Phylogenetic analysis of chloroplast genome of Tussilago farfara L / 药学学报

italic>Tussilago farfara L. is a perennial herb of Tussilago genus in the Compositae family. Its dried buds and leaves have good biological activities and have a long history of medicinal use in China and Europe. In this paper, we investigated the whole chloroplast genome characteristics, sequence duplication, structural variation and phylogeny of the Tussilago farfara L. After sequencing the Tussilago farfara L. chloroplast genome using Illumination technology, the complete Tussilago farfara L. chloroplast genome was further obtained by assembly and annotation, followed by a series of inverted repeat-large single copy/small single copy region contraction and expansion analysis, genome sequence variation, etc. The sequences of 13 homologous plants downloaded from NCBI were used to construct a neighbor-joining phylogenetic tree. The results showed that the total GC content of the chloroplast genome was 37.4% and the length was 150 300 bp; 125 genes were annotated, including 82 protein-coding genes, 35 tRNAs and 8 rRNAs; 148 (simple sequence repeats, SSR) loci were detected, and the relative synonymous codon usage showed that 31 codons out of 64 codons had a usage of >1. In the phylogenetic analysis, the chloroplast genomes of the seven species of Asteraceae, including the Yulin Tussilago farfara L., were highly conserved, and the sequence variation of the (large single-copy, LSC) and (small single-copy, SSC) regions was higher than that of the (inverted repeat, IR) region. This is in general agreement with the reported phylogeny of Yulin Tussilago farfara L. In this study, we obtained a high quality chloroplast genome and analyzed its genome characteristics, codon preference, SSR characteristics, SC/IR boundary, sequence variation and phylogeny, which can provide a basis for species identification, genetic diversity analysis and resource development of this medicinal plant.

Effect of pH on the chelation between strontium ions and decellularized small intestinal submucosal sponge scaffolds / 北京大学学报(医学版)

OBJECTIVE@#To investigate the preparation of decellularized small intestinal submucosa (dSIS) sponge scaffolds with chelated strontium (Sr) ions at different pH values, and to select the appropriate pH values for synthesizing Sr/dSIS scaffolds using the physicochemical properties and biocompatibility of the scaffolds as evaluation indexes.@*METHODS@#(1) Sr/dSIS scaffolds preparation and grouping: After mixing dSIS solution and strontium chloride solution in equal volumes, adjusting pH of the solution to 3, 5, 7, and 9 respectively, porous scaffolds were prepared by freeze-drying method after full reaction at 37℃, which were named Sr/dSIS-3, -5, -7, and -9 respectively, and the dSIS scaffolds were used as the control group. (2) Physicochemical property evaluation: The bulk morphology of the scaffolds was observed in each group, the microscopic morphology analyzed by scanning electron microscopy, and the porosity and pore size determined, the surface elements analyzed by energy spectroscopy, the structure of functional groups analyzed by infrared spectroscopy, the chelation rate determined by atomic spectrophotometry, the water absorption rate detected by using specific gravity method, and the compression strength evaluated by universal mechanical testing machine.(3) Biocompatibility evaluation: The cytotoxicity and proliferative effect to bone mesenchymal stem cells (BMSCs) of each group were evaluated by Calcein-AM/PI double staining method.@*RESULTS@#Scanning electron microscopy showed that the scaffolds of each group had an interconnected three-dimensional porous structure with no statistical difference in pore size and porosity. Energy spectrum analysis showed that strontium could be detected in Sr/dSIS-5, -7 and -9 groups, and strontium was uniformly distributed in the scaffolds. Functional group analysis further supported the formation of chelates in the Sr/dSIS-5, -7 and -9 groups. Chelation rate analysis showed that the Sr/dSIS-7 group had the highest strontium chelation rate, which was statistically different from the other groups (P < 0.05). The scaffolds in all the groups had good water absorption. The scaffolds in Sr/dSIS-5, -7 and -9 groups showed significantly improved mechanical properties compared with the control group (P < 0.05). The scaffolds in all the groups had good biocompatibility, and the Sr/dSIS-7 group showed the best proliferation of BMSCs.@*CONCLUSION@#When pH was 7, the Sr/dSIS scaffolds showed the highest strontium chelation rate and the best proliferation effect of BMSCs, which was the ideal pH value for the preparation of the Sr/dSIS scaffolds.

