Objective To investigate the type of plasmid map of Y. pestis in the R. opimas natural plague loci in Junggar Basin. Methods A total of 39 plasmid DNA of Y. pestis which were isolated from the natural plague loci of Junggar Basin, Tianshan Mountain, Kunlun Mountain, Qinghai-Tibet Plateau and In-ner Mongolia were extracted by the methods of Kado and Liu. The plasmid map was analyzed by the methods of agarose gel eleetrophoretogram. Results Two types of plasmid map were found in 26 Y. pestis which were isolated from Junggar Basin. Of them 23 were 6 × 106, 45 × 106 and 65 × 106 type of plasmid map, and 3 were 6 × 106, 45 × 106 and 72 × 106 type. Conclusion There are two types of plasmid map in the R. opi-mus natural plague loci in Junggar Basin. One type, which is the dominant type in this area, is 6 × 106, 45 × 106 and 65 × 106 type. This type is also similar to the dominant plasmid map type of the nature plague loci of Tianshan Mountain, Kunlun Mountain, Qinghai-Tibet Plateau and Inner Mongolia. The other type is 6 × 106, 45 × 106 and 72 × 106 type, and this type is new plasmid map type of Y. pestis in our country.

At present, there is no cure for acquired immune deficiency syndrome (AIDS) due to HIV-1 latent reservoirs. Therefore, it urgently requires novel HIV-1 latency-regulating agents with high potency, low toxicity and favorable drug-like properties to achieve a functional cure for AIDS. Herein, we reviewed the advances in HIV-1 latency-regulating agents since 2019, including the drug discovery strategies, bioactivities, and mechanisms of these compounds. It is of great guiding significance in the development of latency-regulating agents with clinical value.

Objective To evaluate CT and MRI for the diagnosis of cranial extradural empyema.Methods The imaging features in 4 patients with cranial extradural empyema were analyzed.Results 2 cases in frontal,1 case in frontalparietal,1 case in posterier cranial fossa,in this series of 4 cranial extradural empyemas was found homogenous enhancement of dural,and thickened meninges surrounding the empyema.In the series of 1 case show bony thickening and thin.Conclusion The CT and MR of cranial extradural empyema can well demonstrate the morphological and pathological evidence of ivolved menings.Therefore,CT and MR is the most diagnostic value in cranial extradural empyema.

Objective According to principle of forming primer dimer,a new cloning gene method was established. Methods Two and more oligonucleotide with complementary partnership in the 3′ end were synthesized by artifical method,oligonucleotide,Taq DNA polymerase and dNTP were mixed in the proportion of reasonable,then expanded and extended by PCR,result was identifed by agarose gel electrophoresis.Results A clear special band was appeared in the agarose gel. Conclusion Short fragment target gene less than 500 bp may be synthesized by this method.

To improve 3-ketosteroid-△1-dehydrogenase(KSDH) activity and the transformation level for androst-4-ene-3,17-dione, 3-ketosteroid-△1-dehydrogenase gene(ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110?0.5mU and 15?0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

To improve 3-ketosteroid-△1-dehydrogenase (KSDH) activity and the transformation level for androst-4-ene-3,17-dione,3-ketosteroid-△1 -dehydrogenase gene (ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110 ± 0.5mU and 15 ± 0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B.subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

OBJECTIVEThe shear fracture strengths of carbon fiber post, IPS-Empress 2 all-ceramic post (without ZrO2 post), cast Ni-Cr alloy post, prefabricated zirconia ceramic post, human dentin and the shear bonding strengths of the first three kinds of post bonded in the human root canals were measured and compared, which are as the reference for dental clinic.

METHODSStandard cylindrical samples were made respectively in each group of carbon fiber post, IPS-Empress 2 all-ceramic post, cast Ni-Cr alloy post, prefabricated zirconia ceramic post, human dentin, three for each group. The shear fracture strengths of these samples were measured by universal testing machine (AUTOGRAPH DCS5000). Standard cylindrical samples were made respectively in each group of carbon fiber post, IPS-Empress 2 all-ceramic post (without ZrO2 post), cast Ni-Cr alloy post, five for each group. They were bonded in extracted human root canals that had been prepared to 3.0 mm length and 2.0 mm diameter with Glass ionomer cement (Japan Shofu). The shear bond strengths were measured by the same universal testing machine.

RESULTSThe shear fracture strengths of the carbon fiber post (199MPa), cast Ni-Cr alloy post (210MPa, shear bend strength) and prefabricated zirconia ceramic post (193MPa) were statistically higher than those of IPS-Empress 2 all-ceramic post (109MPa) and human dentin (100MPa). The shear fracture strength of the carbon fiber post was statistically similar to that of cast Ni-Cr alloy post and prefabricated zirconia ceramic post. There were no statistical differences between the shear bond strengths of carbon fiber post (2.4MPa) and cast Ni-Cr alloy post (3.8MPa). IPS-Empress 2 all-ceramic post broke before debonding (2.7, break value).

CONCLUSIONSCarbon fiber post, as well as cast Ni-Cr alloy post and prefabricated zirconia ceramic post, has a comparatively high shear fracture strength. The shear bond strengths of carbon fiber post is similar to cast Ni-Cr alloy post.

Case recruitment of large-scale clinical trials should be strictly checked in quality and quantity for it is the key to clinical trial. This study discusses the main difficulties and countermeasures in the case recruitment of large sample, multi-center clinical trials according to the national research project "Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine".

Objective To develop a candidate reference method for the measurement of progesterone in human serum.Methods The serum sample is mixed with the internal standard [3,4-~(13)C_2] progesterone.After extraction with n-hexane and purified by a aqueous solution of 2-Hydroxypropyl-?- cyclodextrin (HP-?-CD),the serum progesterone and labeled progesterone are converted to the 3-enol heptafluorobutyrate and analyzed by gas chromatography mass spectrometry (GC/MS) with selected ion monitoring.The concentration of serum progesterone is calculated by bracketing method.Results The results gave coefficients of variation (CVs) of 0.69% to 2.12%.The analytical recoveries ranged from 98.3% to 100.1%.The results of measuring certified reference materials of serum progesterone are agree with the target value.Conclusion The procedure for measuring progesterone in serum is a highly accurate and precise method and may be used as a candidate reference method for serum progesterone assays.

The selective side-chain cleavage of phytosterol to 4-androstene-3,17-dione(4-AD)and 1,4-androstadiene-3,17-dione(ADD)by Mycobacterium sp.was described.Because of the similarity in chemical structure between 4-AD and ADD,it is difficult to separate them from the fermentation broth.So far,it has been verified that the ADD can be produced by dehydrogenation of 4-AD.In this reaction,3-Ketosteriod-1-Dehydrogenase(ksdD)plays an important role.The gene knocking out method was used to solve the problem.Partial sequence of ksdD was obtained by PCR which was 631bp in length.Then,a targeting vector pUC19-MK was constructed,which was electroporate into the original strain Mycobacterium neoauru.The method of homologous recombination was used to knock out ksdD gene located in the chromosome of Mycobacterium neoauru.In this way,ksdD would lose its enzyme activity.In the result,5 transformants were screened.The experiments of steroid transformation by the transformants were carried out.The productivity of 4-AD reached 17.52% after 144h,which is 192% higher than the original strain.Meanwhile,the productivity of ADD reached 6.12%,which is 89.9% lower than the original strain.