The ribosomal DNA fragment was amplified with PCR utilizing synthetic primers with host bacteria chromosomal DNA as template. The resultant probes were labeled with Digoxigenin and were applied in the dot blot of DNA vaccine samples. The host bacteria genomic DNA in DNA vaccine against hepatitis B was less than 15 ng/?g. Digoxigenin labeled probes proved sensitive and reliable in determining genomic DNA.

Objective: To investigate the structural characteristics and the primary functions of antigen gene cC1. Meth-ods: The primary and the secondary structures were analyzed using bioinformatics programs provided by Internet servers.The predicted 3D structure of cC1 was established by homology protein modeling method. Anticoagulant activity of GST-anx32 was assayed by modified kaolin partial thromboplastin time (KPTT). Results: cC1 had high homology to annexinsgenes both at nucleic acid and at amino acid level. It contained 4 homologous regions, and each region included the typical an-nexin motif "G-X-G-T (38 residues)-D/E". The results of KPTT assay showed that the recombinant protein GST-anx32 hadhigh anticoagulant activity. Conclusion: cC1 has the common structures of annexins but the homology to the extant annexinsis no more than 48%, cC1 is a member of a novel annexin subfamily and designed as annexin32.

Objective:To identify the HLA-A0201 restricted CTL epitopes derived from hepatitis B virus X protein predicted by computer program and general principles in vitro.Methods:HBx gene sequences of Hepatitis B virus genotypes B/C and serotypes adw/adr,with the highest frequencies in Chinese,were computed and analyzed by screening service offered by Internet combined with peptide supermotif,extended motif and quantitative motif prediction.Four most ideal nine-peptides(HBx1,HBx2,HBx3,and HBx4)were selected as candidate peptides.Using flow cytometry,the fluorescence index of both control and experimental groups were detected and the 4 nine-peptides were evaluated with T2 binding assay and DC50 assay.Results:The nine-peptides VLCLRPVGA(HBx1),CLFKDWEEL(HBx2),VLHKRTLGL(HBx3)and HLSLRGLPV(HBx4)were selected as candidate targets.Among the 4 candidate peptides,HBx2 showed higher HLA-A0201 affinity and HBx2,HBx4 showed better stability.Conclusion:Our study indicates that CLFKDWEEL might be a potential HLA-A0201 restricted CTL epitope from hepatitis B virus X protein;further study is needed for verification of its immunity in vivo.

Objective:To explore the effects of infant sleep patterns on maternal sleep quality.Methods:Three hundred and nineteen pairs of mothers and infants from two grade three class-a hospitals in Nantong City were enrolled. Infant sleep patterns were assessed by the Brief Infant Sleep Questionnaire (BISQ). Maternal sleep quality was evaluated using the Pittsburgh sleep quality index (PSQI). The effect of infant sleep patterns on maternal sleep quality was analyzed by one-way ANOVA followed with multivariate analysis.Results:The maternal sleep quality scored 7.08±2.60. Single factor analysis pointed out that the sleep quality of women whose infants slept in bed near parents, woke up more than 3 times a night, was nocturnal wakefulness more than 60 minutes or nocturnal sleep duration less than 8 hours was significantly worse. After controlling for confounding variables, the way of baby falling asleep, numbers of night wakings, nocturnal sleep duration, maternal age, infant′s gender and primary caregiver were independent factors of maternal sleep quality.Conclusion:Infant sleep pattern is an important factor affecting maternal sleep quality. Nurses should provide information about the sleep patterns of infants to mothers and their family members to promote their babies′ sleep habits so as to improve the maternal sleep quality.

Objective To analyze the correlation between emphysema extent measured by high resolution computed tomography (HRCT) and pulmonary function tests, symptom score in patients with chronic obstructive pulmonary disease ( COPD ) , and to study the value of HRCT in the emphysema quantification in the clinical evaluation of COPD patients. Methods 78 patients with stable COPD were recruited to take the HRCT scan , and emphysema extent was qualified by measuring the proportion of low attenuation area in the whole lung (LAA%). Correlations between LAA% and indices of pulmonary function test, bronchial dilation test, mMRC scale, CAT score and six minutes walking distance (6MWD) were assessed. Results LAA% was negatively correlated with FEV1/FVC and DLCO%pred, and the correlation coefficients were -0.759 and -0.589 (P <0.01), respectively. LAA% was positively related to mMRC score (r = 0.342, P < 0.01), and negatively asso-ciated with 6MWD (r = -0.365,P< 0.01). There was no association between LAA% and indices of bronchodila-tion test (⊿FVC, ⊿FVC%, ⊿FEV1, ⊿FEV1%) (P > 0.05). Conclusions The severity of emphysema measured by HRCT is well correlated with the clinical symptoms , pulmonary function tests and exercise capacity in COPD patients. It can be used to diagnose emphysema early and to evaluate the severity of the disease com-prehensively. Thus, the risk factors of COPD can be controlled and the prognosis of the patients can be im-proved.

