Development and application of two novel functional molecular markers of BADH2 in rice

BACKGROUND: Fragrance is one of the most important quality traits in rice, and the phenotype is attributed to the loss-of-function betaine aldehyde dehydrogenase (BADH2) gene. At least 12 allelic variations of BADH2 have been identified, and some of these have been applied to rice fragrance breeding using traditional molecular markers and Sanger sequencing techniques. However, these traditional methods have several limitations, such as being very expensive, imprecise, inefficient, and having security issues. Thus, a new molecular marker technology must be developed to improve rice fragrance breeding. RESULTS: In this study, more than 95% of the cultivated fragrant rice varieties belonged to a 7-bp deletion in exon 2 (badh2-E2) or an 8-bp deletion and 3-bp variation in exon 7 (badh2-E7). Both allelic variations resulted in the loss of function of the badh2 gene. We developed two novel SNP molecular markers, SNP_badh2-E2 and SNP_badh2- E7, related to the alleles. Their genotype and phenotype were highly cosegregated in the natural variation of rice accessions, with 160 of the 164 fragrant rice varieties detected with the two markers. These markers cosegregated with the fragrance phenotype in the F2 population. CONCLUSIONS: Two functional SNP molecular markers of badh2-E2 and badh2-E7 allelic variations were developed. These functional SNP molecular markers can be used for genotype and genetic improvement of rice fragrance through marker-assisted selection and will significantly improve the efficiency of fragrant rice breeding and promote commercial molecular breeding of rice in the future.

Prenatal diagnosis and neonatal phenotype of a de novo microdeletion of 17p11.2p12 associated with Smith–Magenis syndrome and external genital defects

Smith–Magenis syndrome (SMS, OMIM: 182290) is a multiple congenital anomalies and intellectual disability syndrome due to a 3.45 Mb microdeletion involving 17p11.2 and is estimated to occur about one in 25,000 births. Up to now, the ultrasound findings of the foetus with SMS and their external genital defects in patients are rarely reported. This case indicates that foetus with SMS may present polyhydramnios and ventriculomegaly in the second trimester. The newborn male patient had an abnormal phenotype in which he has micropenis and his anus is close to the perineal body. The identification of this case may further expand the phenotypic spectrum of this genetic disorder.

Study on Protective Mechanism of Panax Notoginseng Saponins on Rats with Renal Ischemia Reperfusion Injury Based on Klotho / 中国中医药信息杂志

Objective To investigate the effects of Panax Notoginseng saponins (PNS) on protein expression of Klotho in rats with renal ischemia reperfusion injury; To discuss its protective mechanism for model rats. Methods Experimental rats were randomly divided into sham-operation group, model group, positive medicine group, PNS high-, medium- and low-dosage groups. Each administration group was given relevant medicine for gavage, once a day. Renal ischemia reperfusion injury model was established. Rats were sacrificed by taking blood from abdominal aorta after 4 hours of modeling. Serum levels of blood urea nitrogen (BUN), creatinine (SCr), malondialdehyde (MDA) content in kidney tissue, superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) activity were measured. HE staining was used to observe the morphological changes of renal tissue. The protein expressions of Klotho and NF-κB p65 were measured by immunohistochemical method. Results Compared with the sham-operation group, the levels of BUN and SCr in the model group increased significantly (P<0.05); protein expression of Klotho in renal tissue decreased and the protein expression of NF-κB p65 increased (P<0.05). Compared with the model group, the expression of Klotho increased but protein expression of NF-κB p65 decreased in each administration group (P<0.05); Compared with the positive medicine group, the expression of Klotho in PNS high-dosage group increased but protein expression of NF-κB p65 decreased (P<0.05). The protein expression of NF-κB p65 was negatively related to protein expression of Klotho (r=-0.895, P<0.05). Conclusion PNS can inhibit oxidative stress and anti-inflammatory effects through upregulating protein expression of Klotho, and reduce the protein expression of NF-κB p65, and thus exerts renal protective effects.

Osthole enhances doxorubicin-induced apoptosis in p53-wildtype prostate cancer cells by down-regulating SIRT1 expression / 中国病理生理杂志

AIM:To investigate the effect and mechanism of osthole on increasing the cytotoxicity of doxorubi -cin(DOX)to prostate cancer cells.METHODS:MTT assay was performed to evaluate the viability of LNCaP cells trea-ted with osthole and DOX.The protein expression of silent information regulator 1(SIRT1),p53,acetylated p53 and Pu-ma,as well as release of cytochrome C and activation of caspase-9 and caspase-3 in the LNCaP cells treated with osthole and DOX were determined by Western blot.The apoptosis of the LNCaP cells treated with osthole and DOX was analyzed by flow cytometry.RESULTS:Osthole significantly increased the cytotoxicity of DOX against p 53-wildtype prostate cancer cell line LNCaP.Osthole significantly inhibited the expression of SIRT 1 in the LNCaP cells.Transfection with SIRT1 plas-mid decreased the cytotoxicity of osthole and DOX co-treatment against LNCaP cells.Combination with osthole and DOX significantly induced the over-expression and acetylation of p53.Transfection with p53 siRNA significantly decreased the synergistic effect of osthole on cytotoxicity of DOX-treated LNCaP cells.Combination with osthole and DOX significantly in-duced the release of cytochrome C into the cytoplasm from mitochondria,followed by activation of caspase-9 and its down-stream molecule caspase-3,thus leading to cell apoptosis in the LNCaP cells.CONCLUSION:Osthole promotes the p53-dependent apoptosis in DOX-treated prostate cancer LNCaP cells by down-regulating the expression of SIRT1.

