According to the sequence of P450 cDNA of Eleutherococcus senticosus, specific primers were designed. Frokaryotic ex pression vector pET30a-P450 was constructed and the prokaryotic expression conditions were optimized. Results showed that the BL21 after being transformed with the recombinant expression vector accumulated the high amount of recombinant protein. SDS-PAGE analysis showed that the recombinant protein was about 53 kDa. The recombinant accumulated the highest amount of recombinant protein af ter IPTG (1 mmol x L(-1)) at 27-37 degrees C for 24 h. Consequently P450 gene of E. senticosus could be expressed successfully by prokaryotic expression vector pET30a-P450. Induction temperature, IPTG concentration, medium type and amount of induction time could all influence the expression of target protein, but the impact strength was different.
Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Eleutherococcus ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

Objective To evaluate the effects of different selection criteria on the prognosis of hepatocellular carcinoma(HCC)patients undergoing liver transplantation(LT)and to evaluate a new criterion.Methods A retrospective analysis was performed on 81 consecutive patients with HCC who underwent LT.The survival rates of the patients who met different criteria such as Milan.UCSF(University of California San Francisco UCSF).and Pittsburgh(Pitt)modified TNM criteria were calculated by KaplanMeier method,and the value of different criteria was evaluated.The Long-Rank test and COX proportional hazards regression model were performed to analyze the prognostic factors.the model of criteria was established according the most important prognostic factors.Using the Kaplan-Meier method,the suitable cut-offs of every variable ifl the model were found by comparing the survival and the number of the patients who met the cut-off,and considering the significant difference between the patients who met and exceed the cut-off at the same time.Resuits The 1,2,3-year accumulative survival rates of the 19 patients who met Milan criteria were 87.7%,87.7%,and 52.6%respectively:the 1,2,3-year disease free survival rates of them were 88.9%,72.7%,and 72.7%respectively.The 1,2.3-year accumulative survival rates of the 26 patients who met UCSF criteria were 87.2%,80.5%,and 55.2%respectively;the 1,2,3-year disease free survival rates of them were 84.1%,68.4%,and 68.4%respectively.With our new expanded criterion as of solitary tumor≤8 cm in diameter.or no more than 3 tumors,with the largest≤6 am,and a total tumor diameter≤10 cm.there was no significant difierence in 1,2,3-year sunrival rates and disease free survival rates(89.0%,81.8%,71.8%,and 81.9%,72.4%,72.4%.)as compared with Milan or UCSF criteria.but with this new criterion more patients(a=41)would be eligible for transplantation with a comparable long term survival.and the difference of the accumulative survival rates and disease free survival rates of the patients who met and exceed the new criteria was significant(P＜0.05).Conclusion The new indication is acceptable because the criteria does not adversely impact survival.

Objective To investigate the effect of microcireulation blood flow and altering immunity by Chuanxiongqin,ulinastain and thymosin α1 on sepsis patients.Methods 90 patients were randomly divided into 3 groups(n=30),namely ICU group,Chuanxiongqin group,ulinastain and thymosin α1 group.HLA-DR/CD14+and IL-6,TNF-α,Lac,DD were measured.Results (1)DD showed no significant difference at every time point between ICU group and ulinastain+thymosin α1 group(P＞0.05).DD decreased in Chuanxiongqin group,and was significantly different from the others on the third day.(2)Lac unchanged significantly at every time point in ICU group(P＞0.05).Lac in Chuanxiongqin group and ulinastain+thymeain α1 group tended to decrease,and was statistically different from ICU group on the second day.(3)IL-6 and TNF-α tended to increase at every time point in ICU group(P＜0.05).In ulinastain+thymosin α1 group,IL-6 and TNF-α returned to the level before treatment,HLA-DR/(D14+increased significantly,and was higher than Chuanxiongqin group and ICU group statistically.Conclusion Chuanxiongqin could ameliorate circulation;ulinastsin and thymosin α1 could depress IL-6,TNF-α.So ulinastain and thymosin α1 might protect the immunity of sepsis patients.

