Construction and validation of sheep VASA gene knock-in vector based on CRISPR/Cas9 system / 生物工程学报

This study aimed to explore the expression changes of VASA gene in sheep testis development and to construct VASA gene knock-in vector to prepare for the study on the differentiation of sheep germ cells in vitro. The testicular tissues of 3-month-old (3M) and 9-month-old (9M) sheep which represent immature and mature stages, respectively, were collected. The differential expression of VASA gene was analyzed by quantitative real-time PCR (qPCR) and Western blotting, and the location of VASA gene was detected by immunohistochemistry. The sgRNA targeting the VASA gene was designed and homologous recombination vectors were constructed by PCR. Subsequently, plasmids were transferred into sheep ear fibroblasts. The VASA gene was activated in combination with CRISPR/dCas9 technology to further verify the efficiency of the vector. The results showed that the expression level of VASA gene increased significantly with the development of sheep testis (P < 0.01), and was mainly located in spermatocytes and round spermatids. The knock-in vector of VASA gene was constructed by CRISPR/Cas9 system, and the Cas9-gRNA vector and pEGFP-PGK puro-VASA vector were transfected into ear fibroblasts. After CRISPR/dCas9 system was activated, ear fibroblasts successfully expressed VASA gene. The results suggest that VASA gene plays a potential function in sheep testicular development and spermatogenesis, and the VASA gene knock-in vector can be constructed in vitro through the CRISPR/Cas9 system. Our results provided effective research tools for further research of germ cell development and differentiation.

Improvement of asthenopia caused by abnormal visual functions via visual training in adult patients / 中华实验眼科杂志

Objective:To analyze the improving effect of visual therapy on the symptoms of asthenopia caused by abnormal visual function in adults.Methods:A serial case study was conducted.Ninety-three adult patients (186 eyes) with visual dysfunction caused by abnormal visual function who underwent training in the visual training room of the Optometry Center of Tianjin Eye Hospital from October 2018 to October 2019 were enrolled, among which there were 48 males and 45 females.The average age of patients was (30.43±6.39) years old.Binocular visual function examination included vision examination by phoropter, simultaneous vision and stereopsis by Worth 4 Dots test, distance and near heterophoria by Von-Graefe method test, fusion range by rotating prism method, the accommodation reaction by fusion cross cylindrical lens (FCC), the accommodation amplitude by minus technique, the accommodation flexibility and the vergence flexibility by flipper, and a personalized training program was formed on the basis of the above examination results.The training process was divided into initial examination, first review and second review, and each stage containing 5 times of training was followed by a review.The results of the initial examination, first review and second review were compared.The study protocol was approved by an Ethics Committee of Tianjin Eye Hospital (No.KY201906). Written informed consent was obtained from each patient prior to any examination.Results:The average near heterophoria was -8.0 (-15.3, -3.0) △ at the initial examination, and it was reduced to -5.0 (-9.0, 0.0) △ at the first review, showing significant difference (Z=-3.586, P<0.01). The mean accommodation amplitude of left and right eyes were 4.00 (3.25, 5.25)D and 4.00 (3.00, 5.00)D respectively before visual training, which were increased to 5.50 (4.25, 7.00)D and 5.00 (3.75, 7.00)D at the first review, showing significant differences (Z=-4.284, -3.995; both at P<0.01). The broken point and the recovery point of the long-distance positive fusion at the initial inspection were 7.5 (5.0, 15.8) △ and 0.0 (0.0, 4.0) △ respectively, which were increased to 11.0 (6.0, 22.0) △ and 4.0 (0.0, 7.0) △ respectively at the first review, showing significant differences (Z=-3.192, -3.748; both at P<0.01). The broken point and the recovery point of the near positive fusion at the initial inspection were 18.0 (8.0, 28.0) △ and 6.0 (0.0, 12.0) △, respectively, which were increased to 26.0 (21.5, 35.0) △ and 11.5 (6.0, 16.0) △ respectively at the first review, showing significant differences (Z=-4.695, -3.377; both at P<0.01). The monocular and binocular accommodation flexibility were increased from 2-3 cycles/minute at the initial examination to 10-12 cycles/minute at the first review, showing significant differences (all at P<0.01). Logistic regression analysis showed that age, distance heterophoria and near heterophoria were not related to the difference in accommodation improvement of the right and left eyes.At the initial examination, the average CISS score of 22 patients was (25.13±9.64) points, which was dropped to (19.18±7.22) points at the first review, showing significant difference ( t=6.79, P<0.01). The 67.60% (48/71) of the patients who did not answer the questionnaire had obvious improvement in their main complaints and physical signs, and 29.58% (21/71) of them had improvement but still needed more training, and 2.82% (2/71) had no improvement in visual fatigue symptoms. Conclusions:Systemic visual therapy can improve the visual function and alleviate symptoms majority of the adult patients with abnormal visual function, suggesting that adult visual function is still of strong plasticity.