Morphological analysis and measurement of spinoglenoid notch of scapula based on 3D reconstruction of CT / 解剖学报

Objective To observe the anatomical structure of spinoglenoid notch of scapula by 3D reconstruction of normal adult scapula by computed tomography (CT), and to provide reference for clinical assessment of suprascapular nerve compression risk, suprascapular nerve compression treatment and shoulder joint operation. Methods Totally 335 cases of normal adult scapula were reconstructed by CT, and classified according to the anatomical shape of spinoglenoid notch; the spinoglenoid notch width (MN), spinoglenoid notch depth (OP), spinoglenoid notch thickness (XY), spinoglenoid notch angle (Z.M0N), distance from 0 point to the inner upper corner of scapula (0A), distance from 0 point to medial lateral edge of scapula (OB), distance from 0 point to inferior angle of scapula (OC) and distance from 0 point to the lowest point of suprascapular notch (OD) were observed and analyzed. Results 1. The morphology of spinoglenoid notch was divided into four types; U type (41. 79%), fin type (42. 99%), L type (8. 36%) and ladder type (6. 86%). U type and fin type were the most common types. Comparison of the four shapes; fin type was the narrowest (11.58 ± 1.74) mm and the deepest (14.58 ± 1.81) mm, the /_ M0N was the smallest (45.62 ± 6.43) ° and the ladder type was the widest (14. 20 ± 2. 67) mm and the shallowest (10. 80 ± 0. 79) mm, the Z.MON was maximum (57. 69 ± 2. 22) ° and the least prone to suprascapular nerve compression. 2. There was no significant difference in MN, OP, XY, zlMON, OA, OB, 0C and OD between left and right sides. 3. The data of MN, OP, XY, OA, OB, OC and OD of men were larger than those of women significantly, but Z. MON was smaller than that of women, indicating that men' s spinoglenoid notch was thicker, wider and deeper, and scapula was wider and longer than that of women. Conclusion The measurement of the morphological and anatomical characteristics of spinoglenoid notch with CT three-dimensional reconstruction is helpful to evaluate the risk of suprascapular nerve compression, the treatment of suprascapular nerve compression, and provide guidance for clinical shoulder surgery.

The prevalence of unhealthy lifestyle among occupational population aged 18‒59 in Shanghai in 2013 / 上海预防医学

Objective:To understand the prevalence of unhealthy lifestyle among Shanghai occupational population aged 18-59 in 2013 and provide suggestion for intervention. Methods:Data retrieved from 2013 Shanghai Non-communicable Diseases and Risk Factors Surveillance were used to investigate the residents who were 18-59 years old and engaged in various occupations in Shanghai. Descriptive analysis and test were conducted by weight and complex sampling. Smoking， drinking， eating behavior and physical activities were compared among different gender，age，area and occupation groups. Results:A total of 10 287 participants were included in this analysis. The current smoking rate of 41.46% in males were significantly higher than 0.64% in females（χ2=2 278.306，P<0.05）. The current smoking prevalence was highest in workers in production and transport （34.49%）. The rate of drinking in the past 30 days， hazardous drinking， and harmful drinking in rural area were 23.80%，6.59% and 11.62%， respectively，which were the highest in all areas. Both the hazardous and harmful drinking prevalence （8.58% and 18.70%， respectively） were the highest in agriculture workers in comparison with other occupations. There is no significant difference in insufficient intake of vegetables and fruits in gender，age and area groups. The proportion of people with excessive intake of red meat was higher in males than that in females （28.11% versus 17.77%，146.176，P<0.05）. Occupational population in urban area had the highest regular physical activities （22.64%）. The clerks had the longest daily sedentary time （7.25 h）. The agricultural workers had the lowest regular exercise rate （11.75%） and the shortest sedentary time （4.07 h）. Conclusion:This study identifies different prevalence of unhealthy lifestyle in different gender， age， area， and occupational groups， which can be helpful for for targeted intervention.

