Objective To study the effect of hypothyroidism on oxidative stress status in developing rat brain and to further explore the mechanism of impaired brain development caused by hypothyroidism. Methods Perinatal hypothyroidism was induced by administering propylthiouracil(PTU) solution to the dams by gavage.The oxidative stress indexes were measured in brain homogenate of normal and hypothyroid pups which were sacrificed on the 21st d after birth. Results As compared to the control,the following indexes were found to be increased in the hypothyroid group: protein carbonyl contents,thiobarbital acid reactive substances,reduced glutathione,total antioxidative capacity,activities of superoxide dismutase and glutathione peroxidase(P0.05). Conclusion Hypothyroidism during rat brain development may cause oxidative stress,which may be related to the brain damage caused by hypothyroidism.

AlM:To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes.
METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3. 1 +, and accredited whether pcDNA3. 1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3. 1+/TSHR289.
RESULTS: Recombination plasmid pcDNA3. 1+/TSHR289 digested with enzyme Hindlll and the fragment through 0. 8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3. 1+/TSHR289 were found synonymous mutation through forward ( AAC to AAT ) and reverse sequencing ( GCG to GCT) . The volume ratio of cationic liposomes and recombinant plasmid was 3:1.
CONCLUSlON: lt is successful to construct the recombination plasmid pcDNA3. 1+/TSHR289 by accredit it through enzymatic digestion and sequencing.

Objective To explore the preventive and therapeutic role of silencing type Ⅰ rat platelet-binding protein motifs depolymerization protein-like metalloproteinase 2(ADAMTS2)by siRNA on experimental liver fibrosis in vitro.By studying the mechanism of siRNA silencing of ADAMTS2,we also aim to evaluate the feasibility of ADAMTS2 as a target for anti-liver fibrosis therapy.Methods Three pairs of siRNAs targeting ADAMTS2 mRNA 2237,2597 and 690 targets were designed and synthesized by utilizing RNA design software.The most effective siRNA was chosen to transfect HSC-T6 cell line to test the tendency of hepatic stellate cell(HSC)activation and ex pression of ADAMTS2,COL1α1,COL(I),α-SMA,TGF-β1,MMP-2 and TIMP-3.These were quantified using real time-PCR,Western blotting,and MTT assays.Results Of the same dosage and time of injection,siRNA 2237 inhibited ADAMTS2 gene expression significantly more than other siRNAs.siRNA-ADAMTS2 2237 markedly inhibited ADAMTS2 gene and protein expression of HSCT6 with more than 80％ efficiency.Conversely,siRNA-ADAMTS2 2237 markedly reduced the gene and protein expressions of COL(I),α-SMA and TGF-β1 on HSC-T6 and inhibited the proliferation of HSC.Conclusions siRNA-ADAMTS2 2237 could effectively knockdown the gene and protein expression of ADAMTS2 in HSC-T6 cell lines.Silencing ADAMTS2 by siRNA significantly inhibited the activation,proliferation of HSC and the gene and protein expressions of COL(I),α SMA,and TGF-β1,and it may have a potential anti-fibrotic effect.ADAMTS2 might be an efficient target for anti-fibrotic therapy.

Objective To explore the enhancement patterns of pulmonary carcinomas by contrast-enhanced ultrasound(CEUS).Methods Thirty-eight patients with pulmonary carcinomas proven by pathology[28 with peripheral pulmonary carcinomas and 10 central pulmonary carcinoma with obstructive atelectasis(OA)]were examined by baseline ultrasound and contrast-enhanced ultrasound,then the arrival time(AT),time to peak(TTP)were analyzed with time-intensity curve analysis software and the dynamic enhancement pattern of each lesion was assessed.Results Twenty-four peripheral pulmonary carcinomas demonstrated delayed AT about 6-16 s after application of contrast medium,three lesions demonstrated early AT about 4-5 s and one lesion demonstrated absence of contrast enhancement.The lesions exhibited hyper-,hypo- and non-enhancement were 14,13 and 1,respectively.Seventeen lesions were heterogenous enhanced with non-enhanced necrosis areas and enhanced septa,while ten lesions homogeneous enhanced and one lesion no enhanced.Ten central pulmonary carcinoma with OA demonstrated a characteristic pattern:OA appeared a short AT(mean AT 4.8 s)until enhancement and strong contrast enhancement,while the central tumors appeared a delayed AT(mean AT 10.5 s)and faint enhancement.Conclusions CEUS can be useful in differentiation between solid and cystic pulmonary lesions,and detection of the latent lesions underlying the atelectasis.

