Establishment and validation of embryo high-quality prediction models based on the third-day 340 nm absorbance embryo culture / 中国组织工程研究

BACKGROUND:A large number of previous studies have confirmed that a high concentration of metabolites is significantly correlated with embryo quality and clinical outcome,and the theory of silencing embryo development indicates that normally developed embryos maintain a low level of material exchange with the outside world during in vitro culture,while embryos often show abnormal metabolic activity due to stress repair mechanism when DNA damage occurs. OBJECTIVE:To establish and verify an embryo quality prediction model based on the third-day 340 nm absorbance embryo cultures to provide the basis for a more objective and accurate embryo quality assessment. METHODS:269 patients at the Nanxishan Hospital of Guangxi Zhuang Autonomous Region for in vitro fertilization and embryo transplantation from November 2019 to December 2021 were retrospectively analyzed.Among them,on day 3,162 cases who had 873 optimal embryos and 214 high-quality blastocysts were included in the high-quality embryo group.On day 3,107 cases who had 859 non-optimal embryos and 214 non-high-quality blastocysts were included in the non-high-quality embryo group.Lambert-beer law was used to screen out the characteristic wavelength with distinguishing degree between superior and non-superior embryos,analyze its correlation and influence trend with high-quality embryos,and establish the clinical prediction model and validation of absorbance for high-quality and non-high-quality embryos at this wavelength. RESULTS AND CONCLUSION:(1)There was a significant difference in absorbance between high-quality and non-high-quality embryos at 340 nm on day 3(P<0.001),and a negative correlation was found with the formation of high-quality embryos on day 3(r=-0.486,P<0.001).The absorbance of high-quality and non-high-quality blastocyst at 340 nm was significantly different(P<0.05),and was negatively correlated with the formation of high-quality blastocyst(r=-0.642,P<0.001).(2)The optimal cut-off value of absorbance at 340 nm between high-quality and non-high-quality embryos on day 3 was 0.235.The area under the curve was 0.799.Sensitivity was 62.9%.Specificity was 78.0%.Accuracy was 70.5%.The optimum cutoff value of high-quality and non-high-quality blastocysts of absorbance at 340 nm was 0.175.The area under the curve was 0.871.Sensitivity was 74.3%.Specificity was 89.1%.Accuracy was 82.2%.(3)Restricted cubic spline curve analysis showed that when the absorbance of the culture medium at 340 nm was greater than 0.221,there was a significant positive trend on the formation of non-high-quality embryos at day 3,and when the absorbance of the culture medium at 340 nm was greater than 0.160,there was a significant positive trend on the formation of non-high-quality blastocysts.(4)The clinical decision curve and clinical influence curve showed that the absorbance of the culture medium at 340 nm had the maximum clinical net benefit for the prediction models of high-quality embryos and high-quality blastocysts on the third day when the valve probability was 0.18-0.95 and 0.16-1.00,respectively,and the ratio of loss to gain within the valve probability range was always less than 1.It is proven that the prediction model has good efficacy in clinical applications.The results of embryo transfer showed that the absorbance of embryo culture medium at 340 nm in non-pregnant patients was significantly higher than that in clinical pregnancy,biochemical pregnancy and early abortion patients(P<0.05).(5)The high-quality and non-high-quality embryo culture in 340 nm absorbance has a significant difference with correlation.The embryo quality prediction model has a certain clinical value and application effectiveness.The joint embryo morphology evaluation to a certain extent improves the objectivity and accuracy of embryo quality evaluation.

Construction and validation of a nomogram model to predict abnormal female factors in in vitro fertilization / 中国组织工程研究