Objective: To analyze the strengths, weaknesses, opportunities and threats of the construction on intelligent vaccination clinics in Zhejiang Province, so as to provide countermeasures for promoting the construction of intelligent vaccination clinics in Zhejiang Province. Methods: By reviewing the annual reports of Zhejiang immunization planning, survey data from Zhejiang Centers for Disease Control and Prevention and Immunization Intelligent Service System, data of human resources of immunization planning, vaccine procurement, construction progress of intelligent vaccination clinics and vaccination were collected. The relevant literature was searched to gather information on the construction standards and norms of intelligent vaccination clinics. The analysis of the strengths, weaknesses, opportunities and threats (SWOT) of the construction of intelligent vaccination clinics was conducted, and corresponding countermeasures and suggestions were proposed. Results: The National Immunization Program reported vaccine rate in Zhejiang Province is more than 99%, and standardized vaccination clinics have been popularized throughout the province. The vaccination staff are professional, and a province-wide intelligent immunization service information system has been established, providing the resources and conditions for the construction of intelligent vaccination clinics. However, there are problems such as low data quality and matching efficiency in vaccination, insufficient data interoperability and sharing, unbalanced regional capabilities in intelligent transformation, and uneven distribution of talent and resources. It is crucial to seize the opportunities presented by the development of big data and artificial intelligence, rely on the regional development of the Internet and health industry, seize the opportunity of rapid growth in demand for intelligent vaccination services and high public acceptance, accelerate the construction of intelligent vaccination clinics, and establish intelligent vaccination service standards as soon as possible. Conclusion We should seize the opportunities presented by the digital reform and development, fully utilize the existing vaccination resources and strengths, address the shortcomings, and accelerate the construction of intelligent vaccination clinics in Zhejiang Province.

Objective: o explore the problems encountered in the application of GBZ/T 237-2011 edition of "Diagnosis of Chronic Obstructive Pulmonary Disease Caused by Occupational Stimulating Chemicals" , and to provide reference for the revision of the new standard. Methods: Delphi expert survey method was used to consult experts on the reasonableness of six primary indicators (scope of application, diagnostic principles, diagnostic indicators, treatment principles, diagnostic grading, appendix A) and their corresponding secondary indicators in the Diagnosis of Chronic Obstructive Pulmonary Disease (COPD) caused by Occupational Stimulant Chemicals. Results: The results of the first round of Delphi method showed that the reasonable mean of the diagnostic criteria about long-term occupational history, smoking history and clinical diagnostic criteria was less than 7, and the coefficient of variation was greater than 0.25, indicating that the experts disagreed with the scope of application of the original criteria; the average of smoking history and 3-year working age were 1.78 and 3.43 (less than 7) respectively, which indicated that the experts agreed that the two diagnostic indexes were not conducive to the practical work of occupational disease clinicians; among the diagnostic indicators, smoking history and smoking volume were taken into account in a comprehensive way, and the average values were 7.61 and 7.61 (greater than 7) , respectively. the coefficient of variation was less than 0.25, indicating that the experts had concentrated their opinions and agreed that such indicators could be considered into the diagnostic index system. Conclusion "Diagnosis of Chronic Obstructive Pulmonary Disease Caused by Occupational Stimulating Chemicals" should be improved by expanding the scope of occupational irritant gases. Smoking problems can be considered in combination with clinical practice, and occupational exposure history can be considered to reduce appropriately.

