Objective To establish a duplex fluorescence PCR for detection of HIV proviral DNA and to evaluate its application for early diagnosis of HIV infection in infants .Methods A duplex fluores-cence PCR system was set up based on TaqMan technology for detection of human ribonuclease P ( RNase P) gene and long terminal repeat ( LTR) region of HIV.A recombinant plasmid containing the targeted gene fragment , pTG19-T, was constructed by TA cloning technique and used as the template for evaluation of sen -sitivity of the assay .Blood samples from 11 healthy individuals and 98 HIV-infected patients were collected and detected to validate the assay specificity .The assay of duplex fluorescence PCR was then carried out to detect 96 infant blood samples collected from several maternal and child health hospitals in Zhejiang province from January 2011 to September 2012 for early diagnosis of HIV infection .The results were compared with those by using the Roche HIV DNA qualitative detection kit .Results The established duplex fluorescence PCR could specifically detect HIV proviral DNA with a specificity of 100%and a detection sensitivity of 100 cps per reaction .The coincidence rate between the established assay and the Roche HIV DNA qualitative de -tection kit was 100%in the detection of 96 blood samples .Conclusion The duplex fluorescence PCR as-say showed advantages of cost-effectiveness , convenience , good specificity and accuracy with high sensitivi-ty.It could be used for early diagnosis of HIV infection in infants and also as a general technical platform for the detection of HIV proviral DNA .

OBJECTIVE To survey the distribution of class Ⅰ integron in extended-spectrum beta-lactamases(ESBLs)-producing Escherichia coli and Klebsiella pneumoniae and its contribution in horizontal transfer of ESBLs genes.METHODS The presence of class Ⅰ integron among 230 ESBLs-producers and 197 non-ESBLs-producers of E.coli and K.pneumoniae were detected by PCR.The correlation and co-transfer between integron and genes coding for SHV,CTX and TEM were studied. RESULTS One hundred and thirty-seven(59.6%) isolates were positive for intⅠ gene among ESBLs-producers,contrasted to 48(24.4%) in non-ESBLs-producers(P

Objective:To compare the performance of four commercial kits for anti-HIV confirmatory assay and to provide a reference for clinical application.Methods:Two western blot (WB) kits and two recombinant immunoblot assay/line immune assay kits were used to detect 185 samples in parallel, McNermar test was used to analyze the consistency between the testing results and HIV infection status. Compared to the testing result of the most used MP company’s WB kit, the odds ratio ( OR) was calculated. Results:The consistency of the testing results with the status of HIV infection by the four kits was 83.24%-92.43%, and the Kappa value was 0.558-0.831.The OR value of IMT company’s WB kit obtained from 57 HIV-uninfected samples was 0.932(95% CI: 0.446-1.946, P=0.851), and the OR values of WT company’s recombinant immunoblot assay (RIBA) and MK company’s line immune assay (LIA) were 2.348(95% CI: 1.069-5.158, P=0.034) and 23.064 (95% CI: 5.125-103.789, P<0.001), respectively. The OR value of LIA obtained from 128 HIV-infected samples was 0.153 (95% CI: 0.034-0.700, P=0.016). LIA showed fewest reactions by testing 95 HIV positive screening samples, which was significantly different from any other kit ( P< 0.05). There was no significant difference among the other three kits. In the case of non-specific reactions, the numbers of bands were all ≤4, and the proportion of light-colored bands accounted for 52.1% (37/71). The main bands were gp160(81.3%), p24(71.9%), and p17(28.1%). Conclusions:There was no significant difference between the two WB kits. The RIBA/LIA detection kits could reduce the generation of indeterminate result, but the LIA kit has the risk of false negative for weakly reactive samples.

Objective:To explore the research hotspots of early enteral nutrition and analyze its development trend.Methods:The Web of Science core database was retrieved. HistCite and CiteSpace were used to conduct quantitative analysis and co-word clustering analysis of early enteral nutrition.Results:A total of 823 articles were retrieved, and the number of articles was increasing. The research hotspots of early enteral nutrition mainly included severe disease, esophageal cancer, acute pancreatitis, sepsis, malnourished patients and premature infants. At the same time, the selection of early enteral nutrition nutrients was also a research hotspot.Conclusions:Early enteral nutrition research in critically ill patients is mature, and other specialized fields can carry out specialized early enteral nutrition support based on the research on critically ill patients. In the future, comparative studies on the effects of different nutrients in early enteral nutrition can also be carried out.