BACKGROUND:Reducing the rate of abnormal fertilization is an effective approach to improving the efficacy of in vitro fertilization and reducing patients'financial strain.However,the current research on abnormal fertilization has focused on exploring the types of prokaryotic nuclei and their generation mechanisms,as well as analyzing embryos formed by abnormal fertilization,chromosomal ploidy and utilization value.There is a lack of clinical prediction models for abnormal fertilization based on retrospective studies. OBJECTIVE:To construct a nomogram model to predict abnormal female factors in in vitro fertilization. METHODS:A total of 5 075 patients undergoing treatment for conventional in vitro fertilization at Nanxishan Hospital of Guangxi Zhuang Autonomous Region from March 2017 to March 2022 were retrospectively analyzed.The male confounders were calibrated on a 1:1 propensity score with a match tolerance of 0.02,and 1 672 cases were successfully matched.According to the Vienna Consensus,patients with≥60%normal fertilization capacity were included in the normal fertilization group(n=836)and those with<60%normal fertilization capacity were included in the abnormal fertilization group(n=836).The model and validation groups were obtained by random sampling at a ratio of 7:3.Factors related to the occurrence of abnormal fertilization following conventional in vitro fertilization in the model group were screened using univariate analysis and the best matching factors were selected using the Least Absolute Shrinkage and Selection Operator(LASSO)and included in a multifactorial forward stepwise Logistic regression to identify their independent influencing factors and plot a nomogram.Finally,the prediction model was validated for discrimination,accuracy and clinical application efficacy using receiver operating characteristic curves,calibration curves,clinical decision curves and clinical impact curves. RESULTS AND CONCLUSION:The univariate analysis indicated the factors influencing the occurrence of abnormal fertilization were age,controlled ovarian hyperstimulation protocol,number of assisted pregnancies,years of infertility,infertility factors,anti-mullerian hormone,sinus follicle count,basal luteinizing hormone,luteinizing hormone concentration on the human chorionic gonadotropin day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).LASSO regression further identified the best matching factors,including age,microstimulation protocol,number of assisted pregnancies,years of infertility,anti-mullerian hormone,luteinizing hormone level on human chorionic gonadotropin injection day,and estradiol level on human chorionic gonadotropin injection day(P<0.05).Multifactorial forward stepwise Logistic regression results showed that age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day were independent influencing factors for the occurrence of abnormal fertilization following conventional in vitro fertilization.The receiver operating characteristic curves showed an area under the curve of 0.761(0.746,0.777)for the model group and 0.767(0.733,0.801)for the validation group,indicating that the model has good discrimination.The mean absolute error of the calibration curve was 0.044,and the Hosmer-Lemeshow test indicated that there was no significant difference between the predicted probability of abnormal fertilization and the actual probability of abnormal fertilization(P>0.05),indicating the prediction model has good consistency and accuracy.The clinical decision curves and clinical impact curves showed that the model and validation groups had the maximum net clinical benefit at valve probability values of 0.00-0.52 and 0.00-0.48,respectively,and there was a good clinical application efficacy in this valve probability range.To conclude,the nomogram model has good discrimination and accuracy as well as clinical application efficacy for predicting the occurrence of abnormal fertilization in women undergoing conventional in vitro fertilization based on age,microstimulation protocol,number of assisted conceptions,years of infertility,anti-mullerian hormone,and estradiol level on human chorionic gonadotropin injection day.

Construction of nomogram and validation of clinical prediction model for high-quality blastocyst formation in patients with unexplained infertility / 中国组织工程研究

