Objective To ev aluate clinical results of selective anterior corpectomy combined with segmental discectomv and fusion for multilevel cervical myelopathv andits related factors.Methods Twenty-five patients of multilevel cervical myelopathv with 3 or 4 consecutive segments were treated with selective anterior corpectomy combinedwith segmental discectomy and fusion.The clinical results were evaluated by the parameters including improvement of Japanese Orthopaedic Association (JOA)score postoperatively,fusion rate,and the degree of spinal canal decompression and range of motion of the cervical spine.Results JOA score were improved from 9.5±1.3 pre-operatively to 13.8±0.8(P＜0.01)6 months post operation and 1 3.6±0.9(P＜0.05)atthe final follow-up.Bony fusion was verified by X-ray and 3D-CT scan 1 2 months post operation and final follow-up.Moreover,the degree of decompression of the spinal canal was also remarkable.The range of motion of the cervical spine after operation decreased significantly(P＜0.05).Conclusion Selective anterior corpectomy combined with segmental discectomy and fusion was an alternative choice for the surgical treatment of multilevel cervical myelopathy.

The aim of this study was to characterize the N and E protein encoding genes of a new human coronavirus (HCoV-NL63) which was identified from one of the clinical specimens (BJ8081) collected from a 12 years-old patient with acute respiratory infection in Beijing. The complete N and E gene sequences of HCoV-NL63 were amplified from clinical sample by RT-PCR, then were cloned into the pCF-T and pUCm-T vectors respectively and sequenced. The complete sequences of N and E genes were submitted to GenBank by Sequin and compared with N and E genes of prototype HCoV-NL63 and the other coronaviruses published in GenBank. The secondary structure and the characteristics of sample BJ8081 N and E proteins were predicted by bioinformatics. It was indicated that the N and E genes amplified from sample BJ8081 were 1134 bp and 234 bp in length and the predicted proteins including 377 amino acids and 77 amino acids, respectively. The data suggested that the region of amino acids 78-85 within N protein probably was the conserved region for all coronaviruses identified so far including HCoV-NL63. The region of amino acids 15-37 for E protein was probably the transmembrane domain. In conclusion, the recombinant plasmids pCF-T-8081 N and pUCm-T-8081 E were successfully constructed and sequenced, and the data predicted by bioinformatics are helpful for the further analysis of HCoV-NL63.
Amino Acid Sequence ; Child ; China ; Computational Biology ; methods ; Coronavirus ; classification ; genetics ; metabolism ; Coronavirus Infections ; virology ; Humans ; Molecular Sequence Data ; Nucleocapsid Proteins ; chemistry ; genetics ; metabolism ; Phylogeny ; Protein Structure, Secondary ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism

AIM: To observe thickness and morphological changes of bulbar conjunctiva pre- and post epidemic hemorrhagic conjunctivitis ( EHC ) therapy by optical coherence tomography ( OCT) .
METHODS: Observed morphological changes and measured the bulbar conjunctiva thicknesses of 29 cases (36 eyes) of incipient (1-2d) EHC patients, who were received and treated by department of ophthalmology, the Putuo Affiliated Hospital of Shanghai University of Traditional Chinese Medicine from May 2013 to December 2013, by OCT. Then measured the thickness again on 7, 14d after the therapy.
RESULTS: Among 29 patients (36 eyes), 7d after the EHC therapy, in 27 cured eyes, the full-thickness ( before 344. 00±59. 91μm, after 230. 19±22. 16μm, t=11. 75, P<0-01); epithelial thickness ( before 56. 52±6. 19μm, after 51. 37±5. 53μm, t=4. 61, P<0. 01); and stromal thickness (before 287. 11±60. 56μm, after 178. 81±20. 20μm, t=10. 69, P<0. 01) of patients' bulbar conjunctiva were thicker than values measured after therapy with significant difference. Significant difference was also found for full-thickness ( before 361. 39±65. 56μm, after 233. 44±22. 57μm, the difference was statistically significant, t=14. 45, P<0. 01);epithelial thickness ( before 55. 50±6. 72μm, after 46. 67±5-24μm, t=10. 06, P<0. 01) and stromal thickness ( before 305. 61±66. 02μm, after 186. 78±21. 82μm, t=13. 11, P<0-01 ) of patients' bulbar conjunctiva between values measured before and 14d after therapy.
CONCLUSION: The OCT is able to measure the thickness of bulbar conjunctiva in EHC patients. An significant increase was found in full, epithelial and stromal thickness of EHC patients' bulbar conjunctiva. With recovery from the disease, subepithelial fluid, interlaminar fluid and edema of the bulbar conjunctival stroma faded away firstly, which provide references for clinical therapies of the EHC.