Enhancing medical students' humane quality education is an urgent requirement for modem medical mode transformation for medical education.The pharmacology teachers of Chongqing Medical University follow the modem education concepts and fully search the human spirit materials hidden in pharmacology,then actively explore how to integrate the humanity spirit education into the pharmacology teaching to achieve the changes of from exam-oriented education to quality education.

The study was purposed to analyze the frequency and distribution of irregular antibodies in Shaoguan area. Screening 15 033 random donor antibodies in Shaoguan area by screening cells, polybrene and antiglobulin tests. The results indicated that the irregular antibodies were found in 42 samples. The frequency of irregular antibodies in female was higher than that in male (P < 0.001), and Rh blood group antibodies such as anti-D, anti-E, and anti-Ec C were common (47.6%). 2 samples of Le antibodies were failed to be found by polybrene test. 2 samples of irregular antibodies with titer 2 were undiscovered by screening test of 10 pooled samples. In conclusion, because of irregular antibodies resulting in hemolytic transfusion reaction, the investigation of frequency and distribution of irregular antibodies is very important for safe transfusion. Antibody screening must be done for female donors, and especially for massive plasma transfusion of patients with severe and dangerous illness and infants so as to ensure safety.
Adolescent ; Adult ; Blood Donors ; China ; Erythrocytes ; immunology ; Female ; Humans ; Isoantibodies ; blood ; Male ; Mass Screening ; Middle Aged ; Rh-Hr Blood-Group System ; blood ; immunology ; Rho(D) Immune Globulin

OBJECTIVETo search for the optimal strategies for sperm collection from the patient with temporary penile erectile dysfunction (ED) on the day of oocyte pick-up ( OPU) in in vitro fertilization embryo transfer (IVF-ET).

METHODSWe retrospectively analyzed 93 cases of temporary ED on the OPU day of IVF-ET from January 2011 to May 2014, with fresh semen for 45 cases (group A), cryopreserved sperm before oocyte retrieval for 30 cases (group B), and frozen oocytes for 18 cases (group C). Group A was again subdivided into A1 (n = 18) and A2 n = 27) , the former intervened with oral sildenafil while the latter left untreated. We compared the rates of fertilization, high-quality embryo, and pregnancy among different groups.

RESULTSNo statistically significant differences were found among groups A, B and C in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and rates of cleavage, or in the percentages of normal fertilization (80.78% vs 80.43% vs 84.77%), high-quality embryo (53.27% vs 52.97% vs 47.69%) and pregnancy (60.00% vs 56.77% vs 44.44%) (all P > 0.05). The rate of 3PN was markedly lower in group C (0.63%) than in A (9. 61%) and B (4.34%) (P < 0.05). There were no significant differences between groups A1 and A2 in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and the rates of fertilization, cleavage, high-quality embryo, and pregnancy (all P > 0.05).

CONCLUSIONOn the OPU day of IVF-ET, oral sildenafil can help temporary ED men to achieve penile erection and ejaculation without affecting the outcomes of assisted reproduction. Cryopreserved sperm can be used in case of predicted temporary ED and frozen oocytes can also be employed if sperm retrieval fails. However, to avoid puncture injury to the epididymis or testis, fresh semen should be the first choice.

Cryopreservation ; Embryo Transfer ; Erectile Dysfunction ; drug therapy ; physiopathology ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Male ; Oocyte Retrieval ; Oocytes ; Penis ; Pregnancy ; Retrospective Studies ; Sildenafil Citrate ; therapeutic use ; Sperm Retrieval ; Spermatozoa