Long-term effects of GnRHa on central precocious puberty or early and fast puberty children / 中华内分泌代谢杂志

Objective:To investigate the long-term effects of GnRHa treatment on final height gain, gonadal function, and body mass index(BMI) in children with central precocious puberty(CPP) or early and fast puberty(EFP), and to explore the influencing factors of height gain and early predictors.Methods:Fifty patients with CPP and 44 patients with EFP who were treated with GnRHa for more than 2 years were enrolled(80 females and 14 males). Body height, bone age, BMI, gonads hormone, uterus and ovarian volumes(female), testicular volume(male), and other parameters before and after treatment were measured.Results:(1)For girls: GnRHa plus GH treatment gained more final height compared with GnRHa treatment [(10.69±5.73) cm vs (7.42±5.76) cm, P<0.05]. Height lost >5cm at the initial treatment benefited much more for the final height compared with height lost<5cm [(10.65±3.32) cm vs (6.51±3.40) cm, P<0.01]. The proportion of overweight/obesity decreased when reaching the final height compared with the initial treatment and stopping the treatment. Serum LH level, uterine and ovarian volume were significantly decreased after stopping treatment compared with before treatment, and increased half a year to 1 year after stopping treatment.100% of girls had menarche and 95% reached the regular cycle 3 years after stopping treatment.(2)For boys: GnRHa plus GH treatment and GnRHa treatment gained height by(8.78±5.2) and(7.99±4.82) cm, respectively. Serum LH level and testicular volume were significantly decreased after stopping treatment as compared with those before treatment, and increased for half a year to 1 year after stopping treatment. Conclusion:GnRHa treatment can significantly improve the final height for girls with CPP and EFP. The patients with more height lost could gain more height, which can be used as a predictor of height gain.

Application of digital droplet PCR in detection of activating hotspot mutations in GNAS gene in children with McCune-Albright syndrome / 中华内分泌代谢杂志

Objective:To detect the known hotspot mutations of GNAS in children with McCune-Albrigtht syndrome(MAS) by droplet digital PCR, and to explore its application value in the diagnosis of MAS.Methods:A total of 122 children with MAS were enrolled in the pediatric department of Ruijin Hospital Affiliated to Medical College of Shanghai Jiaotong University. For the known mutation hotspot of GNAS gene (R201H/C), dd-PCR, real-time fluorescent pyrophosphatic activation polymerase reaction (PAP) and second-generation sequencing were used to detect the presence of gene mutation and to analyse the relevance with the clinical features.Results:GNAS gene mutation was detected in 89 out of 122 children with MAS and 57 cases were found to have mutations. The positive rates of ddPCR, PAP, and second generation sequencing were 77.42%, 29.03%, and 56.25%, respectively. The GNAS gene mutation was detected in all classical triad patients. Among them, the positive rates of ddPCR in peripheral blood of typical and atypical children were 100% and 73.1% respectively, which were significantly higher than those of the other two methods. The detection rate of GNAS mutation in precocious puberty with bone lesions was higher than that in precocious puberty with skin lesions, suggesting that fibrous dysplasia with precocious puberty is an important basis for clinical diagnosis of MAS in children.Conclusion:Precocious puberty is the most common endocrine manifestation of MAS in children. Bone fibrous dysplasia with precocious puberty is an important factor in clinical diagnosis. ddPCR has high sensitivity, which can be helpful for molecular diagnosis of MAS.

