1.Establishment of ELISA method of detecting human cytomegalovirus by use of recombinant antigen pp65 and the comparative analysis with two other methods
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To establish a new ELISA method by use of shorted recombinant antigen pp65 for detection of IgM antibodies to human cytomegalovirus (HCMV).Method According to HCMV pp65 sequence of nucleotide and amino acid, the shorted pp65 was decided by computer soft ware of protein and attained through gene engineering technique. The by computer soft ware of protein and attained through gene engineering technique. The recombinant enzyme- linked immunosorbent assay (REC-ELISA ) method was developed using pp65 recombinant protein as antigen and was applied to detecting anti-HCMV-IgM in sera. It was compared to ELISA by use of whole virus as antigen and Biocheck CMV IgM enzyme immunoassay test kit.Results To detect HCMV IgM by REC-ELISA, the best quantity of pp65 was 3.5?g percent cavity, the best dilution of HRP~*anti-HCMV IgM was 1∶[KG-*2]800 and that of serum was 1∶[KG-*2]100. The positive coefficient of variation (CV) was 9.5% in stability assay. The average CV of Inter- assay and Intra-assay was 4.5% and 9.6% respectively. The positive value of REC- ELISA was 44%, that of ELISA using whole virus was 50% and BioCheck was 45%. REC-ELISA using most suitable condition was concordant with BioCheck, which was 97.0%. Its youden′s index was 92.8% and its specificity (98.2%) was higher than that of ELISA using whole virus as antigen (90.9%), too.Conclusion REC-RLISA had good sensitivity, specificity and reproducibility. The method was easy and quick. Recombinant protein was harmless and easy gained compared with the whole virus. It can be large-scale production and standardization. Therefore, the application value and potential of REC-ELISA was very large.
2.The liver and spleen interventional therapy for primary carcinoma of the liver with hyperspleenism
Chinese Journal of Primary Medicine and Pharmacy 2010;17(8):1033-1034
Objective To study the value of the liver and spleen interventional therapy in primary carcinoma of the liver with hyperspleenism. Methods The treatment was given to 25 patients. Using the method of Seldinger,the 5FRH into artery hepatica propria to infuse 2/3 of the overall dose of chemotherapy medicine and embolism was inserted into the liver segment. Then 1/3 of it was infused into artery lienalis and gelatin sponge was infused into artery lienalis Rr. lienales. Combined regimens were used in chemotherapy: 5-Fu 750mg, EADM 40mg, MMC 10mmg,and 10 of the 25 added DDP 40mg. LUF and gelatin sponge were used for embolisation. Results Of the 25 patients,the WBC and PLT increased in 25 patients 4 weeks after the operation. The difference was significant. The survival rate at 6 and 12 months was 100% and 60% respectively,with 1 case of CR,8 cases of PR and 6 cases of SD. Patients had fever after the treatment but few with abnormal liver function tests. Conclusion Liver and spleen interventional therapy may relieve hyperspleenism and suggests that the treatment of primary carcinoma of the liver is effective.
3.The effect of the human cytomegalovirus pp65 IgG-avidity index for the human cytomegalovirus primary infection in mice model
Chinese Journal of Laboratory Medicine 2008;31(5):557-561
Objective To investigate the significance of human cyomeg Movirus(HCMV)pp65 IgG antibody avidity index(AI)for the clinical diagnosis of HCMV primary infection through the experimental model of HCMV primaly infection in BALB/c mice.Methods 6~8 weeks,female,specific-pathogen-free BALB/c mice were divided into 5 groups.6 mice in each group. And he mice were injected with 2×106 PFU/m1,2×105 PFU/mi,2×104 PFU/ml,2×103 PFU/ml and 2×102 PFU/ml of HCMV intraperitoneally respectively. Another 6 mice were injected intraperitoneally with the maximum dose of HCMV kept at 56℃ for 30 min as inactivated virus group.And HF negative control group was established at same time. All the mice ere sacrificed to obtain brain and lung tissues for the following experiments after 1 montll.(1)Tissue samples obtained from mice were inoculated in human embryo fibroblasts(HF)monolayers after routine treatment for virus isolation.HCMV specific eytopathie effect(CPE) was observed bv inverted phase-contrast microscopy.HCMV UL83 DNA in the ultures as tested by PCR and pp65 antigen was detected by indirect immunofluorescence.(2)Extracted mRNA from tissue samples and HCMV pp67 mRNA were analyzed by reverse transcriptase PCR(RT-PcR).(3)Immunoglobulin M(IgM)antibody and immunoglobulin G(1gG)antibody avidity Was investigated for their usefulness in distinguishing primary genital HCMV infections rom nonprimary infections with ELISA kit using truncated pp65 protein.ResultsHCMV can be isolated in the tissues from the mice injected with 2×106 PFU/ml and 2×105 PFU/m1.RT- PCR and ELISA showed positive results in the same groups.The infective rates were 100%.The analysis of the low doses groups,inactivated group and HF negative ontrol group all showed negative results.Conclusions BALB/c mice can be infected with HCMV and appeared as primary infection after1 month.Determination of HCMV pp65 IgM and HCMV p065 IgG-AI by ELISA incorporated with virus isolation and RT-PCR are helpful for distinguishing primary infections from nonpfimary infections.The detection of HCMV p65 IgM and HCMV pp65 IgG-AI by ELISA utilizing recombinant protein pp5 as antigens can be used for preliminary screening.
