Objective To observe the clinical characteristics and efficacy of laser photocoagulation of aggressive posterior retinopathy of prematurity (AP-ROP).Methods Twenty-eight eyes of 14 infants with AP-ROP from May 2008 to December 2010 were enrolled in this study.The infants were examined using RetCam photography and indirect ophthalmoscopy.Among the 28 eyes,24 eyes were classified as zone one and 4 eyes zone two.All eyes were treated within 24 hours using binocular indirect ophthalmoscope and laser photocoagulation.The initial energy was 200 mW,using energy was 200 - 500 mW,exposure time was 200 ms.Every two photocoagulation spot was linked together,but no overlap.Follow-up ranged from 3 to 24 months,with a mean of 11.5 months. The retinal bloods,the iris surface vessels,the fiber hyperplasia on retina,retinal detachment or ruffle form were observed.Results Twenty-five of 28 eyes (89.3 ％) recovered or were classified as control; 1 of 28 eyes (3.6 ％) was suffered retinal detachment one month after treatment.The detachment was resolved through vitrectomy Surgery.Two of 28 eyes (7.1 ％)did poorly.The parents gave up treatment resulting in loss of vision.No treatment-related complications were observed during the follow-up period,such as damage to cornea,iris and lens. Conclusion Photocoagulation is a safe and effective way to treat most AP-ROP.

Objective To describe the clinical feature ,therapeutic approach and prognosis of histoplasmosis for improving clinicians’ awareness of this disease .Methods The clinical data of 7 cases of histoplasmosis treated in Shanghai Huashan Hospital from 2001 to 2014 were reviewed retrospectively .Relevant reports about histoplasmosis from 2001 to 2014 in Chinese mainland were comprehensively reviewed .Results The major clinical manifestations of progressive disseminated histoplasmosis included fever ,hepatosplenomegaly ,lymphadenopathy ,and pancytopenia .Skin lesions and pancytopenia were more common in the patients complicated with HIV/AIDS .The patients with local infection were lack of systemic symptoms or signs . Histological examination found Histoplasmacapsulatum in macrophages in bone marrow or biopsy tissues .Amphotericin B was used most frequently to treat histoplasmosis .Itraconazole was appropriate in mild patients .Conclusions Histoplasmosis is caused by H .capsulatum .The golden standard of diagnosis is any culture positive for H .capsulatum .Antifungal treatments such as amphotericin B and itraconazole are very important .

Objective:To express SARS-CoV nucleocapsid protein in Bac-to-Bac Baculovirus Expression System and analyze the antigenicity of the recombinant protein.Methods: The SARS-CoV nucleocapsid gene was amplified by PCR.The PCR product digested with BamHⅠand SalⅠrestriction endonucleases was cloned into vector pFastBac HTC of Bac-to-Bac Baculovirus expression system.Recombinant plasmid was transformed DH 10Bac cells to obtain the recombinant Bacmid DNA.Recombinant Bacmid DNA was transferred into Sf9 cells which were inducted to express the recombinant protein in High Five cells.After purified by Ni affinity chroma-tography ,the antigenicity of the recombinant protein was analyzed by Western blot and ELISA.Results:Recombinant plasmid was con-structed successfully.The recombinant protein with the relative molecular mass of 48 kD was efficiently expressed in High Five cells and purified successfully by Ni affinity chromatography.Western blot and ELISA analysis showed that the recombinant protein could be spe -cifically recognized by the monoclonal antibody to SARS-CoV N protein and immune serum from rabbits ,respectively.The recombinant protein can specifically reacted with serum from SARS patients ,not with serum from healthy persons and patients infected with hCoV-229 E and hCoV-OC43.Conclusion: SARS-CoV nucleocapsid protein has been expressed successfully in the Bac-to-Bac Baculovirus Expression System ,and obtained good antigenicity.It is preliminary deemed that it can't reacted with serum from patients infected with hCoV-229E and hCoV-OC43.

The present study is to explore the change process and distribution of phosphorylated DARPP-32 (p-DARPP-32) in rat brain including cortex, hippocampus and striatum and to further deduce whether p-DARPP-32 was possibly involved in epilepsy induced by repetitive low doses of pentylenetetrazol (PTZ). PTZ-induced epilepsy model in rat was established with 30 male SD rats randomly divided into 6 groups, control group and five trial groups [PTZ 1 h, PTZ 6 h, PTZ 24 h, PTZ 48 h and PTZ 72 h respectively, after onset of status epilepticus (SE)]. Immunohistochemistry and immunofluorescence double-labeling were used to detect the temporal time change and distribution of p-DARPP-32 expression and to analyze the coexpression of DARPP-32 and p-DARPP-32 in rat brain after the onset of PTZ-induced generalized SE. The results showed that there was a temporal time change of p-DARPP-32 expression in rat brain after the onset of SE. The number of p-DARPP-32-positive cells increased significantly and reached the peaks at the ends of 1 hour and 6 hours after the onset of SE, but decreased at the end of 24 hours. The moderate to strong p-DARPP-32-immunopositive neurons were observed in cortex, hippocampus and striatum, and located in cell cytoplasm and cell nucleus. Further immunofluorescence double-labeling revealed that denser colocalization of p-DARPP-32 and DARPP-32 in the neurons existed in the area mentioned above. Therefore, PTZ-induced SE may cause phosphorylation of DARPP-32 in rat brain. The temporal time change and distribution of p-DARPP-32 suggest that phosphorylation of DARPP-32 may be involved in PTZ-induced epilepsy in rat brain including cortex, hippocampus and striatum, and p-DARPP-32 may play a central role in the onset of SE.
Animals ; Cerebral Cortex ; metabolism ; Corpus Striatum ; metabolism ; Dopamine and cAMP-Regulated Phosphoprotein 32 ; metabolism ; Hippocampus ; metabolism ; Male ; Neurons ; metabolism ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; metabolism