Objective:To investigate the characteristics of subtype diversity and transmission on HIV-1 among 12 to 30 years old student MSM in Zhejiang province.Methods:A total of 290 newly diagnosed HIV infected student MSM were selected as the research objects for molecular studies on HIV, in Zhejiang province during 2013 to 2015. Data on epidemiology and plasma samples of these people were collected. HIV-1 nucleotide sequences of pol gene regions were amplified using the RT-PCR/nested PCR method and sequenced. Phylogenetic analysis was performed to determine the HIV-1 genotypes. Characteristics of transmission mode among these cases were also analyzed. Results:A total of 290 cases, 50.3 % were diagnosed in Hangzhou and 81.0 % had college or above degrees. 178 sequences including 10 subtypes, were obtained, with the main subtypes as CRF01_AE (49.4 %, 88/178) and CRF07_BC (39.3 %, 70/178). A total of 18 molecular transmission clusters were formed (42 cases, cluster size from 2 to 4), with the proportions of clusters as 23.6 % (42/178). 61.9 % (26/42) of student MSM with their schools located in the same district within the transmission clusters. Their sexual partners would include both student MSM and non-student MSM. The proportion of clusters among middle school students was 38.2 % (13/34), higher than that of college students (20.1 %, 29/144) ( χ2=4.996, P<0.05). Conclusions:The HIV-1 subtypes of student MSM in Zhejiang province appeared diversity, which indicated with the diversity of sources of infection. The geographical distribution of cluster cases is relatively centralized. In order to effectively control the spread of AIDS, more attention should be paid to the sexual partners involved and to specific programs on intervention.

Objective:To investigate the relationship between HIV-infected men who have sex with men and their sexual partners in Zhejiang province.Methods:A cross-sectional survey was conducted to recruit newly confirmed HIV/AIDS among MSM from 2015 to 2017, including sexual partner identification and molecular epidemiological study. Plasma was collected to extract RNA, and the pol gene of HIV-1 was amplified by RT-PCR/nested PCR. Phylogenetic tree and molecular transmission cluster were analyzed to identify the transmission relationship between sexual partners. Results:A total of 937 HIV/AIDS among MSM were recruited to promote HIV testing for their sexual partners, and 173 positive sexual partners were identified. 50.8% (61/120) of the gene sequences were clustered among the positive sex partners. Seven pairs of clustered sex partners combined with the results of recent infection preliminarily determined the transmission direction. In the clusters, there were statistical differences between the partners who were diagnosed in the same year ( OR=12.190, 95% CI: 1.563-95.054) or with current residence in the different districts ( OR=17.054, 95% CI: 1.742-166.982). Conclusions:Combined with a molecular transmission network, HIV test for the sexual partners of HIV/AIDS among MSM can improve the accurate tracking of cases and preliminarily determine the direction of transmission, according to the results of recent infection. It is suggested that after HIV is confirmed for HIV/AIDS among MSM, HIV tests should be carried out as soon as possible for their sexual partners, including a cross-regional sexual partner tracking test, which is helpful to improve the tracing procedure.

Objective:To explore the effects of acute paraquat poisoning on cognitive function of patients through neuropsychologic test.Methods:In June 2019, 36 patients with acute paraquat poisoning in the emergency department of a provincial hospital in Hebei Province were selected as the case group. 36 healthy individuals were selected as control group. The cognitive function and depressive state were assessed by mini mental state scale, auditory word learning test, digit span test, connection test, Boston Naming Test and geriatric depression scale.Results:The results of Mini-Mental State examination showed that the total score of the case group was lower than that of the control group, the difference was statistically significant ( P<0.05) . The results of the Auditory Vocabulary Learning test showed that the scores of delayed recall, clue recall, corrective ability and semantic learning strategies of the case group were significantly lower than those in the control group ( P<0.05) . There was no significant difference in the scores of immediate memory between the two groups ( P>0.05) . The scores of Digit Span test and Boston Naming test in the control group were higher than those in the case group, the Trail Making test time in the control group was shorter than that in the case group, and the differences were statistically significant ( P<0.05) . Conclusion:Acute paraquat poisoning can impair human cognitive ability to a certain extent.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

Objective:To explore the effects of acute paraquat poisoning on cognitive function of patients through neuropsychologic test.Methods:In June 2019, 36 patients with acute paraquat poisoning in the emergency department of a provincial hospital in Hebei Province were selected as the case group. 36 healthy individuals were selected as control group. The cognitive function and depressive state were assessed by mini mental state scale, auditory word learning test, digit span test, connection test, Boston Naming Test and geriatric depression scale.Results:The results of Mini-Mental State examination showed that the total score of the case group was lower than that of the control group, the difference was statistically significant ( P<0.05) . The results of the Auditory Vocabulary Learning test showed that the scores of delayed recall, clue recall, corrective ability and semantic learning strategies of the case group were significantly lower than those in the control group ( P<0.05) . There was no significant difference in the scores of immediate memory between the two groups ( P>0.05) . The scores of Digit Span test and Boston Naming test in the control group were higher than those in the case group, the Trail Making test time in the control group was shorter than that in the case group, and the differences were statistically significant ( P<0.05) . Conclusion:Acute paraquat poisoning can impair human cognitive ability to a certain extent.

Objective:Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity.Methods:Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue.Results:After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm 3 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion:Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.