This study was aimed to show the extraction research of cpDNA in Gentiana straminea Maxim. and G. crassicaulis Duthie ex Burk., which will increase the knowledge about the chloroplast genome sequence characteris-tics, understand its genetic diversity and improve the efficiency of breeding schemes. G. straminea Maxim. and G. crassicaulis Duthie ex Burk. were taken as study objects. The improved sucrose-gradient centrifugation and purifica-tion methods were used to obtain the cpDNA, measure the concentration, detect the OD value, and amplify the ITS2 sequence to verify the nucleus pollution. The results showed that the high quality and purity cpDNA (0.1 - 0.4 μg/10 g) was demonstrated without other pollution after ITS2 sequence amplification, which met to the subsequent effi-cient sequencing of chloroplast genomes. The purity and mass have an important influence on the accuracy and cred-ibility of the cp genome sequencing. It was concluded that this study improved the sucrose-gradient centrifugation and purification with great effect of the purity and mass of cpDNA, and guaranteed the obtaining of complete cpDNA of Gentiana.

Objective To analyze psychological status of the donors and the recipients before and after the relative kidney transplantation.Method Symptom checklist 90 (SCL-90) were performed for 147 renal transplant recipients and donors preoperative and postoperative.Statistical analysis were performed to analyze the scores between the recipients,donors and normal standards.Results Over 90％ recipients were of obvious anxiety preoperative.The scores of most factors of recipients were significantly higher than those of donors.The scores of somatization,interpersonal sensitivity,depression,anxiety,hostility,phobia and paranoia between two groups has statistical significance(P＜0.05).Part of recipients retest SCL-90 3 month after operation,the data showed that the scores of somatization,interpersonal sensitivity,depression,anxiety,hostility were obviously declined after operation(P＜0.05).And the test also showed that most of the donors were willing to help preoperative.Conclusion Preoperative psychological test was useful in preoperational psychological intervention for transplantation recipients.It can increase the safety of the recipients during perioperative period.

OBJECTIVETo study the relationship between SNP rs17401966 at the KIF1B gene and the genetic susceptibility to Hepatocellular carcinoma (HCC).

METHODSAll study objects were recruited from two Grade A hospitals of Amoy from January 2011 to October 2014.They were surveyed in individual matching case-control study. Accepting criterias in the cases: HCC was first diagnosed based on diagnostic basis during the investigations, over 18 years old, present addresses were as same as surveyed areas in the district (county) level range, no past history of cancers; Exclusion criterias: patients with other liver diseases. The tumor patients without HCC, patients with autoimmune hepatitis or toxic hepatitis, patients who refused to be investigated or too ill to be investigated. Accepting criterias in the controls: the control who passed the physical examination matched the case in ages (no more than 3 years old), sex, health screening in the same hospital over the same period and district (county); Exclusion criterias: people with liver disease or any history of cancers. This study consisted of 376 HCC patients and 403 controls, 5 ml morning fasting venous blood of all subjects were obtained to isolate cells and distribute genotype. The differences in general information between cases and controls were tested by χ² test and t-test. The association between SNP rs17401966 and the risk of developing HCC were assessed by using the multiple factors logistic regression.

RESULTSThe mean age and standard deviation for case and control groups were (61.7 ± 12.8) years and (60.6 ± 12.7) years (t = 1.15, P = 0.251), respectively. The proportion of family history of cancer [28.7% (108/376)] and the HBsAg positive rate [26.9 % (101/376)] in case group were higher than these in control group [15.9% (64/403), 2.7% (11/403)] (χ² = 18.65, 92.02, P < 0.001). In HBsAg carriers, GG genotype genetic susceptibility to HCC is 0.12 (0.02-0.75) times for AA genotype, and G allele susceptibility to HCC is 0.38 (0.15-0.98) times for A allelc. In HBsAg negative group, it showed no statistical significance in the relationship between SNP rs17401966 and susceptibility to HCC, and compared with the A allele, the risk for HCC of G allele is 0.79 (0.62-1.01).

CONCLUSIONThe results demonstrated that the presence of the GG genotype, the GA genotype and the G allele at rs17401966 of the KIF1B gene might decrease the risk for HCC.