Pulmonary toxicity of single-wall carbon nanotubes and its oxidative stress mechanism / 第二军医大学学报

Objective To systematically study the pulmonary toxicity of single-wall carbon nanotubes (SWCNTs) and to explore the related cytotoxicity mechanism, so as to provide a theoretical basis for the safe productionandapplication of SWCNTs. Methods A549 cells were cultured in the media containing 0, 25, 50, 100, 150, and 200 μg/mL SWCNTs for 24 h, and then the cell viability and degree of cell membrane damage were assessed by CCK-8 and lactate dehydrogenase (LDH) release assay kit, respectively; the ultrastructural alteration of A549 cells was detected by transmission electron microscope (TEM). The oxidative stress response was evaluated by assessing reactive oxygen species (ROS), glutathione (GSH) and superoxide dismutase (SOD). The rats were exposed to SWCNTs by intratracheal inhalation, and then the animals were sacrificed 3 days later and the pathological sections of lung tissue were examined. Results SWCNTs showed considerable toxicity to A549 cells, decreasing cell viability, causing severe damage of cell membrane and ultrastructure, increasing the intracellular ROS level, and decreasing GSH content and SOD activity. It was found that oxidative stress is the main mechanism of SWCNTs toxicity on A549 cells. In vivo toxicity results showed that SWCNTs accumulated in the lung tissue # causing alveolar wall edema. Conclusion In vitro and in vivo toxicity results have found that SWCNTs possess a significant pulmonary toxicity # with its main toxicity mechanismbeing oxidative stress.

Association of MDR1 gene polymorphisms with the risk of hepatocellular carcinoma in the Chinese Han population

The multidrug resistance 1 gene (MDR1) is an important candidate gene for influencing susceptibility to hepatocellular carcinoma (HCC). The objective of the present study was to evaluate the association of MDR1 polymorphisms with the risk of HCC in the Chinese Han population. A total of 353 HCC patients and 335 healthy subjects were enrolled in the study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), created restriction site-PCR (CRS-PCR) and DNA sequencing methods were used to identify MDR1 gene polymorphisms. Two allelic variants (c.335T>C and c.3073A>C) were detected. The CC genotype of the c.335T>C polymorphism was associated with an increased risk of developing HCC compared to the TT genotype (OR = 2.161, 95%CI = 1.350-3.459, χ2 = 10.55, P = 0.0011). The risk of HCC was significantly higher for the CC genotype in the c.3073A>C polymorphism compared to the AA genotype in the studied populations (CC vs AA: OR = 2.575, 95%CI = 1.646-4.028, χ2 = 17.64, P < 0.0001). The C allele of the c.335T>C and c.3073A>C variants may contribute to the risk of HCC (C vs T of c.335T>C: OR = 1.512, 95%CI = 1.208-1.893, χ2 = 13.07, P = 0.0003, and C vs A of c.3073A>C: OR = 1.646, 95%CI = 1.322-2.049, χ2 = 20.03, P < 0.0001). The c.335T>C and c.3073A>C polymorphisms of the MDR1 gene were associated with the risk of occurrence of HCC in the Chinese Han population. Further investigations are needed to confirm these results in larger different populations.

Review of the clinical features of skin cancer: a study of 1, 048 cases from a hospital in West China

Skin cancer [SC] is a group of malignancies which include primary and metastatic tumors which involve the skin and its appendages. Up to the present, only a few studies on the clinical features and the trend of S have been reported but the status in West China is still undetermined. The S cases were from a major hospital in West China. A total of 1048 cases from 1981 to 2006 were included in our study. The clinical features of S including age, gender, lesion location and pathological diagnosis were analyzed. In order to illustrate the trend of S incidence, the patients from 1981-1993 and 1994- 2006 were assigned to group A and B respectively. The percentage of S in all malignancies [Ms], including all kinds of internal carcinomas and skin cancers, and the percentage of S in inpatients and outpatients [IOPs] between group A and B were separately compared to illustrate the trend in S incidence in this area. [1] Of the 1048 S s included, 308 [29.4%] were squamous cell carcinoma [S C], 293 [28.0%] basal cell carcinoma [B] and 168 [16.0%] cutaneous malignant melanoma [MM] .Ratio of male to female was 1.5:1.0.Median age was 54.0 +/- 23.0 years.40.8%of the S s occurred on the head, 35.0%on the trunk and 24.2%on the extremities. Median age of MM [53.0 +/- 22.5] was less than those of B [58.0 +/- 18.3 years] and S [57.0 +/- 20.0 years] .83.6%of the B s, 49.8%of the S s and 13.5% of the CMMs occurred on the head. [2] Of the 168 MMs, 106 [63.1%] occurred on the acral, 23 [13.7%] on the head, 24 [14.3%] on the trunk and 15 [8.9%] on the limbs. Of the 106 acral melanoma [AM], 41 [38.7%] occurred on the plantar skin, 19 [17.9%] on the heel, 15 [14.2%] on the subungual skin of thumbs, 13 [12.3%] on the subungual skin of big toes and 18 [17.0%] on other acra. [3] The percentages of S in IOPs [S s/IOPs] in Group A and B were 0.0038% [325/8, 457, 672], 0.0066% [723/11, 037, 720], an increase of by 74%.The percentages of S in all Ms [S /Ms] were 2.1% [325/15, 363] and 3.1% [723/23, 364], an increase of 48%.During the same period, the percentages of Ms in IOPs [Ms/IOPs] were 0.18% [15, 363/8, 457, 672] and 0.21% [23, 364/11, 037, 720], increased only by 17%. In our study, S C, B and MM were major S types. The head and trunk are the main sites for S occurring. AM is the most common MM. In past 26 years, the percentages of S in all malignancies and in inpatients and outpatients have increased in this hospital. The finding in our study provides a clue for understanding of the trend of S in West China