BACKGROUND:Unexplained infertility is associated with a higher abortion rate and lower fertilization rate,implantation rate,clinical pregnancy rate and cumulative live birth rate.It is urgent to establish a clinical prediction model related to infertility of unknown cause to solve the problems of clinical prognosis and individualized medical services,and finally achieve the purpose of increasing the cumulative live birth rate of patients with infertility of unknown cause. OBJECTIVE:To construct and verify the prediction model of high-quality blastocyst formation in patients with unexplained infertility during in vitro fertilization. METHODS:A total of 419 patients with unknown infertility who underwent in vitro fertilization in the Assisted Reproduction Department of Changzhou Maternal and Child Health Care Hospital from March 2017 to June 2022 were retrospectively analyzed,including 317 patients with high-quality blastocysts and 102 patients without high-quality blastocysts.A prediction model was established and used as the model group.The model group was sampled 1 000 times by the Bootstrap method as the validation group.Firstly,the univariate analysis was used to screen the influencing factors of high-quality blastocyst formation of unknown infertility,and the best matching factors were selected by the least absolute shrinkage and selection operator(LASSO)algorithm.Multiple factors were included in the progressive Logistic regression to find out the independent influencing factors and draw a column graph.Finally,the subject working curve,calibration curve,clinical decision curve and clinical impact curve were used to verify the differentiation and accuracy of the prediction model as well as the clinical application efficiency. RESULTS AND CONCLUSION:(1)Univariate analysis of the factors influencing the formation of high-quality blastocyst of unknown infertility were age,insemination method,antimullerian hormone level,basal follicle-stimulating hormone level,basal luteinizing hormone level,human chorionic gonadotropin injection day follicle-stimulating hormone level,human chorionic gonadotropin day estradiol level,progesterone level on human chorionic gonadotropin day,the number of high-quality cleavage embryo(day 3)and the number of blastocyst formation(P<0.05).(2)The best matching factors further screened by LASSO regression were age,insemination method,antimullerian hormone level,basal luteinizing hormone level,human chorionic gonadotropin injection day follicle-stimulating hormone level,human chorionic gonadotropin day estradiol level,the number of high-quality cleavage embryo(day 3)and the number of blastocyst formation(P<0.05).Multifactor stepwise Logistic regression results showed that independent influencing factors on the formation of high-quality blastocysts for unexplained infertility were age,insemination method,antimullerian hormone level,the number of high-quality cleavage embryo(day 3),and the number of blastocyst formation.(3)Receiver operating characteristic curve exhibited that the area under the curve was 0.880(0.834,0.926)in the model group and 0.889(0.859,0.918)in the validation group.It showed that the prediction model had good differentiation.The average absolute error of the calibration curve was 0.036,indicating that the model had good accuracy.The Hosmer-Lemeshow test showed that there was no statistical difference between the prediction probability of blastocyst formation and the actual probability of blastocyst formation(P>0.05).The clinical decision curve and clinical impact curve showed that the model group and the validation group had the maximum clinical net benefit when the threshold probability value was(0.16-0.96)and(0.08-0.93),respectively,and had better clinical application efficacy within the threshold probability range.These findings concluded that age,insemination method,antimullerian hormone,the number of high-quality cleavage embryos(day 3),and the number of blastocyst formation were independent factors influencing the formation of the fine blastocyst in patients with unexplained infertility.The clinical prediction model constructed by these factors has good clinical prediction value and clinical application efficiency and can provide a basis for clinical prognosis and intervention as well as the formulation of individual medical programs.

Effect of Zishen-Yutai pills on refrozen-thawed embryo transfer in patients with diminished ovarian reserve / 中国组织工程研究

BACKGROUND:Currently,hormone replacement therapy is the main treatment in Western medicine for patients with decreased ovarian function,but these patients are not sensitive to exogenous hormones,leading to unsatisfactory therapeutic effect.Zishen-Yutai pills can nourish the blood and calm the fetus,tonify the kidney and spleen,invigorate qi and strengthen the body.Studies have confirmed that Zishen-Yutai pill is effective in reducing follicle-stimulating hormone index and improving traditional Chinese medicine symptoms in patients with diminished ovarian reserve.However,few studies have been conducted to improve the implantation rate of patients by improving endometrial receptivity. OBJECTIVE:To evaluate the effect of Zishen-Yutai pills on the clinical outcome of patients with diminished ovarian reserve undergoing frozen-thawed embryo transfer again. METHODS:A total of 300 patients with diminished ovarian reserve who underwent frozen-thawed embryo transfer to assist pregnancy after the first failure in the Center of Reproductive Medicine,Changzhou Maternal and Child Health Care Hospital from January 2019 to December 2021 were selected as the study subjects.Subjects were randomly treated with a placebo or Zishen-Yutai pills in a ratio of 1:2,with 100 cases in the treatment group and 200 cases in the control group.However,13 patients fell off due to lack of contact,refusal to take medicine or other reasons.Finally,90 cases in the treatment group and 197 cases in the control group were included in the study.Oral medication was administered 7 days before frozen-thawed embryo transfer transplantation at a dose of 5 g/time,3 times/day.To investigate whether taking Zishen-Yutai pills could improve the clinical outcome of patients with diminished ovarian reserve after frozen-thawed embryo transfer again,the primary outcome measures included clinical pregnancy rate,implantation rate,abortion rate,live birth rate,offspring birth weight and birth defects. RESULTS AND CONCLUSION:Compared with the control group,the clinical pregnancy rate(P<0.05)and implantation rate(P=0.009)were significantly increased after the oral administration of Zishen-Yutai pills.Correlation analysis showed that taking the Zishen-Yutai pill was positively correlated with the number of implanted embryos(r=0.200,P=0.001)and clinical pregnancy(r=0.235,P=0.000).There was no correlation between taking Zishen-Yutai pills and indexes of endometrial thickness and blood flow.It is indicated that Zishen-Yutai pills can improve the clinical pregnancy rate and implantation rate of frozen-thawed embryo transfer recurrence in patients with diminished ovarian reserve.