Objective To find out the distribution pattern of environmental fluoride in the coal-burning endemic fluorosis areas to provide scientific evidence for establishing prevention and remedial measures in Chongqing. Methods According to historical data in Chongqing In 2008, 4 endemic villages(Lingyun and Lizi Villages of Wushan County, and Taiping and Daqing Villages of Pengshui County) and 2 non-endemic villages (ShuangLou and XianLong Villages of Yongchuan County) were investigated. Dental fluorosis of children in 8 to 12 year old and residents over 16 years of age for clinical skeletal fluorosis were examined in four endemic villages. Five households from each of 6 villages were taken, where 500 g of coal, mixed clay with coal, mixed coal of soil, coal cinder, soil were sampled; 15 people were taken in each village, each household gathering 500 g of corn, rice, potatoes, vegetables, grain and vegetable, 100 g of dried pepper and 250 ml of drinking water were sampled from 15 families of each village. Household drinking water samples were collected 1, each 250 ml. For those having tea-drinking habit, each household was collected 50 g of tea and 600 ml of drinking tea, the amount of fluoride were determined. Indoor and outdoor air was collected and measured in 5 households in each village. Results The detected rate of dental fluorosis of children in endemic areas was 74.65% (736/986). The detected rate of skeletal fluorosis of adult was 7.20%(736/986). The average fluoride content of coal, mixed clay with coal, mixed coal of soil, cinder coal, soil in the endemic villages was (310.56±209.46), (360.51±224.96), (293.62±65.15), (186.59±133.66), (497.54±294.70)mg/kg. The average fluoride content in non-endemic villages was (48.68±10.62), (275.66±62.69), (152.20±34.43), (209.14±188.66),269.98±58.21)mg/kg. The fluoride content level of endemic villages was significantly higher than that of non-endemic villages(t=7.67,31.54,5.82, 5.82, all P<0.05). The average fluoride content of drinking water, corn, pepper, flee, potato and vegetable in the endemic villages was (0.30±0.14)mg/L, (1.83±2.67), (23.50±91.80), (0.77±0.25), (0.44±0.11), (0.48±0.18)mg/kg, The average fluoride content in non-endemic village was (0.18±0.06)mg/L, (2.21±0.46), (2.82±2.51), (1.31±0.21), (0.64±0.41), (1.10±0.77)mg/kg. The fluoride content in drinking water and pepper in the endemic villages was significantly higher than that of the non-endemic villages(t=7.79, 2.33, all P<0.05). The fluoride content of rice, potato and vegetable in the non-endemic villages was significantly higher than that of the endemic villages(t=39.29,4.69,4.01, all P<0.05). There was no significant difference of fluoride content of tea and drinking tea between endemic villages[(99.41±55.83)mg/kg, (1.59±0.91)mg/L] and non-endemic villages[(79.95±43.78)mg/kg, (1.80±1.16)mg/L, t=1.01, 0.27, all P>0.05]. The amount of drinking tea in the endemic village[(1.45±0.68)L/d] was higher than that in non-endemic village[(1.00±0.47)L/d, t=4.27, P<0.05]. The average fluoride content of indoor air in the endemic village[(12.77±8.08)μg/m3] was higher than that in non-endemic village [(1.16±1.08)μg/m3, t=9.49, P<0.01]. There was no significant difference of fluoride content of outdoor air between endemic village and non-endemic village[(1.10±1.57), (0.39±0.31)μg/m3, t=2.01, P>0.05)]. Conclusions The fluoride source of coal-burning endemic fluorosis areas are coal and mixed coal of soil in Chongqing. Fluoride enters into human bodies mainly via respiratory, not from food. Although fluoride is rich in pepper, people don't eat it, so reducing the fluoride content in indoor air is the principle measure. Drinking tea may be was one factor of endemic fluorosis, which needs to be further studied.