Cystinosis induced by / 中国当代儿科杂志

A boy, aged 1 year and 6 months, was found to have persistent positive urine glucose at the age of 4 months, with polydipsia, polyuria, and growth retardation. Laboratory examinations suggested that the boy had low specific weight urine, anemia, hypokalemia, hyponatremia, hypomagnesemia, metabolic acidosis, glycosuria, acidaminuria, increased fractional excretion of potassium, and decreased tubular reabsorption of phosphate. X-ray examinations of the head, thorax, and right hand showed changes of renal rickets. The slit-lamp examination showed a large number of cystine crystals in the cornea. The genetic testing showed a suspected pathogenic homozygous mutation of the

First batch of national demonstration areas for comprehensive prevention and control of chronic diseases in Shanghai:their healthy environment construction and effectiveness evaluation / 上海预防医学

Objective To make assessment on construction effectiveness of the first batch of national chronic disease comprehensive control and prevention demonstration areas (NCD demonstration areas) in Shanghai, summarizing NCD experience and its characteristics, so as to provide reference for other regions. Methods Data were collected and analysed concerning construction of health support environment and regional characteristics of the six districts-Xuhui, Jing′an, Minhang, Jiading, Songjiang and Qingpu.Descriptive data from Shanghai Non-communicable Diseases and Risk Factors Surveillance in 2013 and 2017 was also analyzed for evaluation. Results It was noted that health support environment had been in continuous construction.The total number of health units increased by 1 502 as compared with pre-construction.The number of new type health units increased by 489 such as health theme parks, health trails and health huts.All six districts established self-help health test points covering all streets and towns.Residents′ living and behavior styles were improved, as there was a decrease in current smoking rate, oil intake and salt intake in the six NCD demonstration areas.In Xuhui, Minhang and Jiading three districts, the current smoking rate among residents over 18 years old remarkably declined.In five of the six districts, the average daily oil intake per family member was significantly reduced and so was the average daily salt intake per family member in the six districts.In addition, the six demonstration areas in Shanghai explored several models of NCD prevention and control in line with regional characteristics, which produced positive results. Conclusion The first batch of demonstration areas of Shanghai have achieved effective results in aspects of environmental support and health effects.The modes of NCD prevention and control in different districts present their regional characteristics, which provide significant reference for other regions of Shanghai in this regard.

Consumption of sugar-sweetened carbonated beverage and juice/fruit-flavored drink among residents aged 15 years and above in 2013 in Shanghai / 上海预防医学

Objective To determine the consumption of sugar-sweetened carbonated beverage and juice/fruit-flavored drink among residents aged 15 years and above in 2013 in Shanghai. Methods Data was extracted from the 2013 Shanghai Non-communicable Disease and Risk Factors Surveillance, in which multi-stage cluster sampling was performed.A total of 25 657 subjects aged ≥15 years across the city were selected for analysis. Results In the study, 34.42% (95%CI:32.33%-36.51%) and 37.85% (95%CI:35.37%-40.32%) of the subjects consumed sugar-sweetened carbonated beverage and juice/fruit-flavored drink in 2013 in Shanghai.The proportions declined with age (P for trend < 0.001), while increased with education level (P for trend < 0.001).They were 65.28% and 69.82% among students, which were much higher than other occupations.Men consumed more sugar-sweetened carbonated beverage than women (37.31% vs 31.37%), whereas less juice/fruit-flavored drink (36.14% vs 39.65%).Among the subjects consumed sugar-sweetened carbonated beverage (n=6 254) or juice/fruit-flavored drink (n=6 701), 52.20% (95%CI:49.41%-54.98%) and 54.04% (95%CI:51.09%-56.98%) consumed 1-3 times a week.The daily average amount of sugar-sweetened carbonated beverage and juice/fruit-flavored drink were 98.64 mL (95%CI:88.92-108.37) and 88.85 mL (95%CI:73.73-103.97), respectively, which were higher among the young (< 45 years old), male and students. Conclusion In 2013 in Shanghai, the consumption of sugar-sweetened carbonated beverage and juice/fruit-flavored drink is highly prevalent among residents aged 15 years and above, especially among the young, male, well-educated and students.Intervention should be tailored to reduce the consumption among at-risk population.

Research progress on pharmacological properties and monoterpenoid indole alkaloids biosynthetic pathway in plants of genus Rauvolfia / 中草药

Rauvolfia serpentine is a traditional Indian medicine and now the genus Rauvolfia becomes a commonly used south medicine in China. Rauvolfia is mainly used to treat hypertension, mental illness, cardiovascular disorders, cancer, and other diseases. The paper overviewed the main species and distribution, pharmacological properties, and the monoterpenoid indole alkaloids biosynthetic pathway of Rauvolfia in China. The paper aims to provide reference for research and clinical application in plants of the genus Rauvolfia.

