The progress of investigation on medicinal values of hipocampus, indluding its chemical components, pharrnacologic action, clinical application and so on were summarized in this paper.

Objective To establish a multiplex real-time quantitative polymerase chain reaction (MRQPCR) assay for fast and simultaneous detection of Escherichia coli (E.coli) and Candida albicans (C.albicans) genes in human whole blood,in order to facilitate differentiation of the types of microorganism and evaluation of the severity of bacterial or fungi translocation due to impaired gut barrier,hence providing help to select specific antimicrobial agents.Methods The β-D-galactosidase gene of E.coli and ITS2 gene of C.albicans were selected as the target genes for designing primers and probes.E.coli and C.albicans genomes were extracted with QIAamp(R) DNA Blood Mini Kit,and the 25 μl TaqMan MRQ-PCR amplification reaction system was established.18 simulated human whole blood samples and 10 whole blood samples from febrile surgical patients were detected for E.coli and C.albicans genes using MRQ-PCR.Results The specificity of the primers and probes were excellent.The correlation coefficients of the standard curves of E.coli and C.albicans were 0.994-0.999 and 0.994-0.998,respectively;and the efficiency of amplification were 0.894-1.022 and 0.905-1.028,respectively.In the standard samples,the lowest detection limits of E.coli and C.albicans were 13.9 copies/μl and 0.8 cfu/μl,respectively;the sensitivity was 100％ and 99.69％,the specificity was 100％ and 94.73％,respectively;the average recovery rates were (101.89 ± 5.69)％ and (103.74 ± 4.64)％ respectively;the intra-batch coefficients of variance (CV) in detecting the genes were (13.14 ± 10.27)％ and (19.18 ± 8.54)％,respectively,and the inter-batch CV were (14.35 ± 9.34)％ and (18.31 ± 10.25) ％,respectively.In human whole blood,the lowest detection limits of E.coli and C.albicans were 12 455.2 copies/ml and 800.3 cfu/ml,respectively;the average recovery rates were (111.60 ± 11.06) ％ and (99.96 ± 6.16) ％,respectively;the intra-batch CV in detecting the genes were (11.02 ± 5.65) ％ and (8.14 ± 7.29)％,respectively,and the average inter-batch CV were (12.88 ± 7.59)％ and (18.62 ± 9.14)％.Conclusions MRQ-PCR is a rapid,sensitive,specific,accurate,and reproducible method for simultaneous detection of E.coli and C.albicans genes in human whole blood,with sample-,cost-,and time-saving advantages.It is a promising technique for rapid differentiation between fungi and bacteria,which could help targeted administration and evaluation of antimicrobial agents,and help to assess the consequence of gut barrier damage and the efficacy of treatment.

Objective To establish a rapid real-time quantitative polymerase chain reaction (RQ-PCR)assay in quantifying and detecting Staphylococcus aureus DNA from human venous blood samples,so as to quantificationally evaluate the systemic infection caused or deteriorated by intestinal bacteria translocation.Methods Totally 26 clinical blood samples and 15 simulation blood samples were detected.The primers and TaqMan probe were designed targeting the highly conserved house-keeping femA gene of Staphylococcus aureus,and a 20 μl RQ-PCR amplification reaction system was established.The standard curve was built based on the recombinant plasmid DNA containing the amplicon of the target gene,and genomic DNA was extracted using QIAamp DNA Blood Mini Kit.Results The specificity of primers and probe was excellent,the detecting limit was 100 copies/μl (103 CFU/ml),the sensitivity was 99.7％,and the specificity was 94.6％.The correlation coefficient of the standard curve was between 0.9918 and 0.9997.For samples with different Staphylococcus anreus concentrations,the average accuracy of the RQ-PCR assay was (96.25 ± 2.26) ％ ; the intra-and interassay coefficients of variation were (8.06 ±0.07)％ and (10.01 ±4.40)％,respectively.The average recovery rate of Staphylococcus aureus DNA in blood samples was (111.72 ± 20.72) ％.In clinical blood samples,the positive rate of Staphylococcus aureus DNA was 15.4％ (4/26),while the blood culture of these samples all produced negative result for Staphylococcus aureus.Conclusion RQ-PCR assays is a rapid,sensitive,and specific method with good repetitiveness and can be used in the quantitative detection of Staphylococcus aureus in whole blood samples.