BACKGROUND: In the past ten years, because of the complications such as adjacent vertebral diseases after the discectomy and interbody fusion fixation, the artificial cervical disc replacement gradually replaces part of intervertebral disc and interbody fusion with internal fixation, and has become a new choice of a surgical treatment of spinal degenerative diseases resection; however, heterotopic ossification after the artificial intervertebral disc replacement as a postoperative problem has attracted the field attention. OBJECTIVE: To review the domestic and foreign research progress on heterotopic ossification after artificial disc replacement. METHODS: The first author retrieved PubMed database, Chinese Journal Full-text Database for related articles published from November 1973 to May 2017. The key words were "cervical vertebrae, artificial disc replacement, heterotopic ossification, research progress". A total of 216 articles were retrieved, and 45 articles met the inclusion criteria. RESULTS AND CONCLUSION: (1) Heterotopic ossification had a certain occurrence rate after artificial disc replacement. The susceptible factors and preventive measures were still controversial. (2) Future research regarding heterotopic ossification after artificial intervertebral disc replacement mainly focused on the development of biomechanics, molecular level and degenerative change. It is expected that in the near future, artificial disc that is more suitable for human spine biomechanics environment will appear; heterotopic ossification after artificial disc replacement will be more in-depth research.

Objective To evaluate the acute and delayed killing effect of high intensity focused ultrasound (HIFU) on Echinococcus granulosus(E. granulosus)protoscolices in vitro.Methods E. granulosus protoscolices were treated with different dosage of effective power(0,25,50,100,200,250 W)and time(5,10,20,30,40,50,60 s)of HIFU in vitro to obtain the dosage-effect curves.Then the survival pmtoscolices were incubated,and the mortality of each group was counted daily.The protoscolicidal effects were investigated by trypan blue exclusion assay.Results Compared with the untreated group,the Vitality of E.granulosus protoscolices significantly decreased immediately after treated by HIFU of different dosage(F=5201.59 vs 1865.65,P＜0.05),there were the interaction both different dosage and time(F=214.50,P＜0.05).The protoscolices were broken into pieces by HIFU of 250 W×40 s,whereas the growth of the surviving protoscolices after exposed to HIFU was obvious suppressed.Both the acute killing effect and the delayed inhibitory effect showed a dosage-dependant manner.The inhibitory effect increased along with the increased dosage of HIFU(P＜0.05).The inhibitory effect in 50 W×10 s group was stronger than 25 W×20 s group(P＜0.05).The mortality was increased in parallel with the increase of HIFU dosage.Conclusions HIFU show an effective immediately killing effect,as well as a growth-inhibiting effect on the E.granulosus protoscolices in vitro.

Objective To analyze the treatment outcome and related influencing factors of Tibet- an nationality new smear-positive pulmonary tuberculosis patients in Qinghai Province，so as to provide evidence for tuberculosis control and treatment among Tibetan population． Methods Statistical analysis was conducted on 5 564 Tibetan nationality new smear-positive pulmonary tuberculosis cases in Qinghai province who were reported in the China Tuberculosis Information Management System and approved to receive treatment from 2008 to 2017． The main influencing factors were detected by unconditional Logistic regression model analysis，dependent variable was successful treatment or not，independent variables were other factors related to the treatment outcome． Ｒesults The treatment success rate of Tibetan nationality new smear-positive pulmonary tuberculosis cases was 87. 1% ( 4 848 /5 564) ，and the adverse outcome rate was 12. 9% ( 716 /5 564) ． Unconditional Logistic regression model analysis indicated that non-full- course supervision management，living in agricultural and pastoral area，having severe disease，floating population，and age older than 60 years were risk factors of adverse outcome． The odds ratio( OＲ) 95% confidence interval( CI) of the above risk factors were 13. 044( 10. 671-15. 944) ，2. 305( 1. 703-3. 119) ， 2. 090( 1. 346-3. 243) ，1. 967( 1. 443-2. 682) ，and 1. 909( 1. 410-2. 586) ． Clinical consultation，farmers and herdsmen were protective factors． The OＲ( 95% CI) were 0. 451( 0. 375-0. 543) ，and 0. 786( 0. 627- 0. 985) ． Conclusions Treatment success rate of Tibetan nationality new smear positive pulmonary tuberculosis cases was low． Therefore，the directly observed treatment short-course ( DOTS) strategy should be strictly implemented and the full-course supervision management should be strengthened to improve the treatment success rate． More attention should be paid to the elderly，severe，floating，agricultural and pastoral populations among the Tibetan population．

OBJECTIVETo explore the norms of treatment of acute organophosphorus pesticide poisoning (AOPP), and observe the curative effect.

