Minimally Invasive Surgical Procedures ; Philosophy, Medical

Background The dysfunction of the blink reflex the eyelid-closure ability appears in the patients with facial paralysis,and its management is the implantation of mechanical-assisted eye-closure device in the upper eyelid.A novel device is palpebral spring implant.However,there is no similar study in China.Objective This study was to evaluate the clinical efficacy of palpebral spring placement for lagophthalmos caused by facial nerve palsy.Methods This clinical research complied with Helsinki declaration and the protocol was approved by Ethic Committee of Henan Eye Institute & Henan Eye Hospital.Written informed consent was obtained from each patient prior to the surgery.A retrospective serial case-observational study was performed.The medical records of 11 patients who underwent palpebral spring placement for hypophasis due to facial nerve palsy were reviewed at Henan Eye Hospital from August 2010 to November 2012.Palpebral spring placement was performed by the same surgeon to ensure a more even outcomes.Palpebral spring was made by nickel wire,with the diameter of 0.3 mm and implanted on tarsal plate in 11 eyes of 11 patients with symptomatic facial nerve palsy.The lower tip of Levine spring was encased into a small terylene bag and sutured to the anterior tarsal surface during the surgery.Preoperative and postoperative symptoms,upper eyelid margin to mid pupil distance (ULMD),degree of lagophthalmos and eyelid moving scope were examined and compared between before and after operation.The operating complication was followed-up for 8-38 months.Results The discomforted symptoms disappeared in all the operated eyes.The ULMD was (3.51±0.73) mm in preoperation and (3.20±0.86) mm in posteration,without significant difference between them (t=1.36,P=0.10).The degree of lagophthalmos was (5.94±1.57) mm and (1.06±0.98) mm in preoperation and postoperation respectively,showing a significant difference between them (t =9.42,P =0.00).The eyelid moving scope was (5.89±0.70) mm in postoperation,which was significantly increased in comparison with (0.11 ±0.33) mm of preoperation (t =22.97,P =0.00).The palpebral spring implant was regulated in 1 patient during the follow-up duration due to the trauma.No complication in other 10 patients appeared during the follow-up duration,such as implant exposure,metal fatigue or infection.Conclusions Palpebral spring placement is safe and effective for lagophthalmos in patients with facial nerve palsy.

Objective to investigate the correlation between glutathione S- transferase gene M1,t1 and P1(GStM1,GStt1 and GStP1)poly-morphism and sperm DNA fragmentation index(DFI)in male patients with idiopathic infertile. Methods the study included 246 male patients with idiopathic infertility. Polymerase chain reaction(PCR)and polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP) were used to identify the genotype of GStM1,GStt1 and GStP1,respectively. Sperm DFI was analyzed by flow cytometry with acridine orange staining. Results the sperm DFI was significantly higher in GStt1 gene null type group than in GStt1 gene wild type group(P < 0.01);and there were no significant differences in GStM1 and GStP1 gene mutation type groups when comparing with the wild type group(P > 0.05). Conclusion GStt1 gene deletion was positive associated with the sperm DFI in male idiopathic infertile patients.

