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Objective To investigate the regulation effects of Ngn2 gene transfection on retinal neuron differnetion in three-dimentional optic vesicle (OV) of mice.Methods OV was cultured in vitro using mouse induced pluripotent stem cells (iPSC) under specific conditions.During OV culture,it was transfected multiple times by lentivirus-mediated Ngn2 gene and then it was induced after maturation.The cells were specificly differentiated toward retinal nerve cells in OV.Using the green fluorescent protein (EGFP) gene as control,the differentiation of retinal nerve cells in OV was detected by immunohistochemistry.Reverse transcription PCR and Western blot were used to quantitatively detect the expressions of retinal neuron-specific proteins Pax6,Islet1 and Brn3b.Results The mouse iPS-derived OV was successfully cultured.The number of neural cells in the OV transfected with the Ngn2 gene was increased by the lentiviral-mediated lentivirus.The expressions of PAX6,Islet1 and Brn3b in the Ngn2 transfection group were significantly higher at the gene and protein levels than those in the control group,with significant differences between the two groups (P＜0.05).Conclusions The Ngn2 gene can effectively increase the number of retinal neuron differentiation in OV and make in vitro cultured OV more mature and form a more perfect retinal cell neural circuit.
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Objective? To? analyze? the? ineffective? triggering? caused? by? nebulization? in? the? way? of?respiratory?mechanics.? Methods? A?test-lung?and?an?oxygen-driven?jet?nebulizer?were?connected?to?the?circuit?in?a?PB840?ventilator.?The?test-lung?was?pulled?outwards?in?manual?way?till?an?inspiration?was?effectively?triggered?separately?in?different?flow-trigger?modes?[flow-trigger?sensitivity?(VTrig)?3?L/min?and?5?L/min]?and?pressure-trigger?modes?[pressure-trigger?sensitivity?(PTrig)?2?cmH2O?and?4?cmH2O,?1?cmH2O?=?0.098?kPa]?with?the?nebulizer?being?closed?and?opened?in?turn.?The?corresponding?relationship?and?characteristics?between?the?flow?and?pressure?in?the?circuit?under?different?triggering?conditions?were?observed?by?adjusting?the?curve?amplitude?in?the?screen.?The?minimum?alveolar?pressure?(Pa)?which?could?cause?an?effective?triggering?and?the?variation?span?of?Pa?during?the?triggering?period?were?analyzed?in?respiratory?mechanics.? Results? ①?In?flow-trigger?mode:?Pa?was?pulled?down? from? positive? end-expiratory? pressure? (PEEP)? or? intrinsic? positive? end-expiratory? pressure? (PEEPi)? to?"PEEP－VTrigR"?(R?meant?airway?resistance)?without?nebulization,?and?the?span?of?Pa?was?"VTrigR"?or?"PEEPi－PEEP+VTrigR".?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－(VTrig+N)?R"?(N?meant?nebulization?airflow)?with?nebulization,?and?the?span?of?Pa?was?"(VTrig+N)?R"?or?"PEEPi－PEEP+(VTrig+N)?R".?②?In?pressure-trigger?mode:?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－PTrig－1R"?without?nebulization,?and?the?span?of?Pa?was?"PTrig+1R"?or?"PEEPi－PEEP+PTrig+1R".?Pa?was?pulled?down?from?PEEP?or?PEEPi?to?"PEEP－PTrig－(N+1)?R"?with?nebulization,?and?the?span?of?Pa?was?"PTrig+(N+1)?R"?or?"PEEPi－PEEP+PTrig+(N+1)?R".? ?Conclusions? Nebulization?airflow?increases?the?difficulty?of?inspiratory?triggering?in?mechanical?ventilation.?PEEPi?makes?it?more?difficult.