Objective:To explore the effect of postpartum fatigue(PPF) on maternal behavior in rats and its mechanisms.Methods:Sprague-Dawley (SD) rats on the first day after delivery were randomized into the control group and the PPF group using the random number table method, with eight rats in each group.The rat model of PPF was established by forcing rats to stand in a cage with water and last for seven days.To maintain galactosis and lactation, rats and pups were caged for 90 min after every 3 h of separation.The control group was separated routinely without any stimulus.The length and body mass of the pups were recorded at birth and postnatal day 7.On the seven days after modeling, the following maternal behaviors were observed via video recordings: suckling, nesting, clicking and retrieval.The morphology of neurons in the paraventricular nucleus of hypothalamus (PVH) was observed by HE staining.The expression of oxytocin in the paraventricular hypothalamus (OxtPVH) was determined by immunofluorescence and immunohistochemistry.Western blot and qRT-PCR were performed to detect mRNA and protein expression of prolactin (PRL) in pituitary gland, respectively.Statistical analysis was performed by SPSS version 22.0, normally distributed continuous variables were compared between the two groups using an independent-sample t test, and nonnormally distributed continuous variables were compared between the two groups by Mann-Whitney U test. Results:On the seventh day after modeling, the length and weight gain of pups in the PPF group ((5.82±0.17) cm, (5.33±2.54) g)were significantly lower than those of the control group ((6.24±0.36) cm, (7.92±2.54) g, t=3.199, 2.227, both P<0.05). Compared with the control group, the rats in PPF group exhibited abnormal maternal behaviors, such as gnawing cage, biting tails, turning circles, repeatedly nesting and refusal to suckling.The results from the maternal behavioral test revealed that the latency of first pup retrieval and last pup retrieval ((39.25±3.50) s, (280.75±59.16) s) in the PPF group were significantly prolonged compared with those in the control group((19.25±7.68) s, (146.00±49.62) s, t=-4.742, -3.490, both P<0.05), the duration of nesting building ((19.50±12.69) s)and clicking ((95.50±70.55)s) in the PPF group were significantly shorter than those in the control group((68.00±37.59) s, (243.00±62.07) s; t=2.445, 3.139, both P<0.05). Compared with control group, the neurons cells of PVH in the PPF group were in disordered manner and the OxtPVH content in the PPF group decreased significantly.The mRNA (0.33(0.29, 0.38) vs 0.85(0.76, 1.76), Z=-3.576, P<0.05) and protein ((1.00±0.65) vs (4.17±0.49), t=-7.726, P<0.05) levels of PRL in PPF group were significantly decreased compared with those in the control group. Conclusion:The behaviors of holding back, nesting and licking offspring are decreased in postnatal fatigued rats.This may be related to the decreased expression of OxtPVH and PRL in hypothalamus of female rats.

During the occurrence, development and treatment of gynecological tumors, mainly including cervical cancer, endometrial cancer and ovarian cancer, patients are prone to sleep disorders which seriously affect the quality of life. At present, there are few relevant studies on gynecological tumors associated with sleep disorders, and there is a lack of research on TCM syndromes, so there is no targeted treatment plan. This article mainly introduces the present situation of diagnosis and treatment of sleep disorders caused by gynecological tumors in Chinese and western medicine, to provide reference for further study.

Objective To compare the pathological status of gastric mucosa and the expression of HH-PTCH-SMO-GLI(Hedgehog signaling pathway)and NOX/NF-κB/STAT1 signaling pathways in Hp and non-HP infected CAG patients,and to explore the biological mechanism of Hp promoting the"inflammatory cancer transformation"of CAG.Methods 43 patients with CAG who met the criteria were enrolled and divided into CAG with Hp infection group(Hp+ CAG group,n=21)and CAG without Hp infection group(HP-CAG group,n=22).The histological changes of gastric mucosa were observed by hematoxylin-eosin(HE)staining.Western blot was used to detect the relative expression levels of NOX1,NOX2,NOX4,STAT1,P65 and P-P65 in gastric mucosa.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect Gli1 mRNA,Gli2 mRNA,Gli3 mRNA,Shh mRNA,Smo mRNA,Ptch mRNA,NOX1 mRNA,NOX2 mRNA,NOX4 mRNA and NF-κB mRNA in gastric mucosa The mRNA level.Results HE staining results of gastric tissues in the two groups:In the Hp+CAG group,gastric epithelial cells were partially necrotic and shed,the surface was not smooth,the number of glands was reduced and disordered,intestinal metaplasia was observed,and diffuse lymphocyte and neutrophil infiltration were observed in the lamina proper.The degree of lymphocyte and neutrophil infiltration in HP-CAG group was lighter than that in Hp+CAG group.RT-qPCR results:Compared with HP-CAG group,the levels of Gli1 mRNA,Shh mRNA,Smo mRNA and Ptch mRNA in gastric mucosa of Hp+CAG group were significantly decreased(P<0.01).The levels of Gli2 mRNA,Gli3 mRNA,NOX1 mRNA,NOX2 mRNA,NOX4 mRNA and NF-κB mRNA were significantly increased(P<0.01).Western blot detection results:Compared with hP-CAG group,the relative expression levels of NOX1/GAPDH,NOX2/GAPDH,NOX4/GAPDH and P-P65/GAPDH in gastric mucosa of Hp+CAG group were significantly increased(P<0.01),and the STAT1 level was significantly decreased(P<0.01).There was no significant difference in the relative expression of P65/GAPDH between the two groups(P>0.05).Conclusion Hp infection may cause long-term inflammation of gastric mucosa,promote atrophy and intestinal metaplasia,and increase the risk of cancer by inhibiting hH-PTC-SMO-GLi signaling pathway and abnormal activation of NOX/NF-κB/STAT1 signaling pathway.