Objective To construct a lentiviral vector-based short hairpin RNA (shRNA) targeting the gene encoding tryptophan-aspartate containing coat protein ( TACO) and to evaluate its inhibitory effect on the expression of TACO , and to further elucidate its effects on the phagocytosing and intracellular killing of My-cobacterium tuberculosis (M.tb) by macrophages and the possible mechanisms.Methods Three shRNA frag-ments targeting TACO gene and a scrambling control shRNA fragments were designed and cloned into the lentivi -ral vector pSicoR .The recombinant lentiviral vectors were identified by sequencing analysis and then packed in 293T cells.Real-time RT-PCR and Western blot assay were performed to evaluate the gene-silencing efficiency of the recombinant lentiviral vectors among RAW 264.7 cells transfected with the concentrated lentivirus .The most effective lentivirus was screened out to transfect the RAW 264.7 cells for 48 hours, followed by infection those cells with M.tb strains.The entry and intracellular survival of M .tb strains in RAW264.7 cells were de-termined by bacterial culture at indicated time points .The colocalization of M .tb and lysosomes was detected by immunofluorescence staining .The cyto-ID autophagy kit was used to detect the cellular autophagy and the auto-phagy-associated protein LC 3 was determined by Western blot assay .Results The recombinant lentiviral vec-tors were successfully constructed and confirmed by sequencing analysis .Decreased expression of TACO in RAW264 .7 cells was detected after transfecting the cells with the lentiviral vector-based shRNA vectors targeting TACO gene for 48 hours.The most effective lentivirus , LV-pSRT1, decreased the expression of TACO by 85.24%and 69.00%at the mRNA and protein levels, respectively.The bacterial loads in LV-pSRT1 trans-fected RAW264.7 cells were significantly decreased at the time point of 0 h after M.tb infection as compared with those in the control lentivirus treated cells (5.50×104 vs 8.1 ×104, P<0.05).Compared with the RAW264 .7 cells transfected with control lentivirus , the survival rate of intracellular M .tb strains in LV-pSRT1 transfected cells was significantly decreased at the time point of 48 h (134.54% vs 213.58%, P<0.05) and 72 h (148.18%vs 262.96%, P<0.05) considering the bacterial load at the time point of 0 h as the standard. The immunofluorescence staining demonstrated that the colocalization of M .tb strains with lysosomes was signifi-cantly enhanced in LV-pSRT1 transfected cells as compared with that in control lentivirus treated cells (75.67%vs 10.66%, P<0.05).Moreover, significantly enhanced autophagy and relative expression of LC 3Ⅱ protein were observed in RAW264.7 cells with TACO gene knockdown (16.20%vs 8.50%, P<0.05;0.51 vs 0.34, P<0.05).Conclusion The lentiviral vector-based shRNA targeting TACO gene could effectively knockdown the expression of TACO protein , decrease the entry and increase the intracellular killing of M .tb strains in mac-rophages.The enhanced intracellular killing of M .tb strains by macrophages was associated with the increased fusion of M.tb-containing phagosome and lysosome .

· AIM: To report a rare case of mesenchymalchondrosarcoma in the orbit and to explore its clinicmanifestations, pathologic characters, management andprognosis. · METHODS: We report a case of mesenchymalchondrosarcoma of the orbit. The clinical materials,including ophthalmological examination, computed tomo-graphy scan of the orbit, histopathology and immunohis-tochemistry of the biopsy specimen was reported, and itspertinent literatures were reviewed.· RESULTS: A 36-year-old female was seen with proptosisand decreased vision. Histopathology demonstrated anadmixture of undifferentiated mesenchymal cells andislands of mature hyaline cartilage. Immunohistochemicalstudies revealed positivity for vimentin and S-100, whichwas consistent with the diagnosis of mesenchymalchondrosarcoma.· CONCLUSION: Mesenchymal chondrosarcoma in theorbit is extremely rare malignant tumor. Multi-modalitytreatments (surgery, chemotherapy and radiotherapy)may lead to long-term survival.

· AIM: To investigate whether heat shock protein 27 (HSP27) is induced in retinal ganglion cells (RGCs) in experimental rat glaucoma and whether the induction of HSP27 by intraocular pressure (IOP) elevation can increase serum autoantibody to HSP27 in the model.IOP elevation、Sham and normal groups by SPSS12.0. IOP was raised by electrocoagulating at least 3 episcleral veins and limbal veins on the right eye of each rat in IOP elevation group and its contralateral eye was used as controls. Immunohistochemical staining for HSP27 was performed in RGCs and retinal nerve fiber layer (RNFL) and serum immunoreactivity against HSP27 was detected by means of enzyme-linked immunosorbent assay (ELISA) in three groups.RNFL of the eyes with IOP elevation, while it was expressed weakly in untreated control eyes. Compared with sham and normal groups, serum autoantibody to HSP27 was slightly high at 1wk (P ＞0.05) and significantly increased at 2, 3, 4 and 8wk (P＜0.05) in IOP elevation group.enhanced expression of the endogenous HSP27 might play an important role in glaucomatous optic neuropathy.