Objective:To investigate the purification of antibacterial effective fraction of Syringae folium by macroporous resins. Methods:Static adsorption and desorption tests were carried out to screen the macroporous resins. The desorption experiment was per-formed on the selected D101 resin to optimize the separation process. The effects of resin amount, diameter length ratio, elution flow rate, elution solution concentration and volume were studied. Results:The optimal conditions were as follows:the elution solution was 55% ethanol, the adsorption flow rate was 1 BV·h-1 , the elution flow rate was 5 BV·h-1 , 6 BV 25% ethanol was used to eliminate impurity and 8 BV 55% ethanol was used to elute to obtain the effective fraction. Conclusion: The content of antibacterial effective component is above 65% after purified by D101 resin, indicating that the present method is suitable for large-scale preparation of anti-bacterial effective fraction of Syringae folium.

Objective To screen the sensing elements for TNT detection in Escherichia coli genome.Methods A genome promoter library with cutting E.coli K-12 MG1655 genome was constructed.Bacterial luciferase luxCDABE was used as a reporter gene during promoter screening.We discovered TNT sensing elements through several rounds of screen-ing.Through analysis of sensitivity, specificity and timeliness, the promoter activity of the elements was evaluated,and the functional sequence of the elements was further confirmed.Results and Conclusion We successfully constructed an E.co-li K-12 MG1655 genome library , from which a TNT sensing element was discovered,which had a good performance in the analysis of sensitivity, specificity and timeliness.In this study, we reported that the topAp4 is a TNT sensing element for the first time.We also verified its excellent promoter activity.

Objective To investigate the clinical application value of the levels of heparin-binding protein (HBP) in cerebrospinal fluid (CSF) for intracranial infectious diseases. Methods A case-control study was conducted. 150 patients after craniotomy(73 in the postoperative bacterial intracranial infection group, 77 in the postoperative non-infection group) admitted to the Department of Neurology of the People's Hospital of Liaoning Province from December 2016 to May 2018 were collected. At the same time, 46 patients without operation (14 in the non-bacterial intracranial infection group, 32 patients without intracranial infection were selected as control group whose white blood cell count (WBC) values in CSF were all below 10 × 106/L) in the same period were also collected. According to the diagnostic criteria for severe intracranial infection, the patients with bacterial intracranial infection were divided into 26 cases of mild intracranial infection group and 47 cases of severe intracranial infection group. According to the Glasgow Outcome Scale (GOS) score at the time of discharge, the patients were divided into 30 cases of good prognosis group (GOS score 4-5 points) and 43 cases of poor prognosis group (GOS score 1-3 points). The concentrations of HBP in CSF were tested with latex immunoturbidimetry, and the concentrations of procalcitonin(PCT) in cerebrospinal fluid and serum were tested with electrochemiluminescence, and cerebrospinal fluid routine were tested with instrument method, and the concentrations of total protein(TP) in cerebrospinal fluid were tested with turbidimetry. The differences of the laboratory test indicators in each group were statistically analyzed, and the levels of HBP in CSF of patients with different degrees of intracranial infection and different prognosis were compared. Comparison of two independent samples was performed using the Mann-Whitney U test. Results The HBP levels in cerebrospinal fluid were 187.00 (73.00, 635.00) ng/ml, 10.00 (3.50, 32.00) ng/ml, 1.50 (0, 4.00) ng/ml, 3.00 (1.00, 4.00) ng/ml in post-craniotomy bacterial intracranial infection group, uninfected group after craniotomy, non-bacterial intracranial infection group and control group respectively. The cerebrospinal fluid levels of WBC count were 1280.00 (363.00, 4327.00)×106/L, 63.00 (18.50, 300.00)×106/L, 5.00 (3.00, 14.75)×106/L, 3.00 (2.00, 5.75)×106/L. The absolute value of cerebrospinal fluid neutrophils were 1216.00 (225.50, 3895.50)×106/L, 24.00 (2.00, 209.50)×106/L, 1.00 (1.00, 3.00)×106/L, 1.00 (1.00, 1.00)×106/L. The cerebrospinal fluid levels of PCT were 0.16 (0.10, 0.32) ng/ml, 0.09 (0.07, 0.14) ng/ml, 0.07 (0.06, 0.12) ng/ml, 0.07 (0.06, 0.13) ng/ml. The serum levels of PCT were 0.36 (0.15, 1.09) ng/ml, 0.09 (0.04, 0.16) ng/ml, 0.08 (0.04, 0.13) ng/ml, 0.07 (0.03, 0.11) ng/ml. The levels of HBP, WBC, neutrophils, PCT in CSF and serum PCT in the post-craniotomy bacterial intracranial infection group were significantly higher than those in the uninfected group after craniotomy (Z=-9.246,-6.759,-6.741,-4.477,-6.202, P<0.05), non-bacterial intracranial infection group(Z=-5.840,-5.412,-5.259,-2.923,-5.104,P<0.05) and the control group (Z=-7.905,-7.919,-7.335,-4.397,-5.474, P<0.05). There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-3.763,-3.444,-3.041,P<0.05) and no significant differences in CSF and serum PCT (Z=- 0.869, - 1.850, P>0.05)between the uninfected group after craniotomy and the non-bacterial intracranial infection group. There were significant differences in the levels of HBP, WBC and neutrophils in CSF(Z=-4.496,-6.685,-4.842,P<0.05) and no significant differences in CSF and serum PCT(Z=-0.676,-1.303, P>0.05)between the uninfected group after craniotomy and the control group. There were no significant differences in the levels of HBP, PCT in CSF and serum PCT (Z=-0.861,-0.514,-0.273, P>0.05)and significant differences in the levels of WBC and neutrophils in CSF(Z=-2.756,-3.060, P<0.05) between the non-bacterial intracranial infection group and the control group. The levels of HBP in CSF in the severe intracranial infection group were significantly higher than those in the mild intracranial infection group(Z=-6.267, P<0.05). The levels of HBP in CSF in the poor prognosis group were significantly higher than those in the good prognosis group(Z=-7.064, P<0.05). The area under the ROC curve for the diagnosis of bacterial intracranial infection by HBP, WBC, neutrophils, TP, PCT in CSF and PCT in serum was 0.986, 0.987, 0.945, 0.945, 0.770 and 0.914, respectively. The area under the ROC curve for differential diagnosis of bacterial intracranial infection and non-bacterial intracranial infection was 0.994, 0.958, 0.961, 0.929, 0.747 and 0.936, respectively. Conclusions HBP in CSF is an ideal indicator for the diagnosis of bacterial intracranial infection. It is important to distinguish between bacterial intracranial infection and non-bacterial intracranial infection. The extent of increase is related to the severity of infection and prognosis of the disease.

