Humans ; Hypertension, Pulmonary ; etiology ; Male ; Middle Aged ; POEMS Syndrome ; complications

Altitude ; Animals ; Astrocytes ; cytology ; Brain ; cytology ; Hypoxia ; Male ; Microglia ; cytology ; Rats ; Rats, Wistar

Objective:Expression of protein TSLP and selection of full human anti-TSLP single chain Fv ( scFv).Methods:The cDNA of TSLP was amplified.The amplified target gene and the expression vector pET 101/D-TOPO were ligated , and then transformed into E.coli BL21.The protein was induced to expression by IPTG and purified and identified.The biotinylated TSLP protein was used as antigen to select of human TSLP scFv from a constructed human scFv library by phage display .Results: The size of amplified cDNA of TSLP was about 423 bp,and that of expressed protein was about 26 kD.Dot blot and Western blot results showed that the expressed protein was correct.The constructed human scFv library was enriched for three rounds using biotinylated TSLP as antigen by phage display.ELISA results showed that 35% scFvs had binding activity with TSLP.The scFvs with good binding activity were selected and identified by Western blot and sequencing.Conclusion: The full human scFvs against for TSLP were selected suc-cessfully.

Objective:To develop non-immunized human phage display library.Methods:The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:The developed phage library is diversity and can be used for selecting humanized scFv.

Objective:To evaluate the effect of repeated intranasal insulin on postoperative delirium (POD) in elderly patients undergoing general anesthesia.Methods:Seventy elderly patients, aged ≥65 yr, with body mass index ≤28 kg/m 2, of American Society of Anesthesiologists physical statusⅠ-Ⅲ, undergoing elective radical gastrectomy for gastric cancer under general anesthesia, were divided into 2 groups ( n=35 each) according to the random number table method: control group (group C) and insulin group (group I). In group C, normal saline 0.5 ml was administered intranasally twice a day from 2 days before surgery until the day of surgery.In group I, insulin 20 U (0.5 ml) was administered intranasally twice a day from 2 days before surgery until the day of surgery.The regional tissue oxygen saturation (rSO 2) was measured after entering the operating room (T 0), at intubation (T 1), at 1, 2 and 3 h after the start of operation (T 2-4), at the end of surgery (T 5) and at extubation (T 6). The insulin allergic reactions, nasal irritation and hypoglycemic reactions were recorded after intranasal administration of insulin or normal saline within 2 days before operation.The blood glucose concentrations were measured at T 0, T 3 and T 5.The occurrence of POD within 3 days after operation was recorded. Results:Compared with group C, the incidence of POD was significantly decreased within 3 days after operation, and the rSO 2 was increased at T 1-6 in group I ( P<0.05). The rSO 2 was significantly higher at T 1-6 than at T 0 in two groups ( P<0.05). There were no significant changes in blood glucose concentrations at T 0, T 3 and T 5 between the two groups ( P>0.05). No insulin allergic reactions, nasal irritation and hypoglycemic reactions occurred in two groups ( P>0.05). Conclusion:Repeated intranasal administration of insulin can increase the rSO 2 during operation and decrease the occurrence of POD in elderly patients undergoing radical gastrectomy for gastric cancer.

The author introduces the basic concepts of microsatellite and population genetics and its characteristics, expounds the application of these theories for population genetic structure and genetic diversity, gene flow and evolutionary significant unit ESU division research. This paper discuss its applicationin study of genetic causes, origin of cultivation, different regional origins of geoherbs, aiming at providing a new theory and method for geoherbs.
Drugs, Chinese Herbal ; Evolution, Molecular ; Genetic Markers ; genetics ; Genetic Techniques ; Geography ; Microsatellite Repeats ; genetics

OBJECTIVETo explore the impacts on the cognitive level of the children with mental retardation (MR) treated with JIN's three scalp needling therapy and the training for cognitive and perceptual disturbance so as to seek the more effective therapeutic method for the treatment of MR in children. Methods Sixty-six MR children were randomized into a head-point needle-retaining therapy + training group (group A) and a training after acupuncture group (group B). Seven cases and 12 cases were dropped in the two groups respectively. Twenty-six cases accomplished the treatment in the group A and 21 cases in the group B. In the group A, the points on the head were stimulated and the needles were retained (30 min after the general acupuncture, the needles on the body points were removed; and those on the head points were retained for 1 h, including Sishenzhen, Naosanzhen, Zhisanzhen and Niesanzhen). Simultaneously, the training for the cognitive perceptual disturbance was combined. In the group B, 1 h after needle retaining at the body points and head points, all the needles were removed; the training for the cognitive perceptual disturbance was followed. The treatment was given once a day, and the treatment for 3 months was required. Before and after treatment, the Wechsler intelligence scale for children (WISC) was used for evaluation and observation of verbal intelligence quotient (VIQ) , performance intelligence quotient (PIQ) and full-scale intelligence quotient (TFIQ) and score changes of 11 items such as information, picture vocabu; lary, arithmetic, picture generalization, comprehension, etc.

RESULTSAfter treatment, FIQ, VIQ and PIQ scores were different significantly as compared with those before treatment (all P<0. 01). In the group A, the results of picture vocabulary, animal egg laying, maze, block design and geometric figure were all improved significantly (all P<0. 05). In the group B, the results of information, comprehension, block design and geometric figure were all improved significantly (all P<0. 05). After treatment, concerning the value difference in FIQ and PIQ of the two groups; the changes in the group A were more significant (both P<0. 01). After treatment, the results of picture vocabulary and maze were improved significantly in the group A as compared with the group B (both P<. 01). The IQ categories changed apparently after treatment in the two groups, toward the higher level in tendency generally, but without significant difference (both P>0. 05).

CONCLUSIONThe simultaneous treatment with head point retaining of JIN's three needling therapy and the training for cognitive and perceptual disturbance obviously improves children patients' verbal comprehension, expression ability, hand-eye coordination ability, attention, logical reasoning ability and visual perception. The efficacy is better than that in the treatment of the training after acupuncture.

Acupuncture Therapy ; Child ; Child, Preschool ; Cognition ; Female ; Humans ; Intellectual Disability ; psychology ; therapy ; Intelligence Tests ; Male ; Perception ; Scalp ; Treatment Outcome

Molecular identification of Chinese traditional medicine has come from laboratory research into application, but there are some misunderstandings and problems emerging after rapid development. In this paper, we discuss the usage principle, hot field and technology innovation in molecular identification of Chinese traditional medicine. And molecular identification of traditional Chinese medicine has scientific and objective basis, follows the certain systematic research background, and adopts practical principles to establish case by case multi-class identification system. In order to achieve rapid, on-site, high throughput, low cost of traditional Chinese medicine identification purpose, molecular identification technology is further developing for meet the actual needs and the laboratory results further transformation in the service of traditional Chinese medicine industry.
China ; Drugs, Chinese Herbal ; chemistry ; standards ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; classification ; genetics ; Quality Control

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Salvia miltiorrhiza ; enzymology ; genetics

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Bone Marrow ; metabolism ; Brain ; metabolism ; Brain Neoplasms ; metabolism ; pathology ; CDC2 Protein Kinase ; metabolism ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Glioma ; classification ; metabolism ; pathology ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; Young Adult