Long-term effects of GnRHa on central precocious puberty or early and fast puberty children / 中华内分泌代谢杂志

Objective@#To investigate the long-term effects of GnRHa treatment on final height gain, gonadal function, and body mass index(BMI) in children with central precocious puberty(CPP) or early and fast puberty(EFP), and to explore the influencing factors of height gain and early predictors.@*Methods@#Fifty patients with CPP and 44 patients with EFP who were treated with GnRHa for more than 2 years were enrolled(80 females and 14 males). Body height, bone age, BMI, gonads hormone, uterus and ovarian volumes(female), testicular volume(male), and other parameters before and after treatment were measured.@*Results@#(1)For girls: GnRHa plus GH treatment gained more final height compared with GnRHa treatment [(10.69±5.73) cm vs (7.42±5.76) cm, P<0.05]. Height lost >5cm at the initial treatment benefited much more for the final height compared with height lost<5cm [(10.65±3.32) cm vs (6.51±3.40) cm, P<0.01]. The proportion of overweight/obesity decreased when reaching the final height compared with the initial treatment and stopping the treatment. Serum LH level, uterine and ovarian volume were significantly decreased after stopping treatment compared with before treatment, and increased half a year to 1 year after stopping treatment.100% of girls had menarche and 95% reached the regular cycle 3 years after stopping treatment.(2)For boys: GnRHa plus GH treatment and GnRHa treatment gained height by(8.78±5.2) and(7.99±4.82) cm, respectively. Serum LH level and testicular volume were significantly decreased after stopping treatment as compared with those before treatment, and increased for half a year to 1 year after stopping treatment.@*Conclusion@#GnRHa treatment can significantly improve the final height for girls with CPP and EFP. The patients with more height lost could gain more height, which can be used as a predictor of height gain.

Efficacy and safety of letrozole in treatment of McCune-Albright syndrome girls with peripheral precocious puberty / 浙江大学学报·医学版

OBJECTIVE@#To evaluate the efficacy and safety of the third-generation aromatase inhibitor letrozole in the treatment of McCune-Albright syndrome (MAS) girls with peripheral precocious puberty.@*METHODS@#Twenty-one MAS girls with peripheral precocious puberty treated in Pediatrics Department of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from March 2012 to June 2017 were enrolled in the study. Patients presented with repeated vaginal bleeding, premature breast enlargement, café-au-lait spots or dysplasia of bone fibers, and low levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH); and the congenital adrenal hyperplasia, estrogen-producing tumors, and exogenous estrogen intake were excluded. Letrozole were administrated at a dose of 0.5-2 mg·m ·d for 6 to 12 months. The patients were observed for changes in breast staging, vaginal bleeding, sex hormone levels, liver function and bone age changes, and changes in uterine and ovarian volume.@*RESULTS@#After treatment, bone age/chronological age (BA/CA)was decreased from 1.23±0.30 to 1.11±0.18 ( < 0.01); the predicted adult height (PAH) increased from (156.2±5.9)cm to (158.4±2.1)cm after treatment ( < 0.05); the vaginal bleeding was reduced and the estradiol level decreased, while the teststosterone level and the uterus showed no significant increase, and no adverse reactions such as ovarian torsion and abnormal liver function were observed.@*CONCLUSIONS@#Precocious puberty is one of the most common endocrine manifestations in MAS. Our findings suggest that letrozole may be an effective and safe therapy to precocious puberty in girls with McCune-Albright Syndrome.

Application and evaluation of moral education materials of scientific research cases in the teaching of Experimental Traditional Chinese Medicine / 中华医学教育探索杂志