Construction and validation of pregnancy prediction model of artificial insemination by husband based on endometrial structure and uterine spiral artery blood flow parameters / 中国组织工程研究

BACKGROUND:The impact of the endometrium's structure and spiral artery blood flow parameters on the pregnancy rate of artificial insemination by husband remains unclear.This study identified the independent factors and constructed a prediction model with good clinical application efficacy after calibration of other confounding factors. OBJECTIVE:To construct and validate a clinical pregnancy prediction model for artificial insemination by husband based on endometrial structure and uterine spiral artery blood flow parameters. METHODS:A retrospective analysis was conducted on 1 299 patients who underwent artificial insemination by husband treatment at Changzhou Maternal and Child Health Hospital from January 2017 to January 2021.The non-pregnancy group consisted of 1 182 patients,while the pregnancy group included 117 patients.Out of these patients,93 cases were successfully matched between the pregnancy and non-pregnancy groups using a 1∶1 propensity score matching method.Single-factor and multi-factor analyses were used to screen the endometrial structure and uterine spiral artery blood flow parameters to determine their influence on artificial insemination by husband outcomes.The optimal cutoff value was established for each independent influencing factor through receiver operating curve analysis and their risk trend affecting artificial insemination by husband pregnancy outcomes was analyzed using a restricted cubic spline.The clinical efficacy of this combined forecast model was tested by using clinical decision curve and clinical influence curve methods. RESULTS AND CONCLUSION:(1)There was no statistical significance in non-endometrial factors between the pregnancy group and the non-pregnancy group,and the data had a good balance by propensity score matching(P>0.05).(2)Single-factor analysis identified several subendometrial parameters as significant influencing factors of artificial insemination by husband pregnancy outcomes,including vascularization index,flow index,vascular flow index,resistance index,pulsatility index,maximum systolic velocity/end-diastolic velocity,thickness of average junction zone and maximum junction zone from the basal endometrium to the outer myometrium inner layer(P<0.05).(3)Multivariate logistic regression analysis revealed that thickness of average junction zone,pulsatility index,and vascular flow index were independent influencing factors of pregnancy outcomes of artificial insemination by husband,vascular flow index>thickness of average junction zone>pulsatility index.(4)Receiver operating characteristic curve analysis indicated that the area under receiver operating characteristic curve of vascular flow index was 0.704(0.629,0.779),and the optimal cutoff value was 6.26;the area under receiver operating characteristic curve of thickness of average junction zone was 0.660(0.582,0.739),and the optimal cutoff value was 6.38;the area under receiver operating characteristic curve of pulsatility index was 0.642(0.563,0.721),and the optimal cutoff value was 1.18.(5)The restricted cubic spline analysis revealed that artificial insemination by husband pregnancy outcomes were significantly positively affected when the vascular flow index was>6.24 or the thickness of average junction zone was≤6.55 mm,while a negative risk was associated with pulsatility index>1.27.(6)The clinical decision curve and clinical influence curve analyses exhibited that the combined prediction model had the maximum clinical net benefit at the threshold probability value of 0.17-0.93,and the ratio of loss to benefit was consistently less than 1 in the threshold probability range,indicating that the model had good clinical efficacy.(7)It is concluded that after adjusting for other confounding factors outside of the endometrium using propensity score matching and multifactorial logistic regression,the thickness of average junction zone,pulsatility index and vascular flow index were independent factors that influenced pregnancy outcomes of artificial insemination by husband.Through determining their optimal cutoff values and assessing their risk trends,it was confirmed that the combined prediction model had good predictive value and clinical efficacy.