Objective To study the effects of oral rehydration with the solution of pyruvate-glucoseelectrolyte (PGES) by comparison with the bicarbonate-glucose-electrolyte solution (BGES) on resuscitation in rats with lethal hemorrhagic shock.Methods Sixty adult male SD rats with intra-gastric tube,and cannulation of femoral artery and vein were subjected to 45％ total blood volume loss from the femoral artery,and then randomly divided into three groups (n =20 in each group):no fluid resuscitation group (NR),oral fluid resuscitation with the PGES group (PGES) and oral fluid resuscitation with the BGES group (BGES).In NR group,the animals received no fluid replacement or any other treatment.Rats in PGES and BGES groups were infused intra-gastrically with pre-warmed PGES or BGES in volume of 2 times shed blood given at 30 min after hemorrhage and completed within 6 hours.Blood samples in each group were collected from the abdominal aorta before or at 0,1,2,4 h post hemorrhage to detect serum alanine aminotransferase (ALT),creatinine (Cr),creatine phosphate kinase isoenzyme (CK-MB) and intestinal fatty acid binding protein (iFABP).Another 84 rats randomly divided into four groups:NR group (n =24),PGES group (n =24),BGES group (n =24),and no hemorrhage group (NH group,n =12).Rats in the three hemorrhage groups were treated the same as described above,and the rats in NH group underwent the same surgical procedure without hemorrhage were served as the sham group.All these rats were observed for their 24-hour survival rates.Results The 24-hour survival rates of PGES and BGES groups were both significantly higher than the rate of NR group (11/24 vs.1/24,x2 =18.087,P ＜0.01 ; 5/24 vs.1/24,x2 =6.445,P ＜ 0.05) ; the survival rate of PGES group was also significantly higher than that of BGES group (11/24 vs.5/24,x2 =4.02,P ＜ 0.05).All levels of ALT,CK-MB,Cr and iFABP in both the NR group and two oral resuscitation groups at 1 h,2 h and 4 h post hemorrhage were significantly higher than those before the blood loss,respectively (P ＜ 0.01).These biomarkers at 2 h,4 h post hemorrhage were significantly lower in the PGES and BGES groups than those in NR group (P ＜ 0.01) ; the serum levels of ALT,CK-MB,Cr and iFABP were significantly lower in the PGES group than those in the BGES group at 2 h and 4 h post hemorrhage,respectively (P ＜ 0.05).Conclusions Present results demonstrated that the pyruvate-enriched oral re-hydration solution (ORS =PGES) was more effective in preserving the organ function and prolonging the animal survival after resuscitation of lethal hemorrhagic shock in comparison with the bicarbonate-containing ORS (BGES).The oral re-hydration solution (PGES) recommended by the World Hygiene Organization (WHO ORS) may require further improvement in oral resuscitation of shock and the PGES may be recommended as a choice of oral re-hydration salts in the treatment of lethal hemorrhagic shock when intravenous administration is not available.

Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; immunology ; Dengue ; virology ; Dengue Virus ; chemistry ; genetics ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Viral Envelope Proteins ; chemistry ; genetics ; immunology

OBJECTIVE To study the etiological diagnosis of invasive aspergillosis by the monoclonal antibodies against Aspergillus fumigatus. METHODS An animal model of rabbit invasive aspergillosis was established.The antigen of A.fumigatus in serum was detected by ELISA.The antigen of A.fumigatus in tissue was detected by immunochemistry. RESULTS ELISA assay showed positive 24,48 and 72 hours after infection.Immunochemistry was positive 72 hours after infection. CONCLUSIONS The monoclonal antibodies against A.fumigatus has great potency usage.