Objective:To evaluate the teaching effect of moral education materials implied in scientific research cases in the teaching of "Experimental Traditional Chinese Medicine (TCM)".Methods:The moral education materials implied in scientific research cases of "Experimental TCM" were extracted and skillfully integrated into the teaching of professional knowledge. A questionnaire survey was conducted with questions as "whether it is good to demonstrate the potential humanistic spirit by case teaching, whether this helps improve your interest in science and whether this teaching form affects your study, life and work attitude in the future" "Ten specific items from scientists' moving deeds that touch students and their recognitions" to assess the teaching effect.Results:95.8 percent of students affirmed this teaching form and thought it helped improve their interest in scientific exploration. 87.5 percent of students considered the humanistic spirit would affect their study, life and work in the future. 77.1-89.6 percent of students held a positive attitude to the ten items derived from the scientists' moving stories. Among these items, the percentages of the two items, "the spirit of being able to endure loneliness, work hard to make contributions selflessly without seeking for rewards or reputations" and "having respect forpeople and their scientific research achievements with courage to challenge the authorities of scientific research" are the lowest and highest, respectively.Conclusion:It is good to apply the moral education materials implied in the scientific research cases in the teaching of "Experimental TCM", which basically achieves the teaching goal, but there is still room for improvement.

Practice and reflections on the teaching reform of Experimental Traditional Chinese Medicine guided by scientific research cases / 中华医学教育探索杂志

Experimental Traditional Chinese Medicine is an emerging discipline that plays an important role in cultivating innovative talents of traditional Chinese medicine (TCM). In recent years, with the rapid development of TCM and the new requirements of positioning, and also combined with the college students' cognitive characteristics, we have revised and republished the "Experimental TCM" (Third Edition) textbook, which focuses on introducing knowledge by adopting relevant scientific research cases. This test book was used in the teaching of undergraduates of batch 2013 in the eight-year program in Shanghai University of Traditional Chinese Medicine. After-class questionnaires showed that this teaching mode, guided by scientific research case, is not only helpful for students to develop their quality and ability of adopting modern experimental methods initially in the study and development of TCM, but also able to spread the great achievements of TCM researches. The teaching mode is also conducive to enhancing students' sense of responsibility for the modernization of TCM. Therefore, it is suggested that the course of Experimental TCM should be promoted in the colleges and universities of TCM.

Changing trend regarding the burden on cerebrovascular diseases between 1990 and 2016 in China / 中华流行病学杂志

Objectives@#To analyze the status quo and trends on the burden of cerebrovascular diseases between 1990 and 2016 in China.@*Methods@#Morbidity mortality, years of life lost (YLL), years of lived with disability (YLD) and disability-adjusted life year (DALY) related to cerebrovascular diseases between 1990 and 2016, were collated and analyzed, according to the results of the Global Burden of Diseases Study 2016 (GBD 2016). Numbers on incidence and morbidity were used to assess the incidence of diseases, while the numbers of death and mortality were used to assess the death of diseases. Years of life lost due to premature death (YLL), years lost due to disability (YLD) and disability-adjusted life year (DALY) were used to assess the burden of diseases. Changing trend on the burden of cerebrovascular disease from 1990 to 2016 was also analyzed.@*Results@#In 2016 and 1990, the numbers of new cases/morbidity and the number of deaths/mortality on cerebrovascular diseases in the country showed an upward trend. Rates regarding YLL and DALY on cerebrovascular diseases remained stable from 1990 to 2016, however, the YLD rate showed a slow upward trend. The changing rate of DALY was mainly influenced by YLL. Both DALY and YLL crude rates in males showed a slow upward trend, with the highest DALY rate appearing in the ≥70 age group. Disease burden on males was heavier than that of the females and in the 50-60 age group, which taking the largest proportion. As for the composition in DALY, YLL appearing much larger than YLD and slowly increasing. Analysis on the subtypes of diseases, proportions of YLL and DALY in hemorrhagic stroke were greater than that in ischemic stroke while the proportion of YLD in ischemic stroke was in the opposite.@*Conclusions@#The burden of disease on cerebrovascular diseases remained heavy and the differences appeared in age, gender and subtypes of diseases. Our findings called for the adoption of measures including screening, intervention and rehabilitation to be taken on target populations, in order to reduce the burden on both individuals and the society.