Clinical and genetic analysis of a case with Nicolaides-Baraitser syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology of a girl featuring epilepsy, speech delay and mild mental retardation.@*METHODS@#Peripheral blood samples of the child and her parents were collected. Genomic DNA was extracted and subjected to next generation sequencing. Suspected variant was confirmed by Sanger sequencing.@*RESULTS@#The child was found to carry a de novo heterozygous c.3592G>A (p.V1198M) variant of the SMARCA2 gene, which was predicted to be pathogenic by bioinformatic analysis.@*CONCLUSION@#The child was diagnosed with Nicolaides-Baraitser syndrome due to heterozygous variant of the SMARCA2 gene.

Analysis of pulp revascularization of immature permanent teeth with PRP / 中国基层医药

Objective To discuss the clinical effect of different scaffold in pulp vascular regeneration . Methods 22 young permanent teeth with periapical lesions were randomly divided into observation group and control group,11cases in each group .After the disinfection protocol was completed , the apexes of the control group were irritated to initiate bleeding to produce a blood clot to achieve pulp revascularization ,while the observation group used platelet rich plasma as physical scaffold to operate pulp revasculrization .Results In the control group , the two patients showed painful tooth at 1 and 8 months,the three patients showed healed periapical lesions and narrowed apical foramen at 12 and 18 months ,the six patients showed healed periapical lesions and closure of the apical fora -men at 12 and 18 months.In the observation group ,the three patients showed painful tooth at 1,3 and 8 months,the four patients showed healed periapical lesions and narrowed apical foramen at 12 and 18 months,the four patients showed healed periapical lesions and closure of the apical foramen at 12 and 18 months.There was no statistically significant difference between the observation group and control group by Fisher's exact probabilities (P=1.000). Conclusion Platelet rich plasma is potentially an ideal scaffold for pulp revascularization .

Treatment of mandibular osteoradionecrosis with submental artery island flap and reconstructive Ti-plate / 实用口腔医学杂志

Objective:To investigate the clinical efficacy of reconstruction of the mandibular defect in patients with osteoradionecro-sis using submental artery island flap and reconstructive Ti-plate.Methods:20 cases with mandible osteoradionecrosis underwent par-tial mandibulectomy.The submental artery island flap and reconstructive Ti-plate were used to reconstruct the mandibular defects and adjacent soft tissue defects.The post-operative effects and flap successful rate were evaluated with a follow-up period of 6 to 1 8 months.Results:1 9 flaps were well survived,local necrosis in the remote end was observed in 1 flap,but survived by hyperbaric ox-ygen therapy and iodoform gauze dressing,no plate exposure was found after operation in the follow up period.All patients were satis-factory with the outlook.Conclusion:Submental artery island flap combined with reconstructive Ti-plate is feasible in the treatment of osteoradionecrosis.

Expression and clinical significance of Th17 and Treg cells in peripheral blood of patients with ovarian cancer / 中国免疫学杂志

Objective:To detect the levels of T help cell 17 (Th17)/regulatory T cell (Treg) in the peripheral blood of patients with ovarian cancer and explore their clinical significances.Methods:55 patients with ovarian cancer and 60 normal controls were enrolled.The percentage of Th17/Treg cells were detected by flow cytometry (FCM).The expression of retinoid-related orphan receptor gamma-t ( RORγt)/foxhead winged-helix box protein 3 ( Foxp3) mRNA was measured by real time PCR and the plasma levels of IL-17/TGF-β1 were detected by enzyme linked immunosorbent assay (ELISA).Results: The percentage of CD4+IL-17+/CD4+T cells,the mRNA expression of RORγt and the plasma levels of IL-17 increased significantly in patients with ovarian cancer than in normal controls (P<0.05).Compared with normal controls,the percentage of CD4+Foxp3+Treg/CD4+T cells,the mRNA expression of Foxp3 and the plasma levels of TGF-β1 increased significantly in patients with ovarian cancer ( P<0.05).Conclusion:The imbalance of Th17/Treg cells may exist in the peripheral blood of patients with ovarian cancer.Th17/Treg cells may participate the pathology of ovarian cancer.