Aim To obtain the encoding gene sequences of TEM-type ?-lactamases produced by 4-strain Klebsiella pneumoniae in Zhejian g Province, identify their genotypes and study some properties of these TEM-typ e ?-lactamases.Methods The encoding genes of TEM-type ?-la ctamases produced by 4 isolates were amplified by PCR. The purified PCR products were ligated with pGEM-T easy vectors, expressed in E. coli DH 5?, and sequenced by Sanger's dideoxy chain termin ation composition method. Compared with anino acid sequences in the GenBank,TEM -types of the ?-lactamases was determined. The genes of TEM ?-lactamases were ligated with pET-28 c vector to express recombinant proteins in E. coli DH 5?. Plasmids were extracted from the p ronucleus expression strains and PCR was performed to determine whether the pron ucleus expression was successful or not. Their phenotypes were determined by ESB Ls phenotype affirmative test. The isoelectric points (pIs) of the recombinant p roteins were determined by isoelectric focus. Conjugation test was performed to determine whether their genes existed in plasmid or chromosome. Results The encoding genes of ?-lactamases were determined as TEM by PCR. It s PCR product had 1 009 nucleotides. The pI of the novel TEM ?-lactamase was 5.4. The enzyme was determined as non-ESBLs by ESBLs phenotype affirmative tes t.Transconjugants were successfully selected from the paternal producers in conj ugation tests. The TEM-type ?-lactamase produced by 4 strains was determined as TEM-105(AF516720) by GenBank. Conclusion The ?-lactamase produced by 4-strain K. pneumoniae from 4 patients in Zhejiang Province was TEM-105. It was the first report of TEM-105 type ?-lactamase produced by 4-st ain K. pneumoniae from China in the world.

Objective To establish a real-time quantitative polymerase chain reaction (RQ-PCR) assay for fast detection of Aspergillus fumigatus genome in human whole blood samples and explore its clinical application.Methods The primers and the TaqMan-probe were designed on the basis of the multi-copy ITS1-5. 8S region of the rDNA of Aspergillus fumigatus. The Aspergillus fumigatus genomic DNA were extracted with QIAamp(R) DNA Blood Mini Kit.A 20 μl RQ-PCR amplification system was established, and the simulated blood samples containing various given load of Aspergillus fumigatus genome and the 66 whole blood samples of the surgical febrile patients were examined. Results The detection limit of the RQ-PCR instrument is 10-1 genomes/μl DNA sample,namely 78 CFU/ml whole blood. The specificity and the sensitivity were 94. 25% and 99. 04% respectively; and the positive predictive value and negative predictive value were 97. 63% and 97. 62% respectively. The average relative error of the quantitative results was (3. 67 ±13. 19)%, and the intra- and the inter-assay average coefficients of variation were (12.38 ± 1. 53)% and (16. 27 ±2. 72)% , respectively. The average recovery rate of Aspergillus fumigatus genomic DNA in human whole blood samples was (107. 81 ±25. 92)% , and the average coefficient of variation of the average recovery rate was (26. 24 ± 5.62) % . No Aspergillus fumigatus genomic DNA was detected among the 66 blood samples of the surgical febrile patients. Conclusions The RQ-PCR assay for fast quantitative detection of Aspergillus fumigatus genome in human whole blood samples is of high sensitivity, specificity,accuracy and precision. The Aspergillus fumigatus genome was not detected in this group of surgical febrile patients.

Objective To investigate the effect of chemotherapy drugs on the expression of carci-noembryonic antigen (CEA) in the gastric tissue with precancerous lesions in ectopie cancer. Methods There were 45 cases of cancer patients (precancerous lesions group), the pathological biopsy showed that there were atypical hyperplasia or intestinal metaplasia by gastroscope before chemotherapy. Gastruscope was done before chemotherapy and after six cycles of chemotherapy. Gastric tissue was taken respectively in the same site. The expression of CEA was measured in the gastric tissue. Normal gastric tissue taken from 10 cases of cancer patients was served as control. Compared respectively the expression of CEA in the gastric tissue in control group and precancerous lesions group, in precancerous lesions group between before and after treatment. Results CEA expression in the gastric tissue was (27.76±9.67), (3.32±0.60)μg/L in precancerous lesions group and control group respectively, there was significant difference between two groups(P<0.05). CEA expression in the gastric tissue was (27.76±9.67), (26.60±10.80)μg/L before and after treatment in precancerous lesions group respectively, P<0.05. CEA expression in the gastric tissue before treatment was (23.11±4.11), (17.10±1.66)μg/L, after treatment was (21.11±5.66), (15.10±3.31)μg/L in the mild to moderate atypical hyperplasia, mild to moderate intestinal metaplasia respectively, there was significant difference between before and after treatment in the mild to moderate precancerous lesions. There was no significant difference between before and after treatment in the severe precancerous lesions. Conclusions Chemotherapy drugs can significantly reduce the expression of CEA in the gastric tis-sue in the mild to moderate precancerous lesions. The results suggests that mild to moderate precancerous lesions can be reversed.