Analysis of the etiology and clinical characteristics of short stature / 临床儿科杂志

Objective To explore the etiology and clinical characteristics of short stature. Method Clinical data of 2075 children with short stature treated from May 1995 to July 2017 were retrospectively analyzed. The etiology and morbidity of pathological short stature and normal variant short stature were analyzed. The clinical characteristics of growth hormone deficiency (GHD) , idiopathic short stature (ISS) , constitutional delay in growth (CDG) and familial short stature (FSS) were analyzed. The etiological differences between severe short stature [height standard deviation score (SDS) ≤-3] and general short stature (height SDS＞-3) were analyzed. Results Among 2075 children diagnosed with short stature, 1719 (82.84％) were pathological short stature, among which GHD (38.60％) and ISS (22.02％) were more common. Normal variant short stature was found in 356 children (17.16％) , with FSS and CDG accounting for 10.70％ and 6.46％ respectively. There were statistically significant differences in the sex ratio, age at initial diagnosis, height SDS, body mass index (BMI) , bone age and bone age delay among children with four common childhood short stature (GHD, ISS, CDG and FSS) (all P＜0.01) . Boys were more than girls in four kinds of childhood short stature. The height SDS was the lowest in GHD group and the highest in CDG group; BMI was highest in GHD group, but lower in CDG and ISS group. Bone age delay was highest in GHD group and lowest in CDG group. In severe short stature group, the rates of complete GHD, multiple pituitary hormone deficiency, small for gestational age infant, Turner syndrome, hypothyroidism and Russell-Silver syndrome were higher than those in general short stature group, but the rates of partial GHD, ISS, FSS and CDG was lower than those in general short stature group. Conclusion The etiology of short stature is complex. Analysis of the etiology and clinical features is helpful for clinical diagnosis and treatment.

A study of congenital lipoid adrenal hyperplasia in two pedigrees / 中华内分泌代谢杂志

The clinical features and laboratory data of two patients with congenital lipoid adrenal hyperplasia (CLAH)were collected. The genomic DNA was extracted from the peripheral blood white cells in the two patients and their family members and the STAR gene was screened for mutations by PCR and Sanger sequencing. Patient 1 was a girl aged 2 years and 8 months,and she visited us because of continual cyanosis for more than two years. Physical examination showed no obvious pigmentation or clitoral hypertrophy,and Tanner stage was B1P1. Clinical examination revealed serum ACTH 1 284.1 pg/ml and 17α-hydoxyprogesterone(17-OHP)0.54 ng/ml, with Karyotype 46, XX. Genetic analysis showed compound heterozygous mutations of c.201_202delCT and c.229C>T in the STAR gene. Her father carried heterozygous c.201_202delCT mutation, and her mother showed heterozygous c.229C>T mutation. Patient 2 was a girl aged 22 years and referred to us because of dark skin for more than 21 years. Physical examination revealed generalized hyperpigmentation,with Tanner stage B5P2. Hormone examination showed ACTH>2 000 pg/ml and serum cortisol 0.77μg/dl. Karyotype analysis revealed 46,XX. Genetic analysis found compound heterozygous mutations of c.64+1G>C and c.707_708delinsCTT in the STAR gene,which descended from her father and mother respectively. Of note,c.64+1G>C is a novel splicing mutation of STAR gene.

Four cases of PTPN11 associated RASopathies:Analysis of the correlation between gene mutation and clinical phenotype / 中华内分泌代谢杂志

PTPN11 is the most common mutation gene of RAS disease, which is located in the upstream of RAS/MAPK pathway and participates in signal transduction. Because the molecular mechanism of RAS's disease involves the same pathway, it may present a certain commonality in clinic, but the different genotypes with PTPN11 mutation may also express different phenotypes. Therefore, it is not easy to identify and diagnose this disease early in clinic. The present article aims to analyze the correlation between the clinical phenotype and genotype of 4 patients with RAS disease.