Therapeutic effect of lentivirus-mediated miR-21 on chemotherapy-induced premature ovarian failure in rats / 解放军医学杂志

Objective To explore the therapeutic potential of miR-21 in rat model of chemotherapy-induced premature ovarian failure (POF).Methods Lentivirus-mediated miR-21 (LV-miR-21) was constructed successfully in vitro with molecular biology methods.Rats were divided into 4 groups named control group,model group,blank vector group and miR-21 group.Rat models of chem0therapy-induced POF were established in the latter 3 groups by intraperitoneal injection ofcytoxan (CTX).Bilateral ovaries of rats in miR-21 group were injected with LV-miR-21,of rats in blank vector group were injected with lentivirus vector,and rats in model group received no treatment.At 1,15,30,45 and 60 days after the last injection,blood sample was collected,the rats were then sacrificed and the ovaries were removed.The estrous cycle was observed by vaginal smears.The E2 level was detected by chemiluminescent immunoassay,and the follicle stimulating hormone (FSH) level was detected by homologous double antibody radiation immunoassay.Ovary weights were measured,and the follicle count was conducted through observing paraffin section under microscope.The apoptosis of ovarian granulosa cells was analyzed by TUNEL assay.Results During 15-30 days,30-45 days and 45-60 days after the last injection,regular estrous cycle was recovered respectively in 8,5 and 3 rats in miR-21 group.At the 15th,30th,45th and 60th day after the last injection,the E2 level was higher in miR-21 group than in model group and blank vector group,but the FSH level showed the opposite trend (P=0.000).At the 45th and 60th day after the last injection,the follicle numbers at all stages increased markedly in miR-21 group than in model group and blank vector group (P=0.000).At the 30th,45th and 60th day after the last injection,the ovary weights were higher in miR-21 group than in model group and blank vector group.At the 15th,30th,45th and 60th day after the last injection,the apoptosis rate of ovarian granulosa cell were significantly lower in miR-21 group than in model group and blank vector group (P=0.000).Conclusion Up-regulation of miR-21 expression may partly recover the ovarian structure and function damaged by CTX.

A study on immunologic response at different periods with different hepatitis B vaccine in infants / 预防医学

Objective To know the immunologic response at different periods with different hepatitis B vaccine(HepB)in infants.Methods From July 1 to December 31 in 2009,the local infants born from hepatitis B virus(HBV)surface antigen (HBsAg)negative mother in Ninghai were selected and received a 0.5 ml of 5 -μg HepB made by recombinant deoxyribonucleic acid (DNA)techniques in saccharomyces cerevisiae yeast (HepB -SCY)and those in Yuyao were selected and received a 0.5 ml of 10 -μg HepB made by recombinant DNA techniques in hansenula polymorpha yeast (HepB -HPY)with a 0 -1 -6 month schedule.The specimens of blood were collected at 7 months,12 months and 18 months after vaccination for testing antibody.Results At 7 months,12 months and 18 months after vaccination,the geometric mean concentration (GMC)of Anti -HBs in HepB -SCY group were 714.79 ±3.50 mIU /mL,259.89 ± 3.87 mIU /mL and 78.38 ±4.04 mIU /mL,respectively.While in HepB -HPY group were 2 420.76 ±3.51 mIU /mL, 703.55 ±3.46 mIU /mL and 171.32 ±3.76 mIU /mL,respectively.There was statistical significant difference of GMC between HepB -SCY group and HepB -HPY group(P <0.01).The percent of antibody <100 mIU /mL in HepB -SCY group were 6.39%,23.33% and 55.42%,and in HepB -HPY group,were 2.17%,5.17% and 31.95%,respectively. The difference between HepB -SCY group and HepB -HPY group had significantly statistical significance (P <0.01 ). Conclusion The immunologic response at different periods of HepB -HPY was better than that of HepB -SCY.