Characteristics of GFP-hMPV infection in BALB/c and SCID mice / 中华微生物学和免疫学杂志

Objective To compare the characteristics of hMPV infection in BALB/c and SCID mice.Methods BALB/c and SCID mice were infected intranasally with GFP-rhMPV,and sacrificed on day 3,5,7,9 and 14 post inoculation.Heart,liver,spleen,lungs,kidneys and brain of the animals were used for viral isolation,titration,pulmonary histopathology and detection of GFP-hMPV mRNA expression by RT-PCR and real-time PCR.Results Live viruses were successfully isolated from the lungs of infected mice.Viral titers peaked on the 5th day post inoculation.Viruses remained to be detectable on the 14th day post inoculation in SCID mice,but not in BALB/c mice,whereas genomic RNA of GFP-rhMPV was detectable by PCR targeting F gene in infected BALB/c mice.Live viruses were not able to be isolated from heart,liver,spleen,kidney and brain,neither was genomic RNA of hMPV able to be detected on the 5 th day post inoculation by RT-PCR and real-time PCR.Histopathology of lungs was characterized by interstitial pneumonia on 5 days post inoculation.Lung pathology score of BALB/c mice group was slightly lower than SCID,and the difference was not statistically significant.Conclusion GFP-rhMPV can only replicate in the immunocompetent and immunodeficient mouse lungs,but not in other organs.As compared to that in BALB/c mice,the viral replication appears to be more efficiently and for longer time in SCID mouse lungs probably due to the absence of host cellular and humaral immunity,but this does not necessarily result in more severe pathological lesion.

Molecular cloning and preliminary analysis of TaNOA in common wheat / 生物工程学报

Nitric oxide (NO) is an important signaling molecule with diverse physiological functions in both animal and plant cells. In this work, we isolated the full-length cDNA and genomic DNA sequences of TaNOA-B1 encoding a putative NO associated (NOA) factor in common wheat. Bioinformatic analysis showed that TaNOA-B1 possessed a similar intron/exon structure as its orthologous genes in Arabidopsis and rice. The amino acid sequence deduced from TaNOA-B1 was more than 60% identitical to those of Arabidopsis and rice NOA1 proteins. The primary structure of TaNOA-B1 contained the zinc finger and P-loop GTPase motifs conserved in Arabidopsis and rice NOA1 proteins. There existed at least three NOA gene members in common wheat, which were mapped to homoeologous group six chromosomes 6A, 6B and 6D, respectively. TaNOA-B1 investigated in this work was located on chromosome 6B. The transcripts of TaNOA members were found mainly in leaves. TaNOA-B1-GFP fusion protein may be located in mitochondria. TaNOA transcript level was up-regulated by abscisic acid (ABA) or NaC1 treatments, indicating that TaNOA might be involved in wheat responses to abiotic stresses.

Establishment of plaque assay titration for GFP-labeled recombinant human metapneumovirus / 中华微生物学和免疫学杂志

Objective To establish the plaque assay for the titration of GFP-labeled recombinant human metapneumovims(rhMPV). Methods Vero-E6 cells were selected as host cells for titration. GFP-labeled hMPV was serially diluted and added to each well to infect the cells. The plates were covered with low melting agarose overlay and incubated for different days in incubator. The plates were then observed under fluorescence microscope for plaques with green flourecence, at the same time, the number of plaques was counted by blue plaque-forming. Results Under the low melting agarose overlay, Vero-E6 cells grew well until the CPE caused by hMPV was seen. Clear green flourescence could be observed the first day post infection, much clearer on the third day post infection but showed somewhat fusion between plaques later on.Blue plaques on the fifth day after infection were large and easy to observe. The recombinant GFP-labeled hMPV could replicate up to 1 × 106 PFU in the Vero-E6 cells. Conclusion Plaque assay for titration of recombinant GFP-rhMPV has been sucessfully established. This methodology will offer a solid base for further studies on pathogenesis and vaccine development of this virus.