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Objective To design a mathematical calculation model for better understanding and grasping the logical problem of replacement fluid and citric acid anticoagulant infusion in continuous veno-venous hemofiltration (CVVH). Methods ① Parameter definition: A, B, and T were respectively called the main part of pre-replacement fluid, 5% sodium bicarbonate solution, and 4% sodium citrate infused before filter. And a and b were respectively called the main part of post-replacement fluid, and 5% sodium bicarbonate solution infused after filter. ② Logic conversion:The liquid in back terminal (Z) was artificially divided into two parts. One (X) was the original residual plasma after filtration. The second (Y) was the part excluding the plasma, including the left part of pre-replacement fluid with sodium citrate, and the post-replacement fluid. ③The mathematical formulas of liquid volume and electrolyte concentration at X, Y and Z in unit time were listed according to the principle of CVVH and the screening coefficient of filter for different substances. ④The calculation formulas were entered into Excel form, and a mathematical calculation model was made, and a simulation calculation with examples was carried out. Results An Excel model was established by inserting the calculation formulas of volume, electrolyte, and total calcium at X, Y and Z. And it was found that the concentration of Na+, K+, Cl-, HCO3- at Y point remained unchanged only when A, B and (or) a, b was kept in same side and proportion even with the change of blood flow and other parameters without sodium citrate as anticoagulant. Once any of the parameters (such as blood flow, replacement fluid volume, etc.) were adjusted in other infusion methods (such as different ratios, different directions of the same year, etc.), the calculation results at Y would vary, and the electrolyte concentration at Z would change accordingly. A change of dilution model or parameter would result in the change of the electrolyte concentration at Y and Z with sodium citrate as anticoagulant. The concentration of total calcium scarcely changed no matter in what model and parameters. Conclusions All kinds of infusion ways could be included in the Excel model. The infusion results of all kinds of infusion matching could be intuitively evaluated. It is helpful for the medical staff to make a logical analysis and risk prediction in CVVH.
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Objective To prove with mathematical formula that the patient's blood electrolyte concentration shows a tendency to approach that of replacement-fluid after continuous renal replacement therapy (CRRT).Methods Electrolyte concentration of plasma,replacement-fluid and returning fluid were compared,and they were labeled as Cblood,Cnom,and Creturn respectively.The Creturn was calculated,and the relationship among them was demonstrated with comparison by mathematical formula.At last,according to their relationship,plasma change towards to the replacement fluid was analyzed.Results It was showed that Cblood＜Creturn＜Cnom or Cblood＞Creturn＞ Cnom,and according the relationship,it was derive that the trend of change in Cblood after circulation for m unit time was Cblood1 ＞Cblood2 ＞Cblood3 ＞ … ＞Cbloodm ＞Cblood or Cblood1 ＜ Cblood2 ＜Cblood3 ＜ … ＜Cbloodm ＜Cnom.The plasma electrolyte concentration would close to that of replacement fluid infinitely with the continue of CRRT.Conclusions With mathematical model,it is proved that the replacement fluid electrolyte concentration is the final target of the plasma.We must make up the replacement fluid correctly.And this results provide the basis for CRRT treatment of electrolyte disorder.
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Objective To study the clinical effect of large dose ulinastatin on patients combined SIRS and MODS.Methods 120 patients in ICU were randomly divided into two groups,each 60 cases,the control group using conventional standard treatment,the observation group in the control group on the basis of the 12h after the operation started on intravenous injection of large dose ulinastatin,observe two groups of patients after treatment with cytokine content and duration of hospital stay,mortality rate and APACHE Ⅱ score after treatment.Results The two groups before treatment TNFα,IL-6,IL-8 and APACHE Ⅱ score difference is not big,after treatment,48 h,72 h,two groups of patients with TNFα,IL-6,IL-8 were decreased,and the observation group decreased greater than the control group (t =8.46,8.72,8.68,10.38,7.83,9.26,all P ＜ 0.05),after treatment the observation group APACHE Ⅱ score,mortality,average time of hospitalization were significantly lower than those of the control group,there was statistical significance(t =9.58,x2 =10.28,t =9.17,all P ＜ 0.05).Conclusion High dose ulinastatin on inflammatory cytokines have obvious inhibition effect,can reduce the surgical risk in patients with APACHE Ⅱ score and mortality,shorten the treatment time of patients.