Objective To establish the real-time quantitative PCR (RQ-PCR) assay for detecting Candida albicans (C.albicans) in whole blood and its clinical application in the febrile surgical patients who may develop gut barrier damage and gut microorganism translocation.Methods The NAG1 gene,which is a single copy in C.albicans genome,was selected as the target gene for designing the primers and probe.The plasmid was fabricated and produced as standard samples.C.albicans genomes were extracted with QIAamp(R) DNA Blood Mini Kit,and the total 20 μl TaqMan RQ-PCR amplification reaction system was established.The 74 venous blood samples from the surgical febrile patients were detected for C.albicans load.Results The specificities of the primers and probe were excellent,the correlation coefficients of the standard curves were between 0.9918 and 0.9985,and the efficiency of amplification was 0.88-1.027 for the samples above the lowest detection limit (100 copies/μl examine fluid,or nearly 1.1 × 103 cfu/ml whole blood).The average accuracy of the RQ-PCR equipment was (99.64±2.08) %,the sensitivity was 97.46%,the specificity was 100%,and the average coefficients of variation (CV) of the intra-and inter-assay were (14.76±2.64)% and (17.85±3.53)%,respectively.The average recovery rate of C.albicans DNA in whole blood samples was (88.60±5.73) %,and the average CV of recovery rate was (11.70 ±5.36) %.The number of copies of C.albicans genes per unit blood was not significantly different among the same original blood samples stored separately under-20℃ for 3 or 6 months when compared with its freshly collected blood (P = 0.267).In the 74 whole blood samples obtained from the febrile surgical patients,the positive rate of C.albicans genes was 2.7% and the highest load was 4.42×103 cfu/ml.Conclusions RQ-PCR is a rapid,sensitive,highly specific,and reproducible method in detecting C.albicans NAG1 gene.Clinically it can be used to quantitatively evaluate the numbers of C.albicans in the whole blood.A small percentage of the febrile surgical patients may develop blood infection of C.albicans.

Objective:To investigate the epidemiological and clinical features of patients with hemorrhagic fever with renal syndrome (HFRS) from 2017 to 2019.Methods:Seventy-five patients with HFRS from the Department of Infectious Diseases, The First Affiliated Hospital of Anhui Medical University during January 1, 2017 to December 31, 2019 were included. The data of epidemiology, clinical symptoms, blood routine, urine routine, serum creatinine, liver function and other laboratory examination indexes were retrospectively analyzed. The measurement data with skewness distribution were expressed by M( QR) and compared by nonparametric test. Multivariate logistic regression analysis was used to analyze disease-related risk factors. Results:The 75 patients were mainly located in the western and northern regions of Anhui Province. A total of 37 cases (49.3%) were infected during November, December and January next year. Fifty-four (72.0%) patients were farmers and 10(13.3%) patients had a clear history of rodent contact. Only 19(25.3%) patients had typical clinical manifestations of "three red and three pain" . Fifty-eight (77.3%) patients had elevated white blood cell count, 67(89.3%) patients had decreased platelet count, 55(73.3%) patients had urinary protein + + + , 65(86.7%) patients had abnormal urinary occult blood, and 67(89.3%) patients had elevated serum creatinine. The serum creatinine and potassium levels in 31 severe and critical patients were 495(301) μmol/L and 4.14(0.77) mmol/L, respectively, which were both higher than those in 44 mild and moderate patients (235(289) μmol/L and 3.65(1.02) mmol/L, respectively). The differences were both statistically significant ( Z=-3.187 and -2.796, respectively, both P<0.01). Multivariate logistic regression analysis showed that serum creatinine (odds ratio ( OR)=1.005, 95% confidence interval ( CI)1.002-1.008) and serum potassium ( OR=2.632, 95% CI 1.098-6.313) were independent risk factors for disease severity. All patients received comprehensive medical treatment, and 27 patients received renal replacement therapy. Sixty-eight patients had good prognosis and four patients died. Conclusions:HFRS is still common in the rural area in winter and spring. Patients with atypical clinical manifestations and severe and critical patients should be intensively monitored.