Correlation of fluid resuscitation volume with intra-abdominal hypertension and renal injury in patients with severe acute pancreatitis / 第二军医大学学报

Objective To investigate the correlation of fluid resuscitation volume with intra-abdominal hypertension (IAH) and renal injury in patients with severe acute pancreatitis (SAP). Methods Totally 60 patients with SAP from our ICU were recruited in the present study. The intra-abdominal pressure (IAP) was measured by method AbViser. The patients were divided into the following 3 groups according to the different of IAP: the normal group, IAH group and abdominal compartment syndrome (ACS) group. The general information, mortality rate, fluid resuscitation volumes and renal injury were analyzed and compared in the 3 groups. Results (1) The incidence rates of IAH and ACS in our study were 31.67%(19/60) and 11.67% (7/60), respectively. The mortality rates of normal group, IAH group and ACS group were 5.88%(2/34), 21.05%(4/19) and 57.14%(4/7), respectively. The mortality rate of ACS group was significantly higher than that of normal group (P=0.007). (2) The fluid balance volumes of normal group, IAH group and ACS group were (4 350±892), (5 512±1 246) and (5 974±1 765) mL, respectively, with significant differences found among the 3 groups (P<0.001); the total cumulative fluid volumes of the 3 groups were (8 530±2 384), (9 403±2 064) and (13 172±2 409) mL, respectively, also with significant differences found among the 3 groups (P<0.001). ACS group had the highest fluid balance and total cumulative fluid volumes in the first 3 days. (3) There were significant differences in the levels of blood creatinine and urea nitrogen among the 3 groups (P<0.001). The levels of blood creatinine and urea nitrogen of ACS group was significantly higher than those of normal group and IAH group (P<0.05). Conclusion The cumulative fluid of the first 3 days is associated with IAH and even ACS in pateints with SAP, and IAH can further lead to renal injury in patients with SAP.

Effect of modified sijunzi decoction on the bone metabolism of adriamycin induced nephropathy rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Modified Sijunzi Decoction (MSD) on the bone metabolism of prednisone intervened adriamycin-induced nephropathy rats.</p><p><b>METHODS</b>The adriamycin-induced nephropathy rat model was prepared. Totally 50 SD rats were randomly divide into five groups, i.e., the model group, the hormone group, the Chinese medicine (CM) group, the CM + hormone group, and the normal control group. The 24-h urine samples were collected on the 7th, 21st, and 35th day after modeling. The 24-h urine protein was measured by biuret colorimetry. Serum levels of osteoprotegerin (OPG), receptor activator of nuclear factor-kappaB ligand (RANKL), osteocalcin (BGP), and tartrate-resistant acid phosphatase (TRACP) were determined by ELISA. Expressions of OPG and RANKL in the tibia tissue were detected using real-time quantitative PCR and Western blot.</p><p><b>RESULTS</b>(1) Compared with the normal control group, the 24-h urine protein increased in each group on the 7th, 21st, and 35th day (P < 0.05, P < 0.01). Compared with the model group, the 24-h urinary protein decreased in the hormone group and the CM + hormone group (P < 0.05, P < 0.01). The decrement was more obvious along with the treatment time went by (P < 0.05, P < 0.01). There was statistical difference in the reduction of urine protein on the 35th day between the CM group and the model group (P < 0.05). (2) Compared with the 21st-day of the same group, the serum levels of TRACP and RANKL increased (P < 0.05, P < 0.01). Compared with the model group, the serum levels of the TRACP and RANKL increased (P < 0.05, P < 0.01), OPG and BGP decreased (P < 0.05, P < 0.01) in the hormone group. Compared with the CM group at the same period, serum OPG level decreased and the RANKL level increased in the hormone group and the CM + hormone group (P < 0.05, P < 0.01). Besides, the serum level of TRACP increased and BGP decreased (P < 0.05, P < 0.01). Compared with the hormone group at the same period, OPG and BGP increased (P < 0.05, P < 0.01), RANKL decreased (P < 0.01) in the CM + hormone group. On the 35th day TRACP decreased (P < 0.01). (3) Compared with the normal group, mRNA expressions of OPG and RANKL on the 21st day increased (P < 0.05, P < 0.01), mRNA expressions of OPG and RANKL on the 35th day decreased in the model group (P < 0.01). Compared with the CM group at the same period, OPG mRNA expression decreased (P < 0.01) and RANKL mRNA expression increased in the hormone group (P < 0.05). OPG mRNA expression decreased in the CM +hormone group (P < 0.05). (4) Compared with the hormone group on the 21st day, the OPG level decreased and the RANKL protein increased (both P < 0.05). RANKL decreased in the CM + hormone group (P < 0.05). Compared with the model group at the same period, OPG decreased and RANKL increased in the hormone group (P < 0.01). Compared with the CM group at the same period, OPG decreased (P < 0.01), RANKL increased (P < 0.01) in the hormone group and the CM + hormone group. Compared with the hormone group at the same period, OPG increased and RANKL decreased in the CM + hormone group (both P < 0.01).</p><p><b>CONCLUSIONS</b>Prednisone could induce osteoporosis through the OPG/RANKL/RANK pathway. MSZ could slow down the formation of prednisone-induced osteoporosis through promoting osteoblast differentiation, and inhibiting osteoclastogenesis.</p>