Molecular and biochemical analysis of two genes encoding dehydroascorbate reductase in common wheat / 生物工程学报

Dehydroascorbate reductase (DHAR) plays an important role in the recycling of ascorbic acid. In this work, we isolated the full length cDNA clones of two different DHAR genes (tentatively named as TaDHAR1 and TaDHAR2, respectively) from common wheat. Semi-quantitative PCR experiments showed that TaDHAR1 and TaDHAR2 were transcribed in many vegetative and reproductive organs examined in this work. Transient expression analysis using wheat protoplasts indicated that the protein products of TaDHAR1 and TaDHAR2 may be located in the cytoplasm. The cDNAs of TaDHAR1 and TaDHAR2 were expressed in the bacterial cells, and resultant histidine tagged recombinant proteins could be efficiently purified using nickel chelate affinity chromatography. In vitro enzyme activity assays revealed that the recombinant TaDHAR1 and TaDHAR2 proteins could all convert dehydroascorbate (DHA) to AsA. The two proteins exhibited higher activity levels at 37 degrees C than at 25 degrees C. Under the two temperature conditions, the optimal pH for TaDHAR1 and TaDHAR2 was both around 7.5. The major difference between TaDHAR1 and TaDHAR2 is the activity under pH 6.0 and 7.0 at 25 degrees C. The results and resources obtained in this study may be useful for further research into the physiological role of TaDHAR genes in AsA metabolism in crop plants under normal or stressed conditions.

联合卒中单元对重症脑卒中影响的综合临床评价 / 中国康复理论与实践

@#目的探讨急性卒中单元联合康复卒中单元对重症脑卒中的影响。方法卒中单元的急性重症脑卒中患者54例为治疗组,神经科常规病房的急性脑卒中患者52例为对照组。比较两组Fugl-Meyger评分、美国卫生研究院脑卒中量表(NIHSS)评分、日常生活活动能力(ADL)评分以及3周感染率、其他并发症发病率、死亡率。结果两组入院时Fugl-Meyger评分、NISHH评分、ADL评分无显著性意义(P>0.05);第3周Fugl-Meyger评分、NISHH评分、ADL评分无显著性差异(P>0.05);第6周出院时或随访两组Fugl-Meyger评分、NISHH评分、ADL评分有非常显著性差异(P<0.01);三周感染率、其他并发症发病率、死亡率有显著性差异(P<0.05)。结论采用急性卒中单元联合康复卒中单元能促进急性重症脑卒中患者运动功能恢复,降低其神经功能缺损程度,提高其日常生活能力。

Influence of the Trial Point on the Quality of Root Canal Obturation / 中国医师杂志

Objective To observe the influence of the trial point on the quality of root canal obturation.Methods 400 teeth were randomly divided into two groups. The experimental group was with the trial point,while the control group without. The quality of obturation was analyzed based on with and without testing the trial point.Results There were 201 teeth with the trial point and 199 teeth without, the rates of flush filled teeth were 86%,75% and 51% respectively in the trial point against,but its rates were 62%,49% and 26% respectively without the test in anterior, premolars and molars. The standard data of two groups were evaluated and compared. The difference of two groups was statistically significant(P

Apoptosis-inducing effect of vp3 gene on human breast carcinoma / 中国普通外科杂志

Objective To construct recombinant plasmid pcDvp3 and observe the apoptosis-inducing effect of vp3 gene on human breast cancer cell line-435. Methods (1) vp3 gene was cloned into the plasmid pcDNA3.1 to form the recombinant plasmid pcDvp3. Then the nucleotides sequencing was processed. (2) 48h after transfection of pcDvp3 and pcDNA3.1 into breast cancer cell lines-435,optical microscopy, electric-microscopy, agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells.(3)Nude mouse model of human breast cancer cells -435 was established to observe the tumor-inhibiting rate and TUNEL was adopted to identify apoptosis. Results (1) Sequence analysis justified the recombination of plasmid pcDvp3. (2) 48h after transfection into breast cancer cells-435, distinct morphological transformation and typical apoptosis bodies were observed, agarose electrophoresis of genomic DNA showed typical ladder-like pattern and flow cytometry analysis showed apoptosis peaks with the percentage of 14.42%. (3) Tumor-inhibiting rates in pcDvp3 groups were 65.52% and 68.23%, much higher than that in pcDNA3.1 group(t=4.06,P