Genetic diagnosis of erythrocyte pyruvate kinase deficiency and discovery of PKLR gene novel mutations / 中华实用儿科临床杂志

Objective To investigate the clinical symptoms of pyruvate kinase deficiency (PKD) and the new mutation type of PKLR gene in 3 cases of PKD,and to explore the method for PKD gene diagnosis.Methods Sequencing of blood system-related genes in 3 children was performed by target sequence capture and high-throughput sequencing technology,and the protein function of mutant gene was forecasted,after detecting the pathogenicity of the patients,these genotypes were confirmed by Sanger sequencing.Results In the 3 children,5 types of PKLR gene mutations were found:double heterozygous mutations c.1529G ＞ A(p.R510Q) and c.1031T ＞ G(p.I344S),homozygous mutation c.847G ＞ T (p.V283F),double heterozygous mutations c.979delC (p.L327fs)and c.604_617del (p.V202fs).PKLR gene c.1529G ＞ A(p.R510Q) mutation had been reported previously,and the other four mutations were new.c.1031T ＞ G (p.I344S) and c.847G ＞ T (p.V283F) was possibly pathogenic mutation,which meant that the probability of mutation of this gene was more than 90％ and c.979delC (p.L327fs) and c.604 _617del (p.V202fs) variation was a pathogenic variation.These 5 mutations had a greater effect on protein function,and all ofthem were pathogenic mutations.Conclusion Since PKD patients are difficult to be diagnosed clinically,PKLR gene variation can be detected by target sequence capture and high throughput sequencing technology,and the pathogenicity of the new mutant is evaluated.

A multi-center clinical study of early predictors and follow-up parameters for girls with rapidly progressive central precocious puberty / 中华内分泌代谢杂志

Objective To study the early diagnostic predictors and key follow-up parameters for girls with rapidly progressive central precocious puberty (RP-CPP). Methods A total of 260 girls with CPP participated in a prospective, nonrandomized, multi-center, nested case control study. After follow-up six months without any therapy, 114 girls were divided into RP-CPP (n=70) and slowly progressive CPP (SP-CPP) (n=44) groups. Results The basal serum LH and insulin-like growth factor Ⅰstandard deviation score (IGF-ⅠSDS) were the important risk factors of RP-CPP (OR 4.04, 1.578), especially the former. The receiver operating characteristic (ROC) curve revealed that the areas under the ROC curve of basal LH and IGF-ⅠSDS were 0.83 and 0.807, respectively. The levels of basal LH and IGF-ⅠSDS were at 0.52 mIU/ml and 0.35 respectively for the accuracy diagnosis of RP-CPP with the maximum Youden indexs. After follow-up for six months, the change levels of height, breast stages, bone age/chronological age ratio, serum LH, uterine and ovarian volume in RP-CPP group were significantly higher than those in SP-CPP group (all P<0.05). Conclusions The level of basal serum LH and IGF-ⅠSDS may be used as the risk predictors for early diagnosis for girls with RP-CPP. The change levels of basal LH, progress rates of gonad and sex character, height, and impaired growth potential seem to be the key follow-up parameters for CPP progress.

Analysis of clinical features and related genes variation in 41 girls with McCune-Albright syndrome / 中华内分泌代谢杂志

Objective To investigate the clinical characteristics and molecular pathological mechanism of McCune-Albright syndrome ( MAS) in order to provide a guidance for the precision medicine of MAS. Method The clinical data and genetic findings of 41 patients with MAS were analyzed retrospectively. Results (1) MAS girls had the phenotype of peripheral precocious puberty with premature sexual development and high estradiol, low LH and FSH, and the increased volume of uterus and ovary. ( 2 ) In 41 MAS cases, there were 17 cases with GNAS1 gene mutation, and the total positive rate was 41. 5%, of which the classic triad was 66. 7%, two signs 56. 3%, and 12. 5% in only one classic sign. GNAS1 gene mutation was found in 78. 6% of patients with polyostotic fibrous dysplasia of bone, while only 55. 0% in patients with cafe au lait skin spots. Children with precocious puberty and fibrous dysplasia of bone is an important basis for clinical diagnosis of MAS, but cafe au lait skin spots does not seem to be the specifical manifestation of MAS. Conclusion Clinically MAS was lack of typical clinical manifestations, and the most important clinical weight factor for the diagnosis of MAS was peripheral precocious puberty with fibrous dysplasia of bone. GNAS1 gene screening may be helpful to improve the clinical accurate diagnosis of MAS.