Antibiotic-associated diarrhea(AAD) is a frequent adverse effect of antibiotic in children.AAD is associated with longer hospitalization, higher healthcare cost and even lead to death.Pediatricians usually do not pay enough attention to AAD.Domestic experts from pulmonary medicine, infection and gastroenterology are organized to develop the consensus, to improve the diagnosis, treatment and prevention of AAD, and contribute the children health in future.

OBJECTIVETo investigate HIV-1 co-receptor usage in patients experienced anti-retroviral therapy (ART) in Anhui and Henan province of China.

METHODSA total of 45 HIV-1 infected individuals who have experienced ART and 109 un-experienced ART patients from Anhui and Henan province, which were called as treatment group and treatment-negative group, were selected as study subjects. HIV-1 strains were isolated from peripheral blood mononuclear cells of whole blood from patients. HIV-1 p24 in the culture supernatant was measured using a commercial enzyme-linked immunosorbent assay (ELISA) kit. HIV-1 co-receptor usage was identified using Ghost cell lines expressing CD4 and the chemokine receptor CCR5 or CXCR4.

RESULTSAmong 45 HIV strains from the treatment group, 22 (48.9%) strains used CCR5 as a co-receptor (R5 tropic strain), 21 (46.7%) strains used CXCR4/CCR5 as a co-receptor (X4/R5 duel tropic strain), and 2 (4.4%) used only CXCR4 as a co-receptor (X4 tropic strain). In 109 strains from treatment-negative group, 96 (88.1%) strains used CCR5 as a co-receptor (R5 tropic strain), 13 (11.9%) strains used CCR5/CXCR4 as a co-receptor use (X4/R5 strain). A significant difference was found between two groups in X4 co-receptor usages (χ(2) = 27.30, P < 0.05). Furthermore, after treated with AZT + DDI + NVP, the HIV-1 CXC4/CCR5 utilization was 59.09% (13/22), meanwhile after treated with D4T + DDI + NVP, the HIV-1 CXC4/CCR5 utilization was 43.48% (10/23), which the difference was not statistical significant (χ(2) = 1.10, P = 0.30).

CONCLUSIONHIV-1 CXCR4/CCR5 co-receptor utilization was higher in ART patients than treatment-negative patients.

