Vitreoscilla Hemoglobin(VHb), with the function of increasing the growth of and product yield by a heterologous host, has been widely use in the area of fermentation, environment protection, transgenic animal and plant, recombinant protein expression, etc. Fusion protein of VHb with other enzyme or protein can enhance activity and stability of the enzyme or isolation efficiency of the protein. The reconstitution of VHb will be helpful to obtain ‘novel’ proteins which have better activity.

Background Compare with the lamellar laser refractive surgery,the laser slip on the surface operation is a better method in the correct of refractive error without the flap-related complications.However,whether to remain corneal epithelial flap and its influence for epithelial healing and visual restoration are still under investigation.Objective The aim of this study was to compare the short-term clinical efficacy of epipolis laser insitu keratomileusis (Epi-LASIK) and flap-free Epi-LASIK for correction of refractive error in high myopia.Methods The paired control design was adopted.Sixty eyes of consecutive 30 patients with high myopia were collected in this study.The written consent form was obtained from each patients prior to operation.Regular EpiLASIK was performed in the right eyes and the flap-free Epi-LASIK was carried out in the left eyes.Patients were followed-up for one month.The postoperative ocular irritation symptoms were scored,and the healing period of corneal epithelium and uncorrected visual acuity were examined at 1,3 and 5 days after operation.The naked visual acuity,diopter,best corrected visual acuity and the grading of haze were recorded and compared between the two groups at one month after operation.Results After surgery,the scores of corneal irritation were 3.13 ±0.43 and 3.30±0.47on the first day,showing insignificant differences between these two groups (t =- 1.54,P＞0.05 ).However,there existed statistically significant difference in the scores of corneal irritation on the third day after operation between the two groups (t =- 17.95,P＜0.01 ).The average time of corneal epithelial healing were (2.58 ±0.49 ) days in the flapfree Epi-LASIK group and ( 3.98 ±0.74) days in Epi-LASIK group with the significant difference between them (t=-11.36,P＜0.01 ).There was no statistical difference in uncorrected visual acuity in the first day (0.59±0.12 vs 0.58±0.11,t=0.39,P＞0.05),the fifth day (0.69±0.08 vs 0.67±0.09,t=0.84,P＞0.05) and 1 month (t =-1.90,P＞0.05 ).No significant difference was seen in the diopter between the two groups in 1 month after surgery (-1.03±0.45 vs -1.07 ±0.38,t =-0.31,P＞0.05 ).The eye numbers of corneal haze for 0 grade were 27 ( 90.00％ ) and over the 0.5 grade were 3 ( 10.00％ ) in flap-free Epi-LASIK group respectively,and those of corneal haze for 0 grade were 26(86.66％ ) and above the 0.5 grade were 4 (13.33％) in Epi-LASIK group respectively,showing a considerably difference between the two groups (x2 =0.0031,P＞0.05).Conclusions Both the flapfree or reserved epithelial Epi-LASIK allow the safe and effective correction of refractive error in high myopia,showing good predictability and stability in short term.Flap-free Epi-LASIK can relieve the corneal irritation symptom and accelerate the healing of corneal epithelium and recovery of visual acuity.

Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual funetion.However,the color vision system of animals which encounter uncertainty of spectral signals should be plastic.Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear.Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light.Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed tO the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively.The flat-mounted retinal sample was prepared and divided into dorsal zone,ventral zone and mixed zone anatomically according to the distribution of difierent light-sensitive cone.The changes in density of cone cells sensitivited to different colored light were detected by single-1abel or double-label immunocytochemistry.The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively.Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group,purple light group and white light group (F=234.28,P<0.01).Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group wag evidently lower(q=389.68,P<0.01；q=67.11,P<0.01).No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group,purple light group and white light group(F=3.14,P>0.05).The density of coexpression of the mixed cone cells was increased in green light group(q=157.55,P<0.01)but decreased in purple light roup (q=254.85,P<0.01)in comparison with white light group.The expression levels of green-opsin and green-opsin mRNA in green light group was significantly elevated(q=184.45,P<0.01；q=4.71,P<0.05),but those of purple light group were evidently declined(q=5.87,P<0.05；q=346.66,P<0.01)in comparison with white light group.There was no statistically significant differences were found in the expression of purple-opsin and its mRNA among all the groups(F=1.24,P>0.05；F=3.27,P>0.05).Conclusion After the exposure of long-time monochromatic light illumination,monochromatic cones density and its opsin in guinea pig occur the corresponding alteration to gain good spatial vision as a compensatory reaction.These outcomes imply that there is some plasticity during the development of color vision.The increase of green-sensitive cones might be from the differentiation of coexpression cones in transition region.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis and calcium channels of PC12 cells induced by ischemic and hypoxia and its mechanisms. Methods PC12 cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing HSP70 and green fluorescent protein (GFP) fluorescin gene], lentivirus control group (infected by lentivirus containing GFP without HSP70 gene) and non-infection group. PC12 cells were subjected ischemia/hypoxia for 4, 8, 12, 24 hours, and the cell activity was determined by methylthiazolyl tetrazolium (MTT) assay test inorder to determine the best time for ischemia/hypoxia. The mRNA expressions of HSP70, muscle/endoplasmic reticulum Ca2+-ATP isoforms (SERCA2a, SERCA2b), ryanodine receptor 2 (RyR2), and inositol 1,4,5-triphosphate receptor 1 (IP3R1) were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expressions of HSP70, SERCA, and IP3R were determined by Western Blot at 8 hours after ischemic/hypoxia. Flow cytometry was used to determine the levels of intracellular reactive oxygen (ROS) and intracellular Ca2+ ([Ca2+]i). Results With the prolongation of time of ischemia/hypoxia, the cell viability in all groups showed an increase followed by a weakening, and peaked at 8 hours. The cell viability at 8 hours in lentiviral infection group was significantly higher than that of the non-infection group and lentivirus control group [A value (×10-2): 20.3±2.2 vs. 14.1±2.1, 15.0±1.6, both P < 0.01], the mRNA and protein expressions of HSP70 and SERCA in lentiviral infection group were significantly increased [HSP70 mRNA (2-ΔΔCt ): 0.785±0.018 vs. 0.428±0.019, 0.423±0.023; HSP70 protein (gray value): 2.72±0.20 vs. 1.56±0.36, 1.63±0.41; SERCA2a mRNA (2-ΔΔCt ): 0.971±0.037 vs. 0.367±0.014, 0.347±0.012; SERCA2b mRNA (2-ΔΔCt ): 8.869±0.162 vs. 3.015±0.091, 2.941±0.091; SERCA protein (gray value): 2.84±0.18 vs. 1.48±0.26, 1.52±0.29], and IP3R2 mRNA and protein expressions were significantly declined [IP3R2 mRNA (2-ΔΔCt ): 0.183±0.020 vs. 0.439±0.020, 0.433±0.040; IP3R2 protein (gray value): 1.15±0.12 vs. 1.91±0.20, 1.83±0.19], with statistically significant differences (all P < 0.01); no significant difference in RyR mRNA was found [2-ΔΔCt (×10-3): 1.97±0.63 vs. 2.02±0.22, 2.01±0.09, both P > 0.05]; the relative fluorescence intensity of ROS and [Ca2+]i in lentiviral infection group was significantly reduced (ROS: 30.54±1.23 vs. 58.03±1.97, 57.72±2.35; [Ca2+]i: 34.50±2.05 vs. 48.20±3.02, 46.80±2.75, all P < 0.01]. Conclusion Exogenous HSP70 can maintain calcium homeostasis in the endoplasmic reticulum of PC12 cells, affect the Ca2+ channel protein regulated by calcium channel IP3R and calcium pump SERCA, which may cause hypoxia/ischemia intracellular injury.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

Objectives: Blood and urine are commonly used specimens for clinical testing, and their contents, particularly exosomal microRNA (miRNA), are diverse, reflecting the metabolic activities of tissues and organs in the body. Methods: Blood and urine samples were collected from six healthy adults. Exosomes were then enriched from these samples, followed by sequencing and bioinformatic analysis of exosomal miRNA. Results: The comparative analysis of miRNAs in blood and urine revealed that 41 miRNAs were more abundant in blood, while 61 were found at lower levels. Notably, hsa-miR-934 was among those with higher expression in blood, whereas hsa-miR-425-5p was one of the miRNAs with lower expression.Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that the target mRNAs of differentially expressed exosomal miRNAs (DEexo-miRNAs) in both blood and urine are implicated in various signaling pathways, including proteoglycans in cancer, axonal guidance, and the regulation of the actin cytoskeleton. Additionally, the target mRNAs associated with DEexo-miRNAs in urine were also linked to processes such as ubiquitin-mediated proteolysis and the phosphatidylinositol signaling system. In contrast, the target mRNAs corresponding to DEexo-miRNAs in blood were involved in the FoxO signaling pathway and chronic myeloid leukemia, among others. Conclusion This study observed differential expression of exosomal miRNAs in blood and urine, thereby enriching the available library of exosomal miRNA for these two sample types. It also lays the groundwork for the detection of exosomal biomarkers from blood and urine.