Objective To analyze the spatio-temporal process on 2009 influenza A (HlNl) pandemic in Changsha and the influencing factors during the diffusion process. Methods Data were from the following 5 sources, influenza A (HlNl) pandemic gathered in 2009, Geographic Information System (GIS) of Changsha, the broad range of theorems and techniques of hot spot analysis, spatio-temporal process analysis and Spearman correlation analysis. Results Hot spot areas appeared to be more in the economically developed areas, such as cities and townships. The cluster of spatial-temporal distribution of influenza A (HlNl) pandemic was most likely appearing in Liuyang city (RR=22.70,P<0.01). The secondary cluster would include districts as Yuelu (RR=6A9,P< 0.01) , Yuhua (RR=81.63, P<0.01). Xingsha township appeared as the center in the Changsha county (RR=2.90, P<0.01) while townships as Yutangping (RR=19.31, P<0.01) , Chengjiao (RR=73.14,P<0.01) and Longtian appeared as the center in the west of Ningxiang county (RR= 14.43,P<0.01) and Wushan as the center in the Wangcheng county (RR= 13.84,P<0.01). As time went on, the epidemic moved towards the eastern and more developed regions. Regarding factor analysis, population, the amount of students, geographic relationship and business activities etc. appeared to be the key elements influencing the transmission of influenza A (H1N1) pandemic. At the beginning of the epidemic, population density served as the main factor (r=0.477, P<0.05) but during the initial and fast growing stages, it was replaced by the size of students to serve as the important indicator (r=0.831, P<0.01; r=0.518, P<0.01). However, during the peak of the epidemics, the business activities played an important role (r=-0.676, P<0.01). Conclusion Groups under high risk and districts with high incidence rates were shifting, along with the temporal process of influenza A(H1N1) pandemic, suggesting that the protection measures need to be adjusted, according to the significance of influencing factors at different stages.

Ocular neovascularization is a pathological change in various ocular diseases such as diabetic retinopathy, retinopathy of prematurity, central retinal vein occlusion and age-related macular degeneration, which seriously affects patient's vision. β receptors are expressed in conjunctiva, corneal epithelial cells, corneal endothelial cells, extraocular muscles, trabecular meshwork, ciliary muscle, lens and retina. β adrenergic receptor antagonists bind to β receptors to exert anti-angiogenic effects by inhibiting the expression of vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1, interleukin-6 and other angiogenic cytokines; reducing macrophage-related inflammatory response; increasing the expression of anti-angiogenic factors. In the treatment of corneal neovascularization, choroidal neovascularization, and retinopathy of prematurity, it can significantly reduce the area of neovascularization and delay disease progression. Co-administration of anti-VEGF drugs can reduce the frequency of administration of anti-VEGF drugs. At effective therapeutic concentrations, β-adrenergic receptor antagonists are well tolerated; they have broader targets than anti-VEGF drugs, which offers new treatment strategies for ocular neovascularization such as corneal, choroidal and retinal neovascularization.