Analysis of clinical and physical factors for acute bone marrow suppression induced by concurrent chemoradiotherapy in rectal cancer / 中国肿瘤临床

Objective:To analyze the clinical and physical tactors associated with acute bone marrow suppression in concurrent chemo-radiotherapy for rectal cancer and to provide a reference standard for the best clinical treatment plan. Methods:Retrospective analy-sis was performed on 62 patients with rectal cancer who received concurrent radiotherapy and chemotherapy in our department. The pelvis was contoured for each patient in the radiotherapy treatment planning system and divided into three subsites: lumbosacral spine, ilium, and lower pelvis. Prognostic clinical and physical factors were analyzed by univariate and multivariate analyses. Evaluated prognostic clinical factors included sex, age, clinical stage, original hemoglobin levels, and chemotherapy, operation, and radiation modes;physical factors included V5, V10, V15, V20, V25, V30, V35, V40, V45, V50, Dmax, and Dmean of lumbosacral spine, ilium, low-er pelvis, and pelvis. Results:The percentage of patients who developed acute bone marrow suppression (≥2 grade) was 61.3%(38/62).Univariate analysis of related factors revealed statistically significant differences were sex, chemotherapy, lumbosacral spine V45, il-ium V20, and ilium V30. Multivariate logistic regression analysis indicated that chemotherapy and ilium V30 are the risk factors for acute bone marrow suppression. The receiver operating curve showed that the threshold of ilium V30 was 44%. Conclusion:Acute bone marrow suppression is influenced by more than one factor;local control rate of the tumor and acute bone marrow suppression are tradeoffs in rectal cancer treatment. An appropriate chemotherapy method should be selected, and ilium V30 must be maintained below 44%to prevent bone marrow suppression in rectal cancer patients.

Identification of pelvic dose-volumetric parameters that predict acute bone marrow suppression in concurrent chemoradiotherapy for cervical cancer / 中华放射医学与防护杂志

Objective To identify the dose-volumetric parameters associated with acute bone marrow suppression in concurrent chemoradiotherapy for cervical cancer,and provide the reference standard for the radiotherapy planning.Methods In total,155 patients concurrently receiving chemoradiotherapy for cervical cancer were enrolled in this study.The pelvis was contoured for each patient in radiotherapy treatment planning system and divided into three subsites:lumbosacral spine,ilium,and lower pelvis.The pelvic dose volume parameters were analyzed using univariate analysis (Chi-Square and t test),and multivariate analysis (Logistic regression model).Dose volume parameters include V5,V10,V15,V20,V25,V30,V35,V40,V45,V50 and the average dose (D).Results The percentage of patients that developed acute bone marrow suppression (≥ grade 2) was 87.7％ (136/155).The univariate analysis revealed that pelvic V5,V10,V15,V,0,ilium V15,lower pelvis V15 of patients with acute bone marrow suppression (≥ grade 2) were significantly higher than those of ＜ grade 2 patients (t =-2.277,-2.142,-3.475,-2.018,-2.963,-2.741,P ＜0.05).Multiple regression analysis indicated that pelvic V15 was associated with acute bone marrow suppression (OR =1.728.P ＜ 0.05).The threshold of pelvic V15 was 88％ as determined by receiver operating curve (ROC).Conclusions The results show that Pelvic V15 is associated with acute bone marrow suppression in concurrent chemoradiotherapy for cervical cancer and is thus an independent risk factor.To better predict and control acute bone marrow suppression,pelvic V15 should be carefully controlled below 88％ in treatment planning to reduce the incidence of acute bone marrow suppression.

The Role of Metallothioneins (MTs) in the Occurrence and Development of Hepatocellular Carcinoma and Mechanism Behind the Prevention and Treatment of Traditional Chinese Medicine / 世界科学技术-中医药现代化

It is a brief review of our research on the role of metallothioneins (MTs) in the occurrence and development of hepatocellular carcinoma (Hcc) in more than a decade,and the mechanism of prevention and treatment for Hcc in the field of TCM.It was found that MTs were required for the malignant proliferation of Hcc cells and their expressions significantly increased under different levels of stress.MTs may exert their effects on promoting proliferation via increasing ribosomal protein gene or other gene expressions.The over-expressions of MTs directly altered the balance of ions in the cell nucleus,and interacted on the genes promoting malignant proliferation.Hepatotoxicity and hepatic damage caused by DEN were reduced and relieved to different extents by the representative formula or constitute,which under the common therapeutic principle,by which MTs expressions were down-regulated as well.In conclusion,both effects above are important in the liver protection,inhibition of hyperplasia and carcinogenesis.