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Objective To assess the impact of the virus on the complementary determining region 3 (CDR3) length diversity of T cell receptor(TCR) Vβ repertoires of CD4+ T lymphocytes and to explore its association with viral load in individuals with HIV-1 infection. Methods The TCR repertoire was examined using spectratyping of CDR3 length diversity within CD4+ T cells in HIV infected and healthy adults. Separation of CD4+ T cells from peripheral blood mononuclear cells ( PBMCs) was carried out by using immunomagnetic beads coated with anti-CD4 antibody. Total RNAs from the purified CD4 + T lymphocytes were isolated and used to perform nested-PCR amplifications in CDR3 of 22 TCR gene families. CDR3 diversity and its association with viral load in individuals with HIV-1 infection were analyzed. Results An average diversity for all CDR3 profiles in CD4+ T cells from 25 HIV-infected individuals was significantly different as compared to 10 age-matched healthy donors (P＜0.05) with the HIV-infected individuals losing diversity in the CDR3 profiles. There was positive correlation between changes in TCR CDR3 diversity and viral load (r = 0. 494, P ＜ 0. 05). The changes in CDR3 length diversity of Vβ families in HIV-infected individuals, particular in Vβ8, Vβ22, Vβ23 were statistically different from the healthy controls. Conclusion HIV-1 infection might induce the loss of TCR Vp repertoire diversity and disrupt the CDR3 Gaussian distributions within CD4 + T cells. There should be positive correlation between changes in TCR CDR3 diversity and the viral load in HIV-1 infected patients.
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Objective To explore the polymorphism of HLA class I alleles of HIV-infected former plasma donors and to investigate its impact on HIV-1 viral load in central China.Methods 106 subjects chronically infected with HIV-1 were recruited and HLA class I alleles were genotyped with PCR-SSP assay.HLA class I genotypes and haplotypes were determined and their association with plasma viral loads were analyzed.Gag-specific CTL responses were detected by an IFN-λ EUSPOT assay by using overlapping peptides,and their association with plasma viral loads were also analyzed.Results Subjects homozygous at HLA class I locus had higher plasma viral loads(P=0.0098);HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads(P=0.0004,0.0103,0.0058,0.0371 and 0.0006);an inverse correlation between p2p7p1p6-specific CTL responses and viral loads in subjects with HLA-A*30/B*13/Cw*06 haplotype as well as an inverse correlation between p17-specific CTL responses and viral loads in subjects with HLA-Cw*14 allele were observed.Conclusion HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads and Gag-specific CTL responses restricted by these HLA alleles may contribute to the protection.
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Objective To explore the impact of broad antigen HLA-Bw4 on disease progression in HIV-1 infected subjects. Methods Three hundred and forty subjects chronically infected with HIV-1 and 69 HIV-1 negative subjects were recruited and HLA-B alleles were typed with sequence-based high resolution typing assay. HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD4+ T cell counts and viral loads were analyzed. Results Sixty-five HLA-B alleles were detected in HIV-1 positive subjects. Subjects with Bw4 (Bw4 homozygotes and Bw4Bw6 heterozygotes ) had higher CD4+ T cell counts ( P = 0. 004 ) and lower plasma viral load ( P = 0.003 ) than subjects without Bw4 ( Bw6 homozygotes). When compared with HIV-1 postive subjects with CD4+ T cell counts above 500 celis/μl, those with CD4+ T cell counts below 500 cells/μl were observed with decreased percentage of Bw4Bw6 heterozygote ( P =0.0002) and increased percentage of Bw6 homozygotes ( P < 0. 0001 ). There is no significant difference in CD4+ T cell counts between Bw4 homozygotes and Bw4Bw6 heterozygote, but lower viral loads were observed in Bw4Bw6 heterozygotes( P = 0. 037 ). Conclusion HLA-Bw4 can confer pretective effects on H1V-1 infected subjects by maintaining higher CD4+ T cell counts and lower viral load, the mechanism behind this effect need further exploration.