Isolation, culture and identification of human odontoclasts / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To isolate, culture and identify odontoclasts in vitro and to establish a method of culturing human odontoclasts.</p><p><b>METHODS</b>Healthy and retentive deciduous teeth were extracted, and then placed in α-minimum essential medium containing 0.1% collagenase and 0.2% dispase for 1 h.Odontoclasts were obtained and incubated from the absorbing root surfaces of deciduous teeth.Isolated cells were viewed by inverted phase contrast microscope firstly. Then, the isolated odontoclasts were morphologically observed by hematoxylin and eosin staining (HE) and tartrate-resistant acid phosphatase (TRAP) staining. The prepared teeth slices were cocultured with the isolated odontoclasts and scanning electronic microscope(SEM) was used to demonstrate the presence of resorption lacunae.</p><p><b>RESULTS</b>The isolated odontoclasts appeared as multinucleated giant cell with many vacuolus in cytoplasm. TRAP staining demonstrated that the cytoplasm of the odontoclasts was full of claret-red positive particles.Resorption lacunae on teeth slices which cocultured with odontoclasts were seen under SEM.</p><p><b>CONCLUSIONS</b>Enzyme digestion is an effective method to isolate odontoclasts from absorbing root surface of deciduous teeth.</p>

Renal expression of macrophage migration inhibitory factor in children with Henoch-Sch-nlein purpura nephritis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To examine the expression of macrophage migration inhibitory factor (MIF) in renal tissues obtained from children with Henoch-Sch-nlein purpura nephritis (HSPN).</p><p><b>METHODS</b>The renal tissue samples were obtained from 11 children with different pathological grades of HSPN and 8 children with thin glomerular basement membrane disease (controls). The MIF expression was measured by immunohistochemistry. The correlation between MIF expression and 24 hrs urinary protein excretions was evaluated using a linear correlation analysis.</p><p><b>RESULTS</b>MIF expression was seldom found in renal tissues obtained from controls. However, a significantly increased MIF expression was found and was concordant with the increased severity of renal pathology in renal tissues obtained from children with HSPN. The MIF expression in renal tissues of grade III-IV of renal pathology was significantly higher than that in grade I-II in children with HSPN (p<0.01). In children with HSPN, there was an increased MIF expression in renal tissues with crescent formation and inflammatory cell infiltration. Renal MIF expression was significantly positively correlated with 24 hrs urinary protein excretions in children with HSPN (p<0.01).</p><p><b>CONCLUSIONS</b>MIF may play an important role in renal injury of HSPN. Up-regulation of MIF expression may reflect the degree of renal lesions in HSPN.</p>