Background The operation mode of automobile manufacturing industry (AMI) makes workers have different degrees of occupational stress and burnout, which may lead to negative emotions and depressive symptoms. Objective To study the relationship between occupational stress, job burnout, and depressive symptoms in AMI workers. Methods In this study, 1300 workers from a Guangzhou AMI company were selected as subjects by cluster random sampling method. Occupational stress, job burnout, and depressive symptoms of the workers were assessed by using the Effort-Reward Imbalance (ERI) questionnaire, the Maslach Burnout Inventory general survey questionnaire, and the Patient Health Questionnaire-9, respectively. Hierarchical regression was used to analyze the effects of occupational stress and job burnout on depressive symptoms in AMI workers. Mediating effect model was used to analyze the mediating effect of job burnout on the relationship between occupational stress and depressive symptoms. Results There were 1300 questionnaires distributed, 1228 valid questionnaires collected, with a 94.5% recovery rate. The ERI ratio of 1228 AMI workers was 1.06±0.72, and the positive rate of occupational stress was 37.3% (458/1228). The score of job burnout was 2.18±1.37, and the positive rate of job burnout was 62.6% (769/1228). The score of depressive symptoms was 10.27±6.42, and the positive rate of depressive symptoms was 47.1% (578/1228). The dimensional scores of effort and over-commitment in occupational stress as well as emotional exhaustion and depersonalization in job burnout of AMI workers were positively correlated with the depressive symptom scores (r_s=0.415, 0.571, 0.573, 0.593, P＜0.05). The dimensional scores of reward and personal achievement were negatively correlated (r_s=−0.454, −0.339, P＜0.05). The percentages of variance in depressive symptoms score explained by occupational stress and job burnout were 26.7% and 16.6%, respectively. Job burnout had a partial mediating effect between the three dimensions of occupational stress and depressive symptoms, and the mediating effect values were −0.2832 (95%CI: −0.3250– −0.2434), 0.3553 (95%CI: 0.3071–0.4041), and 0.4193 (95%CI: 0.3681–0.4725), respectively. Conclusion AMI workers' occupational stress affects job burnout, but also indirectly affects depressive symptoms. Job burnout partially mediates the association between occupational stress and depressive symptoms. Reducing occupational stress and burnout levels of AMI workers may alleviate depressive symptoms.

ObjectiveTo explore the effect of the calcium phosphate cement (CPC) /calcium polyphosphate fiber (CPPF) composites mixed with different proportion of minimal morselized bone on repair of bone defect in vivo. MethodsA total of 36 New Zealand white rabbits were completely randomly designed into A, B, C, D groups and their bilateral radial bone defect model was prepared. The minimal morselized bone (300-500 μm in diameter) was made from the iliac of those rats. The CPPF and CPC were evenly mixed into CPC/CPPF composites which were divided into four groups in accordance with the CPPF weight O, 10％, 30％ and 50％ in CPC/CPPF composite. The CPC/CPPF composites of the four groups was mixed with the minimal morselized bone with ratio of 6:4 and then the mixture was implanted the bone defect of the rabbits in four groups. The gross, X-ray and histological observations were done at four and eight weeks. The biomechanical test was performed at eight weeks. Results When CPPF occupies 30％ of the CPC/CPPF composite, the maximum compressive load and bending loads were better than those in the other groups ( P ＜ 0.05 ), when the histological observation showed the most tight link between the artificial composite and the bone interface and the closest similarity between material degradation rate and the ossification rate, with the best osteogenesis and the optimal ratio.ConclusionThe repair of bone defect can attain the optimal outcome through adding a certain ratio of minimal morselized bone into the CPC/CPPF to adjust the degradation rate of composites.