ObjectiveTo investigate the effect of rehabilitative treatment on different groups of infants with cerebral palsy in different age, sickness extent and rehabilitative period.MethodsBy the prospective method, 90 patients from 3 to 36 months old were randomly divided into three groups including 3 to 12 months, 13 to 24 months and 25 to 36 months with 30 cases each. Every 10 cases in light, moderate and severe degrees were given rehabilitative care for 2 months and 3 months respectively. The personal ADL of every case was evaluated at the initial and the final state. The compared score was analyzed with T test.ResultsComparing the groups of different ages, there were prominent differences of treatment under the same sickness, among the light, moderate and severe degrees(P<0.01). The rehabilitative treatment effect of 3 months was better than that of 2 months. Furthermore the effect of the group of 3-12 months was better than that of 25-36 months.ConclusionsThe rehabilitative care is prominently effective in the light and moderate groups , the former is better and within 3 months is better than 2 months. The younger infant is, the better the rehabilitative treatment is.

Objective - ，，，，，- - - To establish a gas chromatographic method for the determination of trans 1 1 1 4 4 4 hexafluoro 2 ［ - （）］ Methods - （） butene HFO 1366mzz E in workplace air. HFO 1366mzz E in air was directly collected with aluminum foil ， ， ， composite plastic bag separated by dimethylpolysiloxane capillary column detected by flame ionization detector and Results - （） - 3， quantified with external standard method. The linear range of HFO 1366mzz E was 6.82 68 200.00 mg/m with the 3， correlation coefficient of 0.999 9. The detection limit and the lower limit of quantitation were 0.59 and 1.98 mg/m respectively. - - - The recovery rate was within 95.45% 103.05%. The relative standard deviation of within batch precision and between batch - - ， precision were 2.26% 5.07% and 4.09% 6.82% respectively. The samples can be stored at room temperature for at least seven Conclusion ， ， days. This method is simple to use with a wide linear range low detection limit high accuracy and precision and - （） good sample stability. It can be used for the detection of HFO 1366mzz E in the air of workplace

OBJECTIVETo observe the clinical efficiency of curettage combining circumcision on giant condyloma acuminata(CA) in male external genitalia, the relationship between recurrence and curettage depth, the possibility of HPV infection in PBMC after operation.

METHODSCurettage combining circumcision was carried out on 50 cases with CA. The removed wart and wound surface tissues were examined under light microcope and for HPV-DNA detection by PCR. HPV-DNA was detected in PBMC during < or =1 and > or =2 weeks after operation.

RESULTS(1) 46 cases were cured completely after one treating (once the cure rate is 92%), 4 cases twice(twice the cure rate is 8%); (2) The tissues were proved to be HPV-DNA positive by PCR amplification and CA relapse occurred; (3) HPV-DNA was detected in PBMC only within the week after curettage in the 19 cases.

CONCLUSIONS(1) The clinical efficiency of curettage combining circumcision on giant condyloma acuminata in male external genitalia is sure; (2) Cure rate and relapse rate are related with curettage depth; (3) Transient positive HPV-DNA in PBMC may be detected.

Adolescent ; Adult ; Aged ; Circumcision, Male ; Condylomata Acuminata ; surgery ; Curettage ; methods ; Humans ; Male ; Middle Aged ; Young Adult

Objective To analyze the disease burden caused by occupational coal workers' pneumoconiosis (CWP) in Anhui Methods Province. A total of 1 649 patients with occupational CWP diagnosed in Anhui Province from 1950 to 2019 were selected as the study subjects using a retrospective study method. Age, gender, survival time, location, working age of disease onset, age of death, stage and date of diagnosis of CWP, comorbidities at the time of investigation, hospitalization and outpatient expenses in the past year, cost of absence from work, cost of absence from work of caregivers, and cost of nutrition and transportation were investigated. The years of life lost, years lived with disability (YLDs), disability adjusted life years (DALYs) Results and economic losses were calculated. Among these patients, 1 405 cases survived and 244 cases died. In the age range - - - of 20.0 70.0 years, the YLDs of surviving patients were 2.12 22.20 (8.19±3.59) person years. The YLDs of patients with disease vs - P complications was higher than that of patients without complications [(8.55±3.95) (7.87±3.21) person years, <0.01]. The total - - DALYs of the patients was 14 031.59 person years, and the average per capita DALYs was 8.51 person years. Its YLDs accounted for 82.0 % of the total DALYs. The total economic loss caused by CWP in the 1 405 surviving patients was 354.903 0 Conclusion million yuan, and the average per capita economic loss was 252 600 yuan. The disease burden caused by CWP is relatively high in Anhui Province. In addition to early detection, diagnosis and treatment, it is necessary to focus on prevention and treatment of CWP complications to reduce the disease burden.