HUVECs 48 hours after stimulation with the sera or supematants, the adhering of T lymphocytes to HUVECs was observed by microscopy. Results After stimulation with supematants of PBMCs from patients with active SLE, the mRNA expressions of ICAM-1, VCAM-1 and E-cadherin were significantly increased in HUVEC, while the increase could be inhibited by the antibody to IL-17. The elevation of adhesion molecule expression subsequently promoted the motility of HUVEC, mediated the adhesion of T lymphocytes to HUVEC, and the antibody to IL-17 could suppress the adhesion of T lymphocytes and motility of HUVEC. Conclusion The culture supematants of PBMCs from patients with active SLE can induce the expression of vascular cell adherin molecules and promote the adherin of T lymphocytes, which may in turn mediate the development of lupus vasculitis.

OBJECTIVE To find out the situation of the usage of antimicrobials in type Ⅰand type Ⅱ operations during perioperative period and to evaluate the rationality of use. METHODS To check 472 examples of medical record data of discharged patients with type Ⅰand type Ⅱ operations in each department during from Apr to Jun and from Aug to Oct 2006,and make statistics and analysis according to application rate of antibacterial drugs, examining rate of the samples, drug usage by experiences and postoperative infection rate. RESULTS The application rate of antibacterial drugs during perioperative period in our hospital was high;the examining rate of correlated samples was low;the choice of antibacterial drugs mainly depended on experience;the sensitivity of the chosen antibacterial drugs was not strong;the duration of prophylactic drug usage was too long;and the postoperative infection rate was high. CONCLUSIONS It is very important to strengthen standardized administration of prophylactic antibiotic usage during perioperative period in our hospital.

Objective To observe the diagnostic value of telomerase,VEGF,CD44_ V6 ,MMP_2,MMP_9and their combined examination in distinguishing malignant pleural effusion from benign one.Methods Sixty patients with pleural effusions were observed,thirty of them sufferring from malignant pleural effusion and thirty of them sufferring from benign one.The expression of telomerase,CD44_ V6 ,VEGF,MMP_2,MMP_9were tested in the pleural effusion by RT-PCR and ELISA technique.The sensitivity and the specificity of each examination method were calculated,in order to find out appropriate combination.Results As single-item detection,the diagnosetic value of VEGF was the best,the cutoff of VEGF was 181.1pg/ml,the sensitivity of VEGF examination was 90.0%,and the specificity was 86.7%.The sensitivity and the specificity of combined examination VEGF+CD44_ V6 and VEGF+MMP_2 were the best,the sensitivity of VEGF+CD44_ V6 was 83.3%,and the specificity was 90.0%.The sensitivity of VEGF+MMP_2 was 80.0% and the specificity was 93.3%.Conclusion The level of telomerase,VEGF,CD44_ V6 ,MMP_2,MMP_9in malignant pleural effusion is clearly higher than that in benign pleural one,the difference having significant statistic meaning.VEGF has higher sensitivity and specificity .Of combined examination,VEGF+CD44_ V6 and VEGF+MMP_2 have higher sensitivity and specificity.

Objective:To study the application of herbs of National TCM Master Li Jiren on treating atrophy with deficiency syndrome. Methods:By collecting 195 cases of atrophy-disease treated by Professor Li in Out-patient Medical Record System dated from January 2013 to December 2018, and FP-Growth computing paradigm was used to analyze frequency, FP-Growth enhanced computing paradigm was used to analyze the frequencyand association of symptoms and herbal pairs. Professor Li’s experience was accordingly summurized with clinic situation and Traditional Chinese Medicine theory. Results:There are 121 main symptoms among 195 selected cases, and amyotrophy (79 times), decreased muscle strength (69 times), difficulty in walking (52 times), weakness of upper extremities (43 times) and poor sleeping quality (33 times) rank the top 5 symptom. In terms of tongue manifestations, the color pink, pale, red and purple are the most; while in terms of pulses, weak, thread, deep and stringy were frequent. A total of 159 sorts of herbs were collected and Astragali Radix (140 times), Angelicae Sinensis Radix (103 times), Cistanches Herba (93 times), Psoraleae Fructus (87 times), Morindae Officinalis Radix (81 times) were the top 5. The herbs used are mainly with the function of tonifying deficiency, dispelling wind and dampness promoting blood circulation and removing blood stasisand. most of the herbs belong to liver, kidney and spleen meridians with warm character, sweet and bitter in taste. Association analysis between high-frequency herbs indicated that Astragali Radix is the core of most formulas, which conformed to Li’ s clinical experience. Three prescriptions were also obtained through cluster analysis. Conclusion:Professor Li treated atrophy diseases with deficiency syndrome by the unique idea of strengthening primordial qi and consolidating body resistance of Xin’an Medicine school. On the ground of using herbs to tonify, synergy of herbs, which were able todispel pathogenic wind, dredge channels and promote blood circulation for removing blood stasis, was achieved based on differentiation.