Relationship between leukotrienes and clinical, pathological features of Henoch-Schoenlein purpura nephritis in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the role of leukotrienes (LTs) in the progress of children's Henoch-Schoenlein purpura (HSP) nephritis (HSPN), and to provide an experimental basis for clinical application of leukotrienes antagonists in treatment of HSPN.</p><p><b>METHODS</b>The serum and urine samples were collected from 34 patients with HSPN 18 of them received renal biopsy, 27 cases with HSP and 16 healthy children as control. The level of LTB4 in the serum and urine was tested by enzyme-linked immunosorbent assay (ELISA) and LTE4 in urine in each group by enzyme immunoassay (EIA). The extent of expression of LTC4 synthase was detected by indirect immune fluorescence (IIF) in the renal tissue of 18 HSPN cases who received renal needle biopsy. Meanwhile, 3 cases with thin basement membrane nephropathy (TBMN) and 4 cases with simple hematuria were enrolled as controls. The results of pathological examination of the 4 cases with simple hematuria was normal by light microscope or electron microscope. At the same time, total urine protein in 24 hours was determined in 24 HSPN patients.</p><p><b>RESULT</b>(1) The level of serum and urinary LTB4 in the children with HSPN was (1164.33 +/- 300.28) ng/L and (841.19 +/- 115.23) ng/L, respectively. The level of serum and urinary LTB4 in those with HSP was (559.60 +/- 180.23) ng/L and (574.42 +/- 101.17) ng/L, respectively. The level of serum and urinary LTB4 in the control group was (211.95 +/- 67.72) ng/L and (227.33 +/- 76.12) ng/L, respectively. There was significant difference in the LTB4 level between HSPN group and HSP group (P < 0.01) while there was statistically significant difference in the LTB4 level between HSPN group and control group (P < 0.01). The urinary LTE4 was (1252.31 +/- 251.62) ng/L, (805.93 +/- 185.52) ng/L and (149.51 +/- 33.66) ng/L for HSPN group, HSP group and control group, respectively, and the differences were significant (P < 0.01). (2) The increase of serum and urinary LTB4 and urinary LTE4 was closely relative to the severity of histopathological changes. (3) Serum and urinary LTB4, urinary LTE4 increased in parallel to the enhancement of urine protein in HSPN patients (P < 0.01 or P < 0.05). (4) Markedly significant difference of LTC4 synthase by IIF existed between HSPN groups and control group.</p><p><b>CONCLUSION</b>LTs can promote the progress of children's HSPN. There is a close relationship between LTs expression in renal tissues, the pathological severity of HSPN and proteinuria.</p>

Construction of selectable marker-removable plant expression vectors / 生物工程学报

The commonly used plant constitutive expression vector pBI121 was modified by insertion of two directly orientated lox sites each at one end of the selectable marker gene NPTII and by replacing the GUS gene with a sequence composed of multiple cloning sites (MCS). The resulting plant expression vector pBI121-lox-MCS is widely usable to accommodate various target genes through the MCS, and more importantly to allow the NPTII gene removed from transformed plants upon the action of the Cre recombinase. In addition, the CaMV 35S promoter located upstream of the MCS can be substituted with any other promoters to form plant vectors with expression features specified by the introduced promoters. Provided in this paper is an example that an enhanced phloem-specific promoter of the pumpkin PP2 gene (named dENP) was used to construct an NPTII-removable phloem-specific expression vector pBdENP-lox-MCS. Moreover, to facilitate screening of selectable marker-removed gene and the composite sequence is flanked by lox sites. Thus the selectable marker-free plants can be visually identified by loss of GFP fluorescence. The above newly created plant expression vectors can be used to develop selectable marker-removable transgenic plants for a variety of purposes.

Construction,purification and substrate specificity identification of recombinant human platelet-activating factor acetylhydrolase isoformⅠ / 上海第二医科大学学报

Objective To construct and purify the recombinant protein of platelet-activating factor acetylhydrolase(PAF-AH) isoformⅠ,and study the enzyme activity by different substrates. Methods The ? subunit of PAF-AH isoformⅠwas cloned and expressed in E.coli.Exogenously expressed recombinant protein was purified to SDS-PAGE homogeneity,and its activity was identified by arylesterase detection.Phenylacetate,l-O-hexadecyl-2-deoxy-2-thioacetyl-sn-glycero-3-phosphocholine(2-Thio PAF) and 1-myristoyl-2-(4-nitrophenylsuccinyl) phosphatidylcholine(the latter two were commercial plasma PAF-AH substrates) were used for the substrate identification.The plasma type PAF-AH was served as positive control. Results Recombinant protein of ? subunit of PAF-AH isoformⅠwas successfully constructed and expressed in E.coli after purification.Compared with positive control,the recombinant protein could hydrolyze phenylacetate and 2-Thio PAF,but could not hydrolyze 1-myristoyl-2-(4-nitrophenylsuccinyl) phosphatidylcholine.Conclusion Recombinant protein of ? subunit of PAF-AH isoformⅠcan be successfully constructed.There are differences in the substrate specification to the two commercial PAF substrates for PAF-AH isoformⅠand plasma type PAF-AH,which provides a quick method to differentiate PAF-AH isoformⅠfrom plasma type PAF-AH.