Objective To detect the immunogenicity of VirB9,a protein of type Ⅳ secretion system of Brucella.Methods Full length VirB9 gene was cloned into plasmid pET32a and expressed in Escherichia (E.) coli BL21 (DE3).Expression of recombinant protein was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and the recombinant fusion protein was purified by affinity chromatography on Ni2+-conjugated chelateing sepharose.The purity of the purified protein was ascertained by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDSPAGE) and the concentration was measured by bicinchoninic acid (BCA) protein assay kit.Animal model was established by immunizing BAL B/c mice with live vaccine strain S19 of Brucella and the mice immunized with phosphate buffered saline (PBS) as control.The blood of immunized mice was acquired after 4 weeks.Antibody against VirB9 in S19 immunized mice was detected by Rose Bengal plate agglutination test and serum tube agglutination test; IgG antibody titers against VirB9 in immunized mice were determined by enzyme linked immunosorbent assay(ELISA).At the 35th day,the immunized mice and control mice were killed and spleens were collected.The splenocytes were harvested and stimulated with each of VirB9,concanvalin A(ConA) or medium in triplicate.Production of gamma interferon (IFN-γ) was determined by enzyme-linked immunospot assay (Elispot).Results The full length of VirB9 gene was cloned into pET32a.The recombinant VirB9 protein was expressed at 43 × 103 in relative molecular mass and the purity of the purified recombinant VirB9 protein was above 97％ in SDS-PAGE and the concentration was 1.6 g/L in BCA protein assay.The antibody of VirB9 was detected in all S19 immunized mice but not PBS immunized mice by Rose Bengal plate agglutination test.The antibody titer in all S19 immunized mice was ＞ 1 ∶ 800 or ＞ 1 ∶ 3200 by tube agglutination test and ELISA,respectively.Meanwhile,the protein stimulated stronger IFN-γresponse in immunized mice than that in the control mice(147 cells Vs 38 cells).Conclusion VirB9 can stimulate humoral and cellular immunity and it might be an appropriate target for developing subunit vaccine against Brucella.

This study was aimed to explore the role and mechanism of AML1a in abnormal hematopoiesis in mice. Plasmids pMSCV-FLAG-AML1a-IRES-YFP and pMSCV-IRES-YFP together with envelope-encoding plasmid pECO and packaging plasmid pGP were respectively transfected into 293T cells by using a method of calcium phosphate precipitation to produce retrovirus. Bone marrow mononuclear cells (BMMNC) from male C57BL/6J mice were transfected with the retroviral vector MSCV expressing FLAG-AML1a fusion protein and yellow fluorescent protein (YFP). The cells were cultured in M3434 semi-solid medium for colony formation assay and in M5300 fluid medium containing murine IL-3 (mIL-3), IL-6 (mIL-6) and SCF (mSCF) for long-term culture. The results showed that transfection of AML1a into BMMNC enhanced colony formation, colony size of the AML1a group was significantly larger than that of the control group, and the colonies were mainly composed of CFU-E and CFU-GEMM. In the long-term culture, AML1a-transfected BMMNC showed differentiation block, while the control cells were in a more mature stage. It is concluded that AML1a may block the normal hematopoiesis at the stage of primitive progenitors. At the same time, AML1a also enhances the proliferation activity of primitive progenitor cells.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Proliferation ; Colony-Forming Units Assay ; Core Binding Factor Alpha 2 Subunit ; genetics ; Genetic Vectors ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Retroviridae ; genetics ; Transfection

The aim of this study was to investigate the feasibility of using fetal short tandem repeat (STR) loci in maternal plasma as gender-independent fetal DNA marker. DNA from maternal plasma sample was extracted using QIAamp DNA Kit. AmpF1 STR profiler box was used to amplify 9 different polymorphic short tandem repeat (STR) loci (D3S1358, VWA, FGA, D5S818, D13S317, D7S820, D8S1179, D21S11, D18S51), the multiplex fluorescent PCR was used to amplify the STR alleles of fetal DNA in 36 pregnant plasma samples of pregnant women at different pregnancy. Their husbands' DNA isolated from whole blood samples were amplified at the same time. The PCR products were electrophoresis by ABI Prism 377 sequencer, the results of electrophoresis were analysed by Genscan. The presence of fetal DNA in maternal plasma by Paternally inherited fetal alleles were detected. The results showed that paternally inherited fetal alleles were detected in 4 cases in early pregnancy (4/6), 19 cases in middle pregnancy (19/20) and 9 cases in late pregnancy (9/10) respectively, the paternally inherited fetal alleles in 4 of 36 cases could not be detected. It is concluded that fluorescent multiplex PCR can be used for amplification of male and female fetal STRs in maternal plasma to obtain genetic information, which may have implication for non-invasive prenatal diagnosis of certain hereditary diseases independent of the fetal sex.
DNA ; analysis ; Female ; Fetus ; Genetic Markers ; Genotype ; Humans ; Male ; Microsatellite Repeats ; Plasma ; chemistry ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; Sex Characteristics

AIMTo explore the changes of mRNA and protein expressions of heart-type fatty acid binding protein (H-FABP) in rat ischemic myocardium at different intervals ischemia.