Objective:To explore the effects of apolipoprotein E-mimetic peptide COG1410 on M1/M2 microglia polarization and retinal ganglion cells (RGCs) survival after ischemia-reperfusion (IR) injury in the mouse retina and its possible mechanisms.Methods:Eighteen 8-week-old C57BL/6J male mice were divided into control group (6 mice), IR 3 days group (6 mice), IR 7 days group (3 mice), and IR 14 days group (3 mice) according to the randomized number table method.Mice in IR group were perfused in the anterior chamber using saline, and the intraocular pressure (IOP) was raised to 100 mmHg (1 mmHg=0.133 kPa) and maintained for 1 hour in order to establish a model of IR injury in the retina.Three mice from control group and 3 mice from IR 3 days group were taken to observe the distribution of retinal microglia by immunofluorescence staining of retinal frozen sections.Three mice were taken from normal control, IR 3 days, IR 7 days, and IR 14 days groups respectively to observe the changes of retinal M1-type and M2-type microglial cells with time after IR injury by immunofluorescence staining of retina.Another 91 C57BL/6J mice were randomly divided into normal control group (19 mice), IR group (24 mice), saline group (24 mice), and COG1410 group (24 mice) according to the random number table method.Mice in normal control group maintained a normal IOP, and the IR injury model was established in the other three groups.In addition, COG1410 group and saline group were injected with 1 mg/kg COG1410 and an equal volume of saline by tail vein injection, respectively.The microglia phenotype and survival rate of RGCs were observed by immunofluorescence staining of retinal wholemount.The relative expressions of retinal tumor necrosis factor-ɑ (TNF-ɑ) and interleukin-1β (IL-1β) mRNA were detected by real-time fluorescence quantitative PCR.The apoptosis of retinal neuronal cells was observed by the TUNEL assay.The expression levels of retinal nuclear factor-κB (NF-κB), B lymphocyte-2 (Bcl-2), and Bcl-2-associated X protein (Bax) proteins were detected by Western blot.Use and care of animals strictly complied with the Hubei Provincial Regulations on the Management of Laboratory Animals and the experiment was approved by the Animal Ethics Committee of the Renmin Hospital of Wuhan University (No.WDRM20190113).Results:Retinal microglia in normal control group and IR 3 days group were mainly distributed in the ganglion cell layer, inner plexiform layer, and outer plexiform layer.There were statistically significant differences in the comparison of the proportions of M1-type and M2-type microglia among normal control, IR 3 days, IR 7 days, and IR 14 days groups ( F=29.83, 57.62; both at P<0.001). Compared with normal control group, the number of M1-type microglia was higher in IR 3 days group, and the number of M2-type microglia was higher in IR 7 days group, and the differences were statistically significant (all at P<0.05). The proportions of M1-type microglia in normal control group, IR group, saline group, and COG1410 group were (4.25±0.57)%, (65.26±10.43)%, (63.01±4.93)%, and (33.13±4.46%), respectively, and the proportions of M2-type microglia in the four groups were (4.50±0.20)%, (11.47±0.24 )%, (11.75±0.17)%, and (38.93±4.26)%, showing statistically significant differences among them ( F=23.33, 50.82; both at P<0.001). The proportions of M1-type microglia decreased while the proportions of M2-type microglia increased in COG1410 group when compared with IR group, and the differences were statistically significant (both at P<0.05). There were statistically significant differences in RGCs survival rate, relative expression of retinal TNF-ɑ and IL-1β mRNA, retinal apoptotic cell count, retinal NF-κB and Bax protein expression levels, and Bax/Bcl-2 ratio among the four groups ( F=30.77, 12.52, 6.74, 28.72, 13.02, 7.94, 7.58; all at P<0.05). Compared with normal control group, there were significant decreases in the survival rate of RGCs and increases in retinal apoptotic cell number, TNF-ɑ and IL-1β mRNA expression, retinal NF-κB and Bax protein expression levels, and Bax/Bcl-2 ratio in IR group (all at P<0.05). Compared with IR group, the COG1410 group had increased retinal RGCs survival rate, decreased TNF-ɑ and IL-1β mRNA expression levels, decreased TUNEL-positive cells, decreased NF-κB and Bax proteins expression levels, and decreased Bax/Bcl2 ratio, and the differences were statistically significant (all at P<0.05). Conclusions:Three days after retinal IR modeling, COG1410 promotes the polarization of M1-type microglia to M2-type, inhibits the expression of retinal NF-κB and downstream inflammatory factors, and attenuates the retinal inflammatory response, as well as inhibits the expression of apoptosis-related proteins, which promotes the survival of RGCs.

This study was purposed to evaluate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on hematopoietic reconstruction and survival in beagles exposed to mixed fission neutron and γ-ray. 13 beagles were unilaterally exposed to single dose of 2.3 Gy 90% neutrons. The experiments were divided into 3 groups: irradiation control group (no any treatment, n = 4), supportive care group (n = 5) and rhG-CSF plus supportive care group (n = 4, abbreviated as rhG-CSF group) in which the beagles were subcutaneously injected with 200 µg/kg of rhG-CSF early at half an hour and 24 hours post-irradiation respectively. The results showed that 2.3 Gy 90% neutron irradiation induced a severe acute radiation sickness of bone marrow type. The administration of rhG-CSF increased the survival rate from 60% in supportive care group to 100%. Twice injection of rhG-CSF in the first 24 hours reduced duration of neutropenia, enhanced neutrophil nadir and promoted neutrophil recovery when compared with control cohort administered clinical support. The number of colony-forming cells (CFU-GM, CFU-E, and BFU-E) in peripheral blood of rhG-CSF treated canines increased 2-to 5-fold relative to those of the supportive care group on day 3. All canines treated with rhG-CSF achieved hematopoietic reconstruction as evidenced by the pathological section of sternum while severe shortage of hemopoietic cells remained in the cohorts given supportive care alone. It is concluded that the combination of supportive care and high-dose rhG-CSF can accelerate hematopoietic recovery and enhance survival of dogs exposed to 2.3 Gy mixed neutron and gamma ray.
Animals ; Dogs ; Gamma Rays ; adverse effects ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic System ; drug effects ; radiation effects ; Neutron Diffraction ; Recombinant Proteins ; administration & dosage ; pharmacology ; Survival Rate