Using green fluorescent protein as a reporter to monitor elimination of selectable marker genes from transgenic plants / 生物工程学报

In genetic modification of plants, once the transformants are obtained, selection markers are no longer required in mature plants. At present, the Cre/lox site-specific recombination system is most widely used to eliminate the selectable marker genes from the transgenic plants. In this study, attempt was made to favour the selection of marker-free plants in the re-transformation method. Green fluorescent protein (GFP) can be directly visualized in living cells, tissues or organisms under UV illumination. This advantage of GFP is exploited in the development of a practical approach in which GFP is used as a visual marker to monitor the removal of the selectable marker gene from transgenic plants. For that purpose, the pGNG binary vector was constructed, in which the GFP gene (gfp) was linked to the expression cassette Nos P-nptII-NosT and the two units were cloned between two directly-orientated lox sites. The CaMV 35S promoter was placed before the first lox site and used to drive GFP expression. The beta-glucuronidase gene (gus) of Escherichia coli was cloned behind the second lox site without a promoter, thus would not be expressed in this position. Tobacco plants were first transformed with pGNG and selected on kanamycin (Kan)-containing media. Regenerated transgenic shoots were readily singled out by GFP fluorescence. The GFP-expressing plants were then re-transformed with pCambia1300-Cre containing hygromycin phosphotransferase gene (hpt) as a selectable marker gene. The Cre-mediated recombination resulted in the elimination of lox-flanked genes, herein gfp and nptII, from the plant genome and brought the GUS gene next to the 35S promoter. Our data demonstrated that transgenic plants free of nptII were easily selected by monitoring the loss of green fluorescence, and at the same time, GUS (here as a target protein) was expressed in the nptII-free plants. Finally, hpt and cre were removed from the progenies of the nptII-free plants by gene segregation.

Methylprednisolone and cyclophosphamide pulse therapy of severe systemic lupus erythematosus in children / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the effect of methylprednisolone (MP) and cyclophosphamide (CPA) intermittent intravenous pulse therapy and the clinical prognosis in children with severe juvenile onset systemic lupus erythematosus (JOSLE).</p><p><b>METHODS</b>Thirty patients with JOSLE, diagnosed by clinical, laboratory or renal histological examinations, were enrolled in this study. Of the 30 patients, 27 were females and 3 were males, the mean age was (12 +/- 3) years, and 20 of the 22 patients who had undergone initial therapy had LN, and the clinical courses before being involved in the study were 3 to 12 months in nine patients. Twenty-three of the 30 patients had clinical manifestations of renal damages, of whom 4 patients were proven by initial renal biopsy to have WHO type IV, 2 had type II,1 had type V and 1 had type III, and 7 patients had one or more manifestations of central nervous system, including chorea, seizures, cerebrovascular accident (CVA) and organic brain syndrome (OBS), simultaneously, 9 patients had nervous system symptoms without the clinical manifestations of renal damages, 3 patients had lupus crisis, 7 patients did not have any manifestations of renal or neurological damages. According to the protocol of the therapy, the patients were divided into 3 groups: group A (n = 18) patients were treated with MP plus CPA intermittent intravenous pulse for children with lupus nephritis, and with or without neuropsychiatric lupus erythematosus (NPLE), group B (n = 7) with pulsed doses of MP, followed by prednisone and tripterygium wilfordii hook f(T(whf)) for patients without renal or central nerves system damage, and group C (n = 5) with prednisone alone for patients with LN determined by clinical and laboratory features. The effects of those regimes and the clinical prognosis were observed.</p><p><b>RESULTS</b>On short-term follow-up, the SLEDAI-2K (by weight of the renal damage) showed significant difference between group A and group B, but there was no significant difference at the 9th months of the therapy. The long-term follow-up lasted in average for (37.2 +/- 24.8) months. Nineteen patients were followed up for more than 18 months. At the end of follow-up, the mean age was 14 to 19 years. There was no difference on the effect of both group A and group B, and no frequent infections were seen, ANAs were negative and SLEDAI-2K = 0-point in two patients of each group 12 months after discontinuation of the therapy. Four patients in group C died within 18 months.</p><p><b>CONCLUSION</b>The immunosuppressive regimen MP + CPA in patients with severe JOSLE and MP + prednisone + T(whf) in patients without major organs damage were superior to the regimen of prednisone alone.</p>