METHODSOn basis of the pre-research, the norms of treatment of AOPP were summarized, and a multi-center clinical trial was performed in 6 hospitals selected from high incidence of AOPP in Shandong Province.

RESULTS422 patients of AOPP in 6 hospitals in observation period were treated and observed by the norms of treatment. Among them, the proportion of oral poisoning was 97.16%, middle and severe degree were 87.44%. Compared with themselves 2 years ago before standard treatment, the curative effect of the norms of treatment for AOPP was much better than before. The mortality rate of AOPP declined from 9.87% to 1.66% (Chi2 = 27.92, P < 0.01), that was much better than the average therapeutic effect level of all our province in the same period (the mortality rate: 8.92%) (Chi2 = 26.05, P < 0.01). The average amount of atropine [(37.54 +/- 17.76) mg], dropped greatly [(1280.70 +/- 69.22) mg] (U = 439.22, P < 0.01).The usage of atropine by continuous intravenous injection with venous pump was better than ordinary intravenous injection. The mean dosage of pralidoxime chloride increased twice than the previous (U = 19.48, P < 0.01). There was no drug poisoning.

CONCLUSIONThe standard treatment of AOPP is urgently needed in our country, especially in rural area. By this trial, the satisfactory effect of the norms of treatment for AOPP summarized is observed and it reduces the fatality rate remarkably.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Male ; Middle Aged ; Organophosphate Poisoning ; Pesticides ; poisoning ; Poisoning ; therapy ; Standard of Care ; standards ; Young Adult

Adult ; Carcinoma ; virology ; Cervical Intraepithelial Neoplasia ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; In Situ Hybridization ; Middle Aged ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; virology

Mg2Ni0.8Cr0.2-x wt.% CoO/Al2O3 (x=0.5, 1, 2 and 3) composites were prepared by mechanically milling sintered Mg2Ni0.8Cr0.2 alloy and CoO/Al2O3 compound for 45 h. The addition of CoO/Al2O3 compound resulted in the good kinetics properties of hydriding/dehydriding reaction of the composites. The composite with 1.0 wt.% CoO/Al2O3 catalyst could reach the maximum hydrogen absorption capacity (2.9 wt.%) within 5 min at 393 K under H2 pressure of 4 MPa, and can desorb rapidly at 493 K. The decomposition and synthesis of hydrogen molecule on Mg2Ni0.8Cr0.2 alloy surface was promoted by addition of CoO/Al2O3 catalyst. In addition, the formation of metallic Ni particles, strain and defects during the ball milling process also resulted in the improved hydrogenation performance of Mg2Ni-based alloys.
Absorption ; Aluminum Oxide ; analysis ; chemistry ; Hot Temperature ; Hydrogen ; analysis ; chemistry ; Kinetics ; Magnesium Compounds ; analysis ; chemistry ; Manufactured Materials ; analysis ; Materials Testing ; Molecular Conformation ; Nitrogen Compounds ; analysis ; chemistry ; Pressure ; Temperature

A digital image analysis algorithm based color and morphological features was developed to identify the six varieties (ey7954, syz3, xs11, xy5968, xy9308, z903) rice seeds which are widely planted in Zhejiang Province. Seven color and fourteen morphological features were used for discriminant analysis. Two hundred and forty kernels used as the training data set and sixty kernels as the test data set in the neural network used to identify rice seed varieties. When the model was tested on the test data set, the identification accuracies were 90.00%, 88.00%, 95.00%, 82.00%, 74.00%, 80.00% for ey7954, syz3, xs11, xy5968, xy9308, z903 respectively.
Algorithms ; Artificial Intelligence ; Cluster Analysis ; Colorimetry ; methods ; Image Enhancement ; methods ; Image Interpretation, Computer-Assisted ; methods ; Information Storage and Retrieval ; methods ; Oryza ; anatomy & histology ; classification ; Pattern Recognition, Automated ; methods ; Photography ; methods ; Reproducibility of Results ; Seeds ; anatomy & histology ; classification ; Sensitivity and Specificity ; Species Specificity