5.Character of early tinnitus about sudden deafness.
Rongguo WANG ; Mingli GUO ; Yingzhang MIAO
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(10):909-911
OBJECTIVE:
To study the clinical characteristics of early tinnitus in patients with sudden deafness.
METHOD:
Forty-seven patients with sudden deafness and early tinnitus recieved subjective review and filled out classified questionnaire of tinnitus. By pure tone audiometry and tinnitus matching test, we obtain the distribution, quantity and matching loudness of tinnitus patients.
RESULT:
(1) Objective examination: the most common frequencies of tinnitus were 8000 Hz and 4000 Hz. The most common numbers were two frequencies. The matching loudness was (16.63 ± 8.37) dB. (2) Subjective examination: the tinnitus was monotone (74.0%), sustained (89.0%), third level loudness (45.0%), third level annoying (60.0%), high frequency (83.0%), sound of cicadas (66.7%). The most influencing factors were hearing (28.2%) and mood (29.5%). (3) Relationship between tinnitus and pure tone hearing threshold curve: there was a significant correlation between the frequency of pure tone audiometry and the matching loudness of tinnitus (r = -0. 370, P < 0.01). There was no significant correlation between the frequency of pure tone tinnitus and the matching loudness of tinnitus (r = -0.083, P > 0.05).
CONCLUSION
(1) The results of subjective examination were consistent with those of objective examination. (2) The tinnitus should be considered in the early treatment of sudden deafness.
Audiometry, Pure-Tone
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Hearing Loss, Sudden
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pathology
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Humans
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Tinnitus
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pathology
6.The three-dimensional measurements and analysis of anatomic artificial teeth cusp inclination
Mingli LIU ; Yong WANG ; Xiaodong CHEN
Journal of Practical Stomatology 2015;(6):789-792
Objective:To survey the continuous change features of cusp angle in anatomic artificial teeth,and to investigate their effects on occlusal balance.Methods:28 artificial teeth of a complete denture were measured by optical three-dimensional(3D)scan-ner and the corresponding digital models were constructed using the reverse engineering software Surfacer 1 0.5.The working side cusp slopes of lateroprotrusive movement including the maxillary buccal cusp distal slopes and the mandibular buccal cusp mesial slopes were studied.The straight lines from bottom to top of cusp were constructed and the corresponding projected curves in occlusal surface were generated.The angulation values of straight line and the continuous change values of projected curve were calculated.Results:The angulation values of artificial teeth cusp inclination including straight line and corresponding projected curve were obtained.The maxillary working cusp inclination varied from 1 3.7°to 25.6°,and the mandibular working cusp inclination varied from 1 1 .3°to 37.7°.Conclusion:The relatively large differences of the cusp inclination among traditional anatomic artificial teeth and the convex form of the cusp are disadvantage factors to set up stable centric stop and unhindered chew course.