OBJECTIVE: To observe the effect of electro-acupuncture of Neiguan (PC 6) on mean systemic arterial blood pressure and plasmic concentrations of NO and TNFalpha in endotoxin shock rats. METHODS: The model of endotoxin shock was induced by lipopolysaccharide (1.5 mg/kg i.v.) and D-galactosamine (100 mg/kg i.p.). Aminoguanidine (100 mg/kg i.p.) and electro-acupuncture of bilateral Neiguan (PC 6) were administered. A catheter was inserted into the right subclavian artery to record the change of blood pressure and the blood was abstracted out and centrifuged to determine the NO and TNFalpha concentrations. RESULTS: Electro-acupuncture of Neiguan (PC 6) retrieved the blood pressure and reduced the plasmic NO and TNFalpha concentrations. CONCLUSION: Electro-acupuncture of Neiguan (PC 6) expresses an anti-endotoxin shock effect by repressing the plasmic NO and TNFalpha concentrations smoothly and retrieving the blood pressure stably.

Objective To investigate the effect of Plastrum Testudinis on neural stem cell after focal cerebral ischemia.Methods Rat models of focal cerebral ischemia were established by inserting nylon thread into the anterior cerebral artery.Immunohistochemical method was used to detect the expression of the neuroepithelial stem cell protein(Nestin),neurologic deficit and histological features were also observed.Results Plastrum Testudinis can decrease the neurologic dificit score and promote the expression of Nestin.Conclusion Plastrum Testudinis can alleviate the neurological symptoms and promote the proliferation of neural stem cell after focal cerebral ischemia.

Objective: To explore the effect of Plastrum Testudinis on the three subtypes of NOS (nNOS,iNOS,eNOS) expression in rats with focal cerebral ischemia. Methods:Rat model of focal cerebral ischemia was set up by inserting nylon thread into the internal carotid artery . The expressions of nNOS, iNOS and eNOS was detected by immunohistochemical method to observe the effects of Plastrum Testudinis. Results: Expressions of nNOS and iNOS in cerebral cortex and caudate-putamen of Plastrum Testudinis treatment group were significantly lower than those of model control group (P

Objective To investigate the efficacy and safety of the minimally invasive percutaneous surgery in the treatment of thoracolumbar fracture combined with posterior ligament injury.Methods The 94 cases of thoracolumbar fracture combined with posterior ligament injury who were admitted into our hospital from Septenber 2012 to January 2016 were selected and equally divided into the observation group and the control group(47 cases in each group) according to different methods.Patients of the observation group were treated with short segment of minimally invasive percutaneous surgery,while patients of the control group were underwent single segment of minimally invasive percutaneous surgery.Recorded the outcomes and prognosis of the two groups.Results There was no significant difference in the operation time and blood loss between the two groups(P ＞ 0.05).The postoperative volume of drainage and the length of stay in the observation group were significantly shorter than those in the control group(P ＜ 0.05).The JOA score of the observation group 1 month after operation was significantly better than that of the control group(P ＜ 0.05).The incidence of complications including pedicle screw breakage,pedicle screw loosening,incision infection,and pulmonary infection in the observation group was 4.3％,which was significantly less than 21.3％ in control group (P ＜ 0.05).The Cobb's angle of the obervation group and control group 1 month after operation were (8.55 ± 1.34) ° and (16.30 ± 2.33) ° respectively,which were significantly lower than (32.19 ± 1.98) ° and (32.08 ± 2.08) ° before operation (P ＜ 0.05).And the Cobb's angle 1 month after operation in the observation group were significantly lower than that of the control group(P ＜ 0.05).Conclusion The short segment of minimally invasive percutaneous surgery for the treatment of thoracolumbar fracture combined with posterior ligament injury can reduce the incidence of postoperative complications,promote the patient's rehabilitation,correct the kyphosis and promote the recovery of neural function,which was safer and more efficient.