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Objective To provide an anatomic data of the lumbar isthmus, and to increase the fusion rate of the graft bone for spondylolisthesis treated with posterolateral spinal fusion and Steffee plate fixation. Methods The area and volume of the bone graft bed for posterolateral spinal fusion in the lumbar isthmus were studied on 30 dried specimens of adult lumbar spines from L 1 to L 5(a total of 150 lumbar vertebrae) with a vernier calipers, a goniometer and a graduate. Results The length of the superior edge of the isthmus gradually increased from L 1[(7.34? 1.15) mm] to L 5[(10.31? 1.89) mm], and that of its inferior edge progressively decreased from L 1[(6.63? 1.24) mm] to L 5 [( 4.73? 1.21) mm]. The superior edge of the isthmus was the thinnest at L 1[(1.85? 0.64)mm], and the thickness increased inferiorly from L 1[(6.57? 1.24) mm] to L 5[(7.57? 1.16) mm]. The medial and caudal inclination of the isthmus with respect to the pedicle gradually increased from L 1 (113.9?? 13.3? ) to L 5(119.1?? 13.5? ) medial inclination and from (101.2?? 9.5? ) at L 1 to (117.6?? 13.4? ) at L 5 caudal inclination, respectively. The length of the superior edge of the isthmus and the transverse diameter of the pedicle were positively correlated to the vertebral levels. The length of the inferior edge of the isthmus were negatively correlated to the vertebral levels. The dimensions of the graft bed area were postively correlated to dimensions of the graft bone volume. Conclusion The application of firm spinal internal fixation system with adequate graft bed area and adequate graft volume, the maintenance of close contact between the graft bed and the graft bone, and adequate period of postoperative stable contact of the graft in bed were the keys to enable reduction of the spondylolisthesis and to increase the fusion rate of posterolateral bone graft.
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The arterial supply of the human metatarsals in 90 lower limbs,ranging in age from newborn to87 years,was studied by perfusion method. The blood supply of the metatarsals is derived from the nutrient artery,the periosteal arteries,the epiphyseo-metaphyseal arteries and the pseudoepiphyseal arteries. The proximal and distal branches of the nutrient artery supply the inner two-thirds or three-fourths of the cortex. The periosteal arteries give off a few twigs penetrating into the cortex and supplying the outer one-third or one-fourth of the cortex. The radiate form epiphyseo-metaphyseal arteries derived from superficial plexuses in continuity with the periosteal networks on the shafts, supply the base of the first metatarsal, and the heads of the other four. The pseudoepiphyseal arteries, penetrating the non-articular areas of the bones, supply the bases of the second, third, fourth and fifth metatarsals and the head of the first. The sources of the nutrient arteries to the metatarsal vary with the different bone . The nutrient arteries to first metatarsal bone usually originate from the ramus plantaris profundus and the first plantar metatarsal artery, while the nutrient arteries to the other metatarsal bones mainly derive from the plantar metatarsal arteries, the plantar arch and its perforating branches. The clinical significance of the arterial distribution to the metatarsals is discussed.
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Microvascular architecture of the oviduct was observed by SEM in 20 adult female Wistar rats which were not pregnant. The oviduct was supplied by 2-4 tubal branches (0.2-0.3mm in diameter) which were derived from the ovarian artery. The tubal branches entered the wall of oviduct and divided into numerous meandering arterioles (0.02-0.07mm in diameter). Intrinsic microvasculature of the oviduct might be divided into three layers: (1) the subserosal vascular plexus which appeared tortuous and formed reticulate anastomosis, (2) a parallel branching vasculature in the muscle coat, and (3) the subepithelial capillary network. The latter arised from the arterioles which ramified terminally near the apex of mucosal folds and broke up into very dense capillaries. Subepithelial capillary density in the fimbriae was markedly higher than that in the isthmus and uterine part. Intrinsic venules of oviduct were less tortuous and less closely accompanied with the arterioles. In mesoviduct, veins were in closely accompanied with the corresponding arteries. In the wall of oviduct, some capillaries and venules drained to the small veins directly and vertically.
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Objective To detect the expression and correlated significance of inducible nitric oxide synthase and p53,Bax in benign and malignant diseases of gallbladder. Methods In 16 cases of chronic cholecystitis,11cases of chronic cholecystitis with adenomyoma and 24 cases of gallbladder adenocarcinoma,the expression of iNOS,p53 and Bax in gallbladder's wall was detected by SP immunohistochemistry. Results 1