Leigh syndrome is a genetically heterogeneous disease caused by defects in enzymes involved in aerobic energy metabolism and the Krebs' cycle. Deficiency of pyruvate dehydrogenase complex E1 alpha subunit (PDHA1) is the common cause of Leigh syndrome. In this study, one Chinese case of PDHA1 deficiency was reported. The patient was a boy with normal mental development, retarded motor development, general weakness, hypotonia and areflexia. Muscle histopathological findings suggested axonal peripheral neuropathy. Brain magnetic resonance imaging at 5 years of age revealed bilateral putamina lesions and periventricular white matter demyelination, supporting the diagnosis of Leigh syndrome. A C214T mutation in exon 3 of the PDHA1 gene was detected. After the treatment of thiamin, coenzyme Q10, Lcarnitine and carbohydrates-restricted diet, his movement ability improved significantly. At present, the patient is 8 years old and has normal school life. PDHA1 deficiency is an X-linked inherited metabolic disease, which shares various clinical manifestations and leads to difficult diagnosis. This patient predominately presented with progressive weakness and was diagnosed by gene analysis.
Child, Preschool ; Diagnosis, Differential ; Humans ; Leigh Disease ; diagnosis ; genetics ; therapy ; Male ; Mutation ; Pyruvate Dehydrogenase (Lipoamide) ; genetics

OBJECTIVETo reveal the etiological agent of hand, foot and mouth disease in children in Beijing.

METHODSThroat swabs were collected from 6 infants and young children with hand, foot and mouth disease who visited the affiliated Children's Hospital from May to June 2007. Aspirated fluid from tracheal intubatton, serum and cerebral spinal fluid (CSF) were collected from a 9 years old girl (No.4243) having central neural system complication of severe hand, foot and mouth disease and admitted to the hospital from the Emergency Department. Throat swab and aspirated fluid were inoculated into the cell lines Hep-2, MDCK and Vero for virus isolation. RNAs were extracted by Trizol from 6 throat swab specimens and aspirated fluid, serum while CSF was from that severe case (No.4243). The gene fragment from 5' UTR of enterovirus was amplified from throat swabs and aspirated fluid by reverse transcription-polymerase chain reaction (RT-PCR) with the primer pairs located at the untranslated region of all enterovirus. EV71 was identified by RT-PCR with the 2 and half primer pairs located at different parts of VP1 gene of EV71. The PCR products for VP1 encoding gene of EV71 from the specimens were sequenced and sequence analysis was performed by comparing those published VP1 genes of EV71. EV71 and CA16 specific primers were used to identify the isolates by RT-PCR and the sequences were directly determined from PCR products.

RESULTSGene fragments with expected molecular weight were amplified from all 6 throat swabs and the aspirate by the primer pairs universal for the 5' UTR of enterovirus, suggesting that these patients with hand, foot and mouth disease were infected by entorovirus. Out of these 7 specimens, 2 throat swabs and the aspirate were also showing positive for the VP1 of the EV71 by different primer sets. Sequence analysis revealed that the sequences for the amplicons from 1 throat swab (No. F4211) and the aspirate shared highest homology with those published EV71, indicating that these specimens were truly positive for EV71. The sequences amplified from these specimens shared 100% and 98.9% homology to each other and were closer to the sequences of EV71 identified from Zhejiang province than those from Taiwan and strain BrCr. Gene fragments for 5' UTR of enterovirus were obtained by RT-PCR after CPE appeared in 6 out of 7 inoculations including that aspirate fluid in Vero cell, indicating that enteroviruses were isolated from these specimens. Virus isolates from one throat swab (No. F4211) and the aspirate (No. 4243) were positive by RT-PCR with the primer pairs for EV71, which was consistent with RT-PCR amplification directly from specimens. Virus isolates from other 4 specimens were CA16 by RT-PCR and sequence analysis.

CONCLUSIONThese data suggested that hand, foot and mouth disease recently appeared in children in Beijing was related with EV71 and CA16. EV71 could cause severe clinical manifestations with central nerve system complications even in the child older than 5 years.

Animals ; Cercopithecus aethiops ; Child ; China ; epidemiology ; Enterovirus A, Human ; genetics ; isolation & purification ; Enterovirus Infections ; complications ; epidemiology ; Female ; Hand, Foot and Mouth Disease ; etiology ; virology ; Humans ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, RNA ; Vero Cells ; Virus Cultivation