Aged ; Alleles ; Carcinoma, Hepatocellular ; Case-Control Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Kinesin ; Liver Neoplasms ; Middle Aged ; Polymorphism, Single Nucleotide

Objective To produce human papillomavirus type 6(HPV-6)virus-like particles with Escherichia coli expression system and study its immunogenicity.Methods HPV-6 L1 gene was inserted into pmkaryotic expression vector pTO-T7 and then expressed in Escherichia coli ER2566.The HPV-6 L1 protein was purified by ammonium sulfate precipitation,ion-exchange chromatography,and hydrophobic interaction chromatography.Then the purified HPV-6 L1 self-assembled into virus-like particle after removing 1,4dithiothreitol(DTr).The morphology of the virus-like particles was investigated with dynamic light scatter and transmission electron microscopy,and the immunogenicity was determined with in vitro pseudownons neutralization as8ay.Results HPV-6 L1 was expressed in soluble form in Escherichia coli.Following the removal of DTT,purified HPV-6 L1 protein could assemble into virus-like particles as 25 am in the radius.And the animal immunization test showed HPV-6 virus-like particles can elite hish titer neutralizing antibodies.Conclusion The bacterially expressed HPV-6 L1 VLP is highly immunogenieity and easy to produce.And it can be good candidate of HPV-6 vaccine.

Although previous publications suggest the 2009 pandemic influenza A(H1N1)virus was reassorted from swine viruses of North America and Eurasia, the immediate ancestry still remains elusive due to the big evolutionary distance between the 2009 H1N1 virus and the previously isolated strains. Since the unveiling of the2009 H1N1 influenza, great deal of interest has been drawn to influenza, consequently a large number of influenza virus sequences have been deposited into the public sequence databases. Blast analysis demonstrated that the recently submitted 2007 South Dakota avian influenza virus strains and other North American avian strains contained genetic segments very closely related to the 2009 H1N1 virus, which suggests these avian influenza viruses are very close relatives of the 2009 H1N1 virus. Phylogenetic analyses also indicate that the2009 H1N1 viruses are associated with both avian and swine influenza viruses circulating in North America. Since the migrating wild birds are preferable to pigs as the carrier to spread the influenza viruses across vast distances, it is very likely that birds played an important role in the inter-continental evolution of the 2009 H1N1virus. It is essential to understand the evolutionary route of the emerging influenza virus in order to find a way to prevent further emerging cases. This study suggests the close relationship between 2009 pandemic virus and the North America avian viruses and underscores enhanced surveillance of influenza in birds for understanding the evolution of the 2009 pandemic influenza.

Butanol production from acid hydrolysate of Jerusalem artichoke juice by Clostridium acetobutylicum L7 was investigated, and it was found that natural components of the hydrolysate were suitable for solvent production with the species. With batch fermentation using the medium containing 48.36 g/L total sugars, 8.67 g/L butanol was produced at 60 h, and the ratio of butanol to acetone to ethanol was 0.58:0.36:0.06, which were similar to the fermentation with fructose as carbon source, but both of these two fermentations were slower than that with glucose as carbon source, indicating the fructose transport of the species might not be effective as that for glucose. When the total sugars of the medium were increased to 62.87 g/L, the residual sugars increased slightly from 3.09 g/L to 3.26 g/L, but butanol production of the fermentation system was improved significantly, with 11.21 g/L butanol produced and the ratio of butanol to acetone to ethanol at 0.64:0.29:0.05, which illustrated that an excess in sugars enhanced the butanol biosynthesis of the species by compromising its acetone production. When the sugar concentration of the medium was further increased, much more sugars were remained unconsumed, making the process economically unfavourable.
Butanols ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Helianthus ; chemistry ; Industrial Microbiology ; methods

Objective:To investigate the clinical and biological characteristics of relapsed childhood low-risk acute B lymphoblastic leukemia (B-ALL).Methods:The clinical and laboratory data of 34 children who admitted in Beijing Boren Hospital from July 2017 to July 2018 were retrospectively analyzed, and 127-339 mutations of hematological malignancy related genes were analyzed.Results:The median time from the diagnosis to the recurrence was 871 d (87-1 446 d). The recurrence at early stage and late stage had 26 cases (76%) and 8 cases (24%), respectively. The recurrence before maintenance treatment, during maintenance therapy and after withdrawal of chemotherapy had 3 cases (9%), 12 cases (35%) and 19 cases (56%) (13 cases relapsed within 1 year after withdrawal, 6 cases relapsed after withdrawal 1-2 years and no one relapsed after withdrawal 2 years). The sites of recurrence included bone marrow alone accounting for 26 cases (76%), both intramedullary and extramedullary disease (EMD) accounting for 6 cases (18%), EMD alone accounting for 2 cases (6%). Flow cytometry showed that 9 patients presented minimal residual disease (MRD)-positive (6 cases with one positive, 2 cases with twice positive and 1 case with 3 times positive), including 8 cases occurred at early stage and 1 case occurred at late stage; and the level of MRD was 0.02%-3.82%. Complex chromosomal karyotype appeared in 6 relapsed children with normal or hyperdiploid karyotype at first diagnosis. Hematological malignancy related gene mutation detection was made in 28 cases, and the results showed that each patient had at least one gene mutation, and 2 or more gene mutations were detected in 25 cases (89%). The high frequency of gene mutations were as follows: CREBBP (7 cases, 25%), NRAS (7 cases, 25%), KRAS(7 cases, 25%), TP53 (4 cases, 14%), and NT5C2 (4 cases, 14%).Conclusions:The recurrence of childhood low-risk B-ALL occurs mostly in the maintenance treatment or in two years of withdrawal of chemotherapy. Positive MRD after complete remission is likely to show the risk of early recurrence. The gene mutations after the poor prognosis in cancer cells may be related to the recurrence of childhood low-risk B-ALL, and the common gene mutations include CREBBP, RAS signaling pathways genes and TP53, NT5C2.