Effect of tumor size on the expression of adrenal cholesterol homeostasis molecules in H22 hepatoma-bearing mice / 中国实验动物学报

Objective To study the effect of tumor size on the expression of adrenal cholesterol homeostasis mole -cules in H22 hepatoma-bearing mice.Methods Two hundred and twenty mice were injected with H 22 hepatoma cells to their right armpit.On the 11th day after injection, the mice were sorted according to the tumor size .18 mice with large tumor (large tumor group) and 18 mice with small tumor (small tumor group) were sacrificed, and the tumors were weighed.A control group consisting of 18 normal Kunming mice was also included in this study .The plasma TC, TG and HDL-C were detected using total cholesterol , triglycerides or HDL-C assay kits , respectively .The mRNA expressions of Srb1, Ldlr, Npc1, Npc2, Stard3, Hmgcr, Lipe, Acat1, Abca1, Abcg1, Srebp-1c, Lxrα, Lxrβ, Rxrα, Apoa1 and Apoe were tested by real-time quantitative RT-PCR ( qRT-PCR ) , and Gapdh and β-actin were used for normalization .SRB1 and ApoA1 proteins were analyzed by Western blot .Results The tumor weight was significantly higher in the large tumor group than that in the small tumor group (P<0.05).Compared with the control group , the plasma HDL-C was significant-ly decreased in the two hepatoma groups (P<0.05).The expression levels of Srb1, Ldlr, Apoa1 mRNA and SRB1 protein were significantly increased in the large tumor group (P<0.05 for all).The ApoA1 protein level was significantly higher in the large tumor group than that in the small tumor group (P<0.05).The expressions of Acat1, Lipe, Abca1 and Abcg1 mRNA were significantly lower in the large tumor group than those in the small tumor group (P<0.05 for all).However, the expressions of Srebp-1c, Lxrαand RxrαmRNA were not significantly changed , then, Srebp-1c, Lxrβand RxrαmRNA expressions were significantly up-regulated in the small tumor group (P<0.05).The expressions of Hmgcr and Apoe mR-NA were not significantly different in the two groups .Conclusions In hepatoma-bearing mice , due to the adaptation to tumor-induced chronic stress response , the adrenal cortical cells can effectively utilize intracellular cholesterol to synthetize cortical hormones .

Exploration and evaluation of the literature studying and reading route in PBL teaching of the courses of TCM integrated experiments / 中华医学教育探索杂志

Objective On the issues of students' lack of representative collection of literature in problem-based learning (PBL) courses of TCM integrated experiments, to try to explore the litera-ture studying and reading route. Method Totally 41 full-time seven-year medical postgraduate of Shanghai University of TCM were randomly divided into two PBL classes, that are class A (n=20) an class B (n=21). Three PBL units were carried out in class A and class B by parallel; the oriented literatures were provided by tutors alternately, and after PBL courses, the appraisal were performed by students and tutors independently, using SPSS 19.0 for data analysis, t test to compare between groups. Results First, the students' assessment indicated that the 75% (15/20) students of Class A and 80.9%(17/21) students of Class B of class gave a good evaluation of the teaching method. Two classes had more than 80% of the students who thought the literature teachers provided was representative and instructive and over 60.0% students of both classes thought that the teaching measures could improve the classroom efficiency and after-school learning efficiency. The tutors' assessment revealed that there were no significantly differences between the quantity of these references in PPT (P>0.05) made by students in A class and B class in PBL unit 2 and unit 3; but an increasing trend did exist after the leading literatures were provided and the discussions among students were focusing and rele-vant and the teaching goal was achieved . Conclusion The route of providing a small amount of oriented literatures by tutor is positive in PBL courses of TCM integrated experiments, while the design of teaching experiment, the specific way of providing oriented literatures, the quantity and quality of literatures, and even the internal connection between them, all needs further optimization.