METHODS60 SD male rats weighing 250-350 g, were randomly divided into one sham-operated group and five study groups (group A1, A2, A3, A4, A5, the left coronary artery of rats has been ligated for 1 h, 2 h, 4 h, 6 h, 12 h respectively). Myocardil samples from infarct zone, ischemic and non-ischemic zone, were obtained for histology examination, and the mRNA for H-FABP in ischemic myocardial tissue were determined by RT-PCR. Serum free fatty acid(FFA) was determined by colorimetric method.

RESULTSCompared to sham hearts, H-FABP mRNA expression were significantly decreased in ischemia zone of AMI rat hearts (P < 0.05), especially in rats underwent 4 h ischemia and 6 h ischemia (P < 0.01). Serum FFA were significantly increased in AMI rats relative to sham rats (P < 0.05).

CONCLUSIONSignificant down-regulated heart-type fatty acid binding protein after myocardial ischemia might play an important role in myocardial injury and energy metabolism disorder.

Animals ; Down-Regulation ; Fatty Acid-Binding Proteins ; genetics ; metabolism ; Male ; Myocardial Infarction ; metabolism ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.

METHODSGenomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.

RESULTSThe sequencing results showed HLA-DRB1 alleles of the proband as DRB1*090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1*120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.

CONCLUSIONThis allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; ethnology ; DNA ; analysis ; Ethnic Groups ; Female ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis

OBJECTIVETo explore therapeutic effects of bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle,and analyze manipulative reduction mechanism.

METHODSFrom 2005 to 2008, 95 patients with over degree II supination-eversion fractures of ankle were treated respectively by manipulation and operation. There were 43 cases [11 males and 32 females with an average age of (44.95 +/- 12.65) years] in manipulation group, and 2 cases were degree II, 11 cases were degree III, and 30 cases were degree IV. There were 52 cases [21 males and 31 females with an average age of (39.96 +/- 13.28) years] in operative group,and 6 cases were degree II, 18 cases were degree III, and 28 cases were degree IV. Bone setting manipulation and hard splint external fixation were applied to manipulative group. Operative reduction internal fixation was performed in operative group. X-ray was used to evaluate reduction of fracture before and after treatment, 2 months after treatment. Ankle joint function was evaluated according to Olerud-Molander scoring system after 6 months treatment.

RESULTSAll patients were followed up with good reduction. Three cases occurred wound complication in operative group, but not in manipulative group. In manipulation group, 19 cases got excellent results, 20 cases good and 4 cases fair; while in operative group, 30 cases got excellent results, 20 cases good and 2 cases poor. There were no significant differences in fracture reduction and ankle joint function recovery between two groups (P > 0.05). Efficacy of operative treatment was better than that of manipulative treatment at degree IV fracture (P < 0.05).

CONCLUSIONBone setting manipulation is a good method for treating supination-eversion ankle joint fractures, which has advantages of simple and safe operation, reliable efficacy. For ankle join fracture at degree IV, manipulative reduction should be adopted earlier, and operative treatment also necessary

Adult ; Ankle Injuries ; physiopathology ; therapy ; Ankle Joint ; physiology ; Case-Control Studies ; Female ; Fractures, Bone ; physiopathology ; therapy ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Supination

China ; Humans ; Mycobacterium tuberculosis ; isolation & purification ; Quality Control ; Sputum ; microbiology ; Tuberculosis, Pulmonary ; microbiology