Objective To appraise the diagnosis value of pleural membrane biopsy on pleural effussion.Meth- ods 218 patients with pleural effusion who were had pleual menbrane biopsy were retrospectively analyzed.Among them 165 cases were done only once,42 cases were done twice,11 cases were done three times.They were done 271 times of biopsies.Results The pleural membrane biopsy always masculine gender rate was 72.0%,1 pleural mem- brane biopsy succeeds the masculine gender rate was 59.6%,after the masculine gender rate which 2 pleural mem- branes biopsies succeeds was 69.7%.The illness complication formation rate was 8.7%.Conclusion Pleural mem- brane biopsy is a simply,safe and higher diagnosing-rate method.The different spot pleural membrane biopsy may cnhance diagnosis rate.

Objective:To develop non-immunized human phage display library.Methods:The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:The developed phage library is diversity and can be used for selecting humanized scFv.

Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation.
Cell Membrane ; China ; Cryopreservation ; Eosine Yellowish-(YS) ; Fluorescent Dyes ; Humans ; Infertility, Male ; diagnosis ; Male ; Osmotic Pressure ; Semen Analysis ; methods ; Sperm Head ; Sperm Motility ; Sperm Tail ; Spermatozoa ; Staining and Labeling ; Water

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Objective:Expression of protein TSLP and selection of full human anti-TSLP single chain Fv ( scFv).Methods:The cDNA of TSLP was amplified.The amplified target gene and the expression vector pET 101/D-TOPO were ligated , and then transformed into E.coli BL21.The protein was induced to expression by IPTG and purified and identified.The biotinylated TSLP protein was used as antigen to select of human TSLP scFv from a constructed human scFv library by phage display .Results: The size of amplified cDNA of TSLP was about 423 bp,and that of expressed protein was about 26 kD.Dot blot and Western blot results showed that the expressed protein was correct.The constructed human scFv library was enriched for three rounds using biotinylated TSLP as antigen by phage display.ELISA results showed that 35% scFvs had binding activity with TSLP.The scFvs with good binding activity were selected and identified by Western blot and sequencing.Conclusion: The full human scFvs against for TSLP were selected suc-cessfully.

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

OBJECTIVETo observe the therapeutic effect of moxibustion on AIDS patients of spleen-kidney yang-deficiency.

METHODSSixty-six cases of AIDS were divided into a treatment group and a control group, 33 cases in each group. All of the patients were treated with HAART, with moxibustion at Tianshu (ST 25), Shenque (CV 8), Zhongwan (CV 12), Guanyuan (CV 4) added in the treatment group for 3 months. Clinical symptoms and cell immunity were recorded before and after treatment.

RESULTSAfter treatment, the effective rate was 90.9% in the treatment group, which was better than 66.7% in the control group (P < 0.05). The improvement of the score for clinical symptoms in the treatment group was superior to that in the control group (P < 0.01). After treatment, the CD4 lymphocyte counts increased in the two groups, with no significant difference between the two groups (P > 0.05). Additionally, increase of total lymphocyte count in the treatment group was superior to that in the control group (P < 0.05).

CONCLUSIONMoxibustion can increase the therapeutic effect of HAART on AIDS patients and increase the total lymphocyte count.

Acquired Immunodeficiency Syndrome ; immunology ; therapy ; Adult ; Antiretroviral Therapy, Highly Active ; Female ; Humans ; Kidney ; physiopathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Moxibustion ; methods ; Spleen ; physiopathology ; Yang Deficiency ; immunology ; therapy

ObjectiveTo identify the early predictors of refractory epilepsy (RE). MethodsAll 173 epileptic patients with correct diagnosis and reasonable treatment were enrolled. The 106 patients were classified as drug non-responsive epilepsy (DNR-EP). The remaining 63 patients were classified as drugresponsive epilepsy (DR-EP). With multiple logistic regression, the clinical characteristics between the two groups were compared to identify the early predictors of RE. ResultsMultiple logistic regression analysis demonstrated that more than 10 seizures before treatment (OR =4. 46, 95％ CI 1.60-12. 40, P =0. 004),mental retardation at early time ( OR =19. 87, 95％ CI 3. 60-109. 78, P =0. 001 ) and abnormal electroencephalogram(EEG) with epileptiform wave after treatment ( OR =7.57, 95％ CI 2. 54-22. 56,P ＜0. 01 ) were independent predictors of RE.Response to initial therapy was a protective factor of RE (OR=0.05, 95％ CI 0.018-0. 139, P＜0.01). ConclusionPatients who have many seizures before treatment, mental retardation at the early time, epileptiform abnormality in EEG after treatment and who are resistant to initial therapy are likely to develop into refractory epilepsy.

HIV/STIs remain a major global public health problem. One of the global strategies for the prevention and control of HIV/STIs is to interrupt their transmission, which requires the public health methods based on scientific evidence and cost-effectiveness. The scale-up of male circumcision services in the priority countries of the HIV-prevention project in sub-Saharan Africa has been hampered by the scarcity of trained providers and relative technical difficulty of male circumcision techniques recommended by WHO and UNAIDS. Shang Ring is an innovative and disposable device for male circumcision, which has been safely used for over 600 000 males in China since 2006. Clinical studies of more than 3 000 cases of Shang Ring circumcision in China, Kenya, Zambia, and Uganda have demonstrated its safety, effectiveness, acceptability and ease of use. The most obvious advantages of Shang Ring include short procedure time (3-6 min), excellent postoperative cosmesis, low rate of complications, high acceptance by clients and providers, ease of use, and standardization for reliable performance. As an innovative technique, Shang Ring has a great potential for facilitating the safe and effective scale-up of circumcision services. This article comprehensively reviews the clinical studies of Shang Ring male circumcision in China and Africa.
Africa ; China ; Circumcision, Male ; instrumentation ; methods ; HIV Infections ; prevention & control ; Humans ; Male