Acquired Immunodeficiency Syndrome ; drug therapy ; metabolism ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Cells, Cultured ; Female ; HIV-1 ; isolation & purification ; Humans ; Male ; Middle Aged ; Receptors, CCR5 ; metabolism ; Receptors, CXCR4 ; metabolism ; Receptors, HIV ; metabolism

Objective To explore the ethnic differences on nutritional status of children under three years old,in the western areas of China and to indentify the affecting factors.Methods 14 072children under the age of three years and their mothers were recruited,using the proportion population sampling method in 45 counties in the western areas of China.Height and weight were used as the measurement on nutrition of children.Results Height and weight of children with Han,Uyghur,Tibetan and Zhuang ethnicities were all lower than the WHO standards and with differences on height and weight among them.There were also ethnic differences on the status of malnutrition.Prevalence rates of stunting were 14.7％,20.3％,26.9％ and 26.5％ for Han,Uyghur,Tibetan and Zhuang ethnicities,respectively.Prevalence rates of underweight were 6.1％,10.7％,6.8％ and 15.5％ among the Han,Uyghur,Tibetan and Zhuang ethnicities,respectively.The prevalence rates of wasting were 4.2％,5.3％,2.9％,8.9％,and of under nutrition were 19.2％,25.5％,30.3％ and 36.5％ for the Han,the Uyghur,the Tibetan and the Zhuang ethnicities,respectively.When factors as family size,years of schooling of parents,family numbers,sex,age (months),mother' s height and weight,ways of feeding and sources of family income etc.were adjusted,children with Uyghur,Tibetan,Zhuang and other ethnicities were still presented more malnutrition than the children with Han ethnicity.Conclusion There were differences on malnutrition status of children under the age of three years among the studies on different ethnicities.Undemutrition was less than nutritional status,seen in children of the Han nationality than other minority ethnicites.Ethnicity seemed to be related to differences in the nutritional status of children in western China while children of the Han nationality would be better than other minorities on nutritional status if they were in same living conditionals.

OBJECTIVETo develop a method for detection of coxsackie B virus type 1-6 by RT-PCR.

METHODSA pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus.

RESULTSThis method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR.

CONCLUSIONThe RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.

DNA Primers ; Enterovirus B, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; diagnosis ; virology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin M ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR.

METHODSPrimers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested.

RESULTSThe positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160).

CONCLUSIONThe simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.

Bodily Secretions ; virology ; DNA Primers ; Humans ; RNA, Viral ; blood ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; SARS Virus ; genetics ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; blood ; diagnosis ; virology

BACKGROUNDProteasome subunits (PSMB) and transporter associated with antigen processing (TAP) loci are located in the human leukocyte antigen (HLA) Class II region play important roles in immune response and protein degradation in neurodegenerative diseases. This study aimed to explore the association between single nucleotide polymorphisms (SNPs) of PSMB and TAP and Parkinson's disease (PD).

METHODSA case-control study was conducted by genotyping SNPs in PSMB8, PSMB9, TAP1, and TAP2 genes in the Chinese population. Subjects included 542 sporadic patients with PD and 674 healthy controls. Nine identified SNPs in PSMB8, PSMB9, TAP1, and TAP2 were genotyped through SNaPshot testing.

RESULTSThe stratified analysis of rs17587 was specially performed on gender. Data revealed that female patients carry a higher frequency of rs17587-G/G versus (A/A + G/A) compared with controls. But there was no significant difference with respect to the genotypic frequencies of the SNPs in PSMB8, TAP1, and TAP2 loci in PD patients.

CONCLUSIONChinese females carrying the rs17587-G/G genotype in PSMB9 may increase a higher risk for PD, but no linkage was found between other SNPs in HLA Class II region and PD.

ATP-Binding Cassette Sub-Family B Member 2 ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; genetics ; Adult ; Aged ; Antigen Presentation ; Case-Control Studies ; Cysteine Endopeptidases ; genetics ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; immunology ; Polymorphism, Single Nucleotide ; Proteasome Endopeptidase Complex ; genetics

OBJECTIVE: To study the clinical and sequence character of the entire mitochondrial genome in five subjects with mitochondrial 12SrRNA T1095C mutation, and to analyze its relationship with the military noise-induced hearing loss (NIHL). METHOD: Three hundreds and four soldiers exposed to military noise were selected in Yunan and Beijing, including susceptible (experimental) and tolerance (control) groups. Mitochondrial 12SrRNA T1095C mutation were found in 5 subjects. Then the complete nucleotide sequence of five subjects were sequenced and its clinical character were analyzed. RESULT: m12SrRNA T1095C mutation were identified in 5 subjects of experimental group,and none were found in control group. There was significant difference between them (P < 0.05). All five soldiers had the history of military noise exposure and showed sensorineural deafness of different degrees. Sequence analysis of the complete mitochondrial genomes showed the distinct sets of mtDNA polymorphism besides T1095C mutation in five subjects. CONCLUSION The T1095C mutation in hearing loss subjects with various genetic background and history of military noise exposure, is involved in the pathogenesis of hearing impairment. It indicates that the T1095C mutation do relate well with military noise induced-hearing loss.
Adult ; Base Sequence ; DNA, Mitochondrial ; genetics ; Hearing Loss, Noise-Induced ; genetics ; Humans ; Male ; Military Personnel ; Mutation ; Young Adult