Objective To study the anatomic data of the first metatarsal dorsal artery and to provide anatomical basis for clinical tissue transplantation based on the first metatarsal dorsal artery.Methods The 16 adult cadaver specimens with 32 feet were dissected and meas-ured by vernier caliper.Then the anatomic data of the first metatarsal dorsal artery were analyzed.Results Through the examinations of 32 feet sample,the first metatarsal dorsal artery were classified into 5 types.Type Ⅰ:the first metatarsal dorsal artery runs at the surface of the first dorsal interosseous muscle (13 sides,40.6%).Type Ⅱ:the first metatarsal dorsal artery runs in the interior of the first dorsal interosse-ous muscle (11sides,34.4%).Type Ⅲ:the first metatarsal dorsal artery runs underneath the first dorsal interosseous muscle (6 sides, 18.8%).Type Ⅳ:the first metatarsal dorsal artery is slender (1 side,3.1%).TypeⅤ:the first metatarsal dorsal artery is absent (1 side, 3.1%).Distance relationship was measured between the first metatarsal bone and the first metatarsal dorsal artery:the vertical distance be-tween the origin of the posterior branch of the first metatarsal dorsal artery and base of the first metatarsal bone was (2.4 ±0.3)mm,the ver-tical distance between the origin of the posterior branch of the first metatarsal dorsal artery and head of the first metatarsal bone was (10.1 ±1.0)mm;the vertical distance between the origin of the anterior branch of the first metatarsal dorsal artery and the first metatarso-phalangeal joint was (7.6 ±2.7)mm.Conclusion The first metatarsal dorsal artery has clinical reference significance for the hands and feet’s trauma and skin flap transplantation such as thumb reconstruction.

OBJECTIVE: To explore the molecular basis for a pedigree affected with spondyloepiphyseal dysplasia congenita (SEDC). METHODS: The proband was subjected to whole exome sequencing. Suspected variant was verified by Sanger sequencing. RESULTS: All patients from the pedigree were found to carry a novel missense variant c.1394G>C (p.Gly465Ala) of the COL2A1 gene. The variant was not reported previously. Provean, Polyphen-2 and Mutation Taster software predicted that the variant is highly likely to be pathogenic. CONCLUSION The c.1394G>C (p.Gly465Ala) variant of the COL2A1 gene probably underlies the SEDC in this pedigree.
Asian Continental Ancestry Group ; Collagen Type II ; genetics ; Humans ; Osteochondrodysplasias ; congenital ; genetics ; Pedigree

Objective:To explore the effects of G protein-coupled receptor 55 (GPR55) antagonist CID16020046 on renal fibrosis in mice, and provide a new method and idea for the treatment of renal fibrosis.Methods:(1) GPR55 overexpression and GPR55 antagonist CID16020046 were used in renal fibroblasts (NRK-49F) of rats, respectively. Meanwhile,transforming growth factor-β1 (TGF-β1) was applied in the NRK-49F cells to observe the expression of fibrosis-related factors and inflammatory factors. (2) A mouse model of renal fibrosis with unilateral ureteral obstruction (UUO) was established in vivo. Eight-week-old male C57BL/6J mice (20-25 g) were randomly divided into three groups according to the random number table method: sham group ( n=6), model group (UUO group, n=7), model + CID16020046 drug (UUO+CID group, n=8). The drug CID16020046 (10 mg/kg) was intraperitoneally injected 1 day before modeling, on the day of modeling and every day after surgery in UUO+CID group, and the corresponding dose of 0.9% normal saline was injected intraperitoneally in sham and UUO groups.The mice were sacrificed for sampling 7 days after UUO surgery, and their renal function indicators, liver transaminase, and cardiac markers were examined. Western blotting and quantitative real-time PCR were used to detect the expression of renal fibrosis-related factors and inflammatory factors. Immunohistochemistry staining, Sirius red staining and Masson trichrome staining were used to detect the pathological changes of renal tissues. Results:(1) After NRK-49F cells were stimulated by TGF-β1, the mRNA and protein expression levels of GPR55 were significantly increased (both P<0.05). There was no statistically significant difference in the mRNA expression of fibrosis-related factors fibronectin and collagen Ⅰ, and inflammatory factors interleukin-1β and tumor necrosis factor-α between TGF-β1 group and TGF-β1 + GPR55 overexpression group (all P>0.05). Compared with the TGF-β1 group, the protein expression levels of fibrosis-related factors alpha-smooth muscle actin (α-SMA) and vimentin, and the mRNA expression levels of collagen Ⅰ and α-SMA were lower in the TGF-β1 + CID group (all P<0.05). (2) Compared with sham group, the mRNA and protein expression levels of GPR55 in UUO group were higher (both P<0.05). The serum creatinine in the UUO+CID group was lower compared to the UUO group ( P<0.05). There was no statistically significant difference in blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and creatine kinase isoenzyme between UUO group and UUO+CID group (all P>0.05). Compared with the UUO group, the protein expression levels of renal fibrosis-related factors fibronectin, collagen Ⅰ and vimentin, and the mRNA expression levels of fibronectin, collagen Ⅰ, collagen Ⅲ and α-SMA were lower in the UUO+CID group (all P<0.05). The degree of renal tubular dilation and interstitial collagen fiber deposition in the UUO+CID group was significantly reduced compared to the UUO group (all P<0.05). Conclusions:CID16020046 can reduce serum creatinine in UUO mice, protect renal function, and simultaneously decrease the expression of fibrosis-related factors in renal fibroblasts and mouse kidney tissues, thereby alleviating renal fibrosis.