7.Research on nucleotide sequence of a newly emerged pandemic norovirus GⅡ.4 genotype
Mingli ZHOU ; Ailing CAI ; Xuefeng WANG
International Journal of Laboratory Medicine 2017;38(2):231-232
Objective Analysis of the complete genome sequence about the newly emerged pandemic norovirus GⅡ.4 genotype, to understand its variation characteristics.Methods 264 patients were collected with diarrhea.The RNA was extracted from 264 fe-cal specimens and cDNA was synthetized.The positive samples were amplified by PCR,the amplified fragments were sequenced. The complete genome sequences of the norovirus was sequenced and analyzed.Results A new norovirus variant strain of Jingzhou GⅡ.4,a pleiston of Sydney GⅡ.4 was isolated.A large variation was found in the new variant subtype,which was mutated inclu-ding in the hypervariable P2 domain of the major capsid protein VP1.Conclusion The result demonstrates the variant strain of Sydney GⅡ.4 was spread to China.VP1 of norovirus GⅡ.4 is evolving rapidly.The spread and evolution situation of the norovirus GⅡ.4 need to be closely monitored in China for the development of effective vaccines and therapeutic monoclonal antibodies.
8.Application of automatic photography in Schistosoma japonicum miracidi-um hatching experiments
Mingli ZHOU ; Ailing CAI ; Xuefeng WANG
Chinese Journal of Schistosomiasis Control 2016;28(3):241-243
Objective To explore the value of automatic photography in the observation of results of Schistosoma japoni?cum miracidium hatching experiments. Methods Some fresh S. japonicum eggs were added into cow feces,and the samples of feces were divided into a low infested experimental group and a high infested group(40 samples each group). In addition,there was a negative control group with 40 samples of cow feces without S. japonicum eggs. The conventional nylon bag S. japonicum miracidium hatching experiments were performed. The process was observed with the method of flashlight and magnifying glass combined with automatic video(automatic photography method),and,at the same time,with the naked eye observation meth?od. The results were compared. Results In the low infested group,the miracidium positive detection rates were 57.5% and 85.0%by the naked eye observation method and automatic photography method,respectively(χ2=11.723,P<0.05). In the high infested group,the positive detection rates were 97.5%and 100%by the naked eye observation method and automatic pho?tography method,respectively(χ2= 1.253,P > 0.05). In the two infested groups,the average positive detection rates were 77.5% and 92.5% by the naked eye observation method and automatic photography method,respectively(χ2 = 6.894,P <0.05). Conclusion The automatic photography can effectively improve the positive detection rate in the S. japonicum miracidi?um hatching experiments.
9.Detection of Mycoplasma Caused Urogenital Tract Infection and Their Antibiotic Susceptibility
Mingli ZHOU ; Ailing CAI ; Xuefeng WANG
Chinese Journal of Nosocomiology 2006;0(05):-
OBJECTIVE To know about the urogentital tract infection caused by Ureaplasma urealyticum(Uu) and Mycoplasma hominis(Mh) and distribution and antibiotic susceptibility.METHODS Uu and Mh were isolated and their antibiotic susceptibility was detected by ICS kit from Lizhu company in Zhuhai.RESULTS Among the 246 samples,126 were positive(51.22%) of which Uu were 114,Mh 2,and both Uu and Mh 10;females were more apt to be infected(P
10.Change of opioid peptide content in plasma and CSF of patients with cerebral hemorrhage and interference effect of Naloxone
Mei HE ; Xuelian GUAN ; Mingli WANG
Journal of Clinical Neurology 1993;0(03):-
Objective To explore the change of opioid peptide content such as ?-endorphin(?-EP), dynorphinA_ 1-13(DynA_ 1-13) in plasma and cerebrospinal fluid(CSF) of patients with cerebral hemorrhage, and the interference effect of Naloxone on them.Methods 60 patients with cerebral hemorrhage were randomly divided into Naloxone-treated group and control group. Conventional treatment was used in both groups, then Naloxone-treated group intravenous drip of Naloxone, each 3.0 mg, once a day, for 14 d. The concentrations of ?-EP, DynA_ 1-13 of the patients were measured with radioimmunoassay (RIA) during the acute stage before treatment and at 7th, 14th day after treatment compared with those of normal group (the normal people and the patients were treated by operations). Changes of ?-EP and DynA_ 1-13 in bleeding part and amount of bleeding were analyzed. The scores of GCS and NDS before and after treatment were examined at the same time.Results (1) The content of ?-EP, DynA_ 1-13 in plasma and CSF of patients with cerebral hemorrhage were obviously higher since acute stage than those of the normal group(all P