Objective:To observe the clinical effect of tranexamic acid radiofrequency introduction combined with Q-switched laser comprehensive treatment of moderate to severe melasma.Methods:From December 2019 to September 2021, ninety-six female patients with melasma [age 24-59 years old, average age (37.8±6.0) years old] were admitted to the Plastic and Aesthetic Department of the First Affiliated Hospital of Henan University of Chinese Medicine. They were divided into the Q-switched laser group and the combined group by random number table method, with 48 cases respectively. The Q-switched laser group received Q-switched laser therapy, while the combined group received tranexamic acid radiofrequency introduction combined with Q-switched laser comprehensive therapy. The melasma area and severity index (MASI) scores were compared between the two groups before and after treatment. The clinical efficacy, adverse reactions and recurrence rates of the two groups were compared.Results:The MASI scores of the Q-switched laser group and the combined treatment group were (28.28±1.24) points and (28.52±4.25) points respectively before treatment, and (13.38±7.96) points and (9.11±5.48) points respectively after treatment. The MASI scores of the two groups were decreased after treatment, which of the combined group was lower than that of the Q-switched laser group ( t=3.06, P<0.05). The total clinical effective rate of the combination group (93.75%) was higher than that of the Q-switched laser group (79.17%) (χ 2=4.36, P<0.05). The incidence rate of hyperpigmentation (2.08%) and recurrence rate (2.08%) of the combination group were lower than those of the Q-switched laser group (14.58%, 16.67%) (χ 2=6.01, P<0.05). Conclusions:Tranexamic acid radiofrequency introduction combined with Q-switched laser comprehensive treatment can improve skin lesions and clinical efficacy in patients with moderate to severe melasma, and reduce pigmentation and recurrence.

Secretory IgA (SIgA) antibodies in external secretions play an important role in mucosal immune response. Polymeric SIgA was advantageous over monomeric IgA (mIgA) and IgG in several aspects. To express secretory IgA antibody against H5N1 virus, we constructed the secretory component and immunoglobulin J expressing plasmids and co-transfected the plasmids into the Chinese hamster ovary cells (CHO) stably expressing immunoglobulin A. Then we used Zeocin to select the positive clone cells, monoclonal cells stably secreting SIgA was screened through fold dilution method at last. The SIgA antibody secreted from the CHO cells was confirmed by Western blotting, which demonstrated that we had got the complete SIgA molecular. The successful expression of this polymeric anti-H5N1 SIgA in CHO cells will contribute to the production of recombinant SIgA as a preventive agent for infectious disease control.
Animals ; Antibodies, Viral ; biosynthesis ; genetics ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Genetic Vectors ; Immunoglobulin A ; immunology ; Immunoglobulin A, Secretory ; biosynthesis ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology

Objective : To investigate the diagnostic endoplasmic reticulum aminopeptidase-1 (ERAP1) in patients with hepatocellular carcinomae (HCC) ． Methods : Enzyme-linked immunosorbent assay (ELISA) was used to de- tect the serum levels of ERAP1 in HCCpatients，cirrhosis patients and healthy controls (HC) ．Multivariate logistic regression was used to analyze the independent risk factors of the severity and prognosis ，and receiver operating characteristic curve (ROC) was used to evaluatesensitivity and specificity of ERAP1 in the diagnosis of different degree of disease and prognosis. Results : The serum ERAP1 level of HCC was related to tumor stage，tumor size and number of cancer focal (P ＜0. 05 ) ． ERAP1 level of HCC patients was positivecorrelated with ALT ，AST， TBIL and AFP，while negative correlated with ALB(P＜0. 05) ．ERAP1 was found to be an independent predictor of different severity and prognosis．When joint diagnosing HCC with AFP，the area under the curve ( AUC) was 0. 932．For the diagnosis of poor prognosis，the AUC was 0. 742． Conclusion Serum ERAP1 level has important clinical significance and potential application value in evaluating the severity and prognosis of HCC patients.