Dynamic DNA circuits have shown promising potential for amplified biosensing and bioengineering applications at the molecular level. Here, an enzyme-free, single-step and rapid signal amplification DNA circuit was developed by integrating target-directed entropy-driven catalysis (EDC) and hybridization chain reaction (HCR) for analysis of nucleic acids and small molecules. The target catalyzes the self-assembly of the EDC premade substrate complex and fuel strands to release the hidden amplicon trigger (T), which was encoded with trigger sequences for the downstream HCR circuit. The released T could motivate the successive cross-opening of HCR hairpins yielding long DNA nanowires and generated tremendously amplified fluorescence signals. Notably, this EDC-HCR circuit was driven by entropy without the requirement of any enzymes, thus greatly reducing the cost. The design of the hidden amplicon trigger (T) avoided the production of waste by-products and improved the reaction rate. Furthermore, as a modular circuit, we also demonstrated that our EDC-HCR cascade sensing system could be used as a versatile sensing platform for the highly sensitive and selective detection of other analysts, e.g. ATP in serum samples, through simply programming the reorganization sequences of the initiator. Therefore, the flexible and versatile EDC-HCR platform holds great potential in the fields of clinical diagnosis and biochemical analysis.
A dinuclear complex [(ppy)Ir(tpy)PtCl]2+ (Ir–Pt) can exhibit strong antitumor activity towards cisplatin-resistant cancer cells and induce cell necrosis via mtDNA damage and mitochondrial dysfunction.
Post-traumatic stress disorder (PTSD) is closely related to the exposure to traumatic events and results in the structural and functional changes of hippocampus. Human basic helix-loop-helix family member e40 (BHLHE40) was reported to be implicated with neuron maturity and neuronal differentiation. The present study aimed to reveal the role of BHLHE40 on single-prolonged stress (SPS) model of PTSD in mice. The morris water maze test, open field test and contextual fear test were conducted to assess memory deficits, anxiety-like behaviors, and freezing of mice. Western blot was performed to identify proteins and reveal their levels in hippocampal tissues. We found that mice receiving SPS exhibited increased anxiety-like behaviors, memory deficits, and prolonged freezing time. The protein levels of BHLHE40 were downregulated in the hippocampal tissues of SPS mice. SPS reduced the protein levels of glutamate receptors, while overexpression of BHLHE40 promoted glutamate receptor protein levels in SPS mice. Moreover, BHLHE40 overexpression activated the PI3K/AKT pathway. BHLHE40 overexpression ameliorated the SPS-induced PTSD-like behavioral deficits. Overall, BHLHE40 promotes glutamate receptor protein levels to ameliorate PTSD-like behaviors with the involvement of the PI3K/AKT pathway. This novel discovery may provide a potential target for the improvement of PTSD.
To study the effects of delayed resuscitation on the apoptosis rate (ap%) and expression of apoptosis-related genes of lamina propria lymphocytes (LPL), intra-epithelial lymphocyte(IEL) after burn in rats.Thirty Wistar rats were randomly divided into early resuscitation (ER) and delayed resuscitation (DR) groups, 24 rats of them were subjected to 30% total body surface area (TBSA) III degree scald burn on back, among which 12 received fluid resuscitation immediately, and 12 with delayed fluid resuscitation. Six rats were subjected to sham burn served as control. DNA fragmentation, DNA agarose gel electrophoresis were used to observe the apoptosis of LPL and IEL. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to detect the expression of interleukin-1beta-converting enzyme (ICE) and B cell lymphoma-2 (bcl-2).The ap% of LPL and IEL increased significantly after burn (all P<0.01). The ap% in DR group was higher than in ER group at 12 hours postburn. Distinct DNA "ladders" were visualized in agarose gel electrophoresis 6 hours postburn for both LPL and IEL. The expression of ICE gene increased dramatically after burn, and it expressed higher in DR group than that in ER group postburn (P<0.05 or P<0.01). A strong expression of bcl-2 in LPL and IEL were detected before burn, but the expressions decreased significantly after burn and delayed resuscitation (P<0.05 or P<0.01).The apoptosis of LPL and IEL increased significantly after burn and delayed resuscitation. The unbalanced expression of apoptosis-enhanced gene ICE and apoptosis-inhibited gene bcl-2 may be responsible for the dramatically increased apoptosis of LPL and IEL postburn.
Background Visfatin is an adipose-derived adipocytokine involved in nicotinamide adenine dinucleotide (NAD) synthesis.The expression of visfatin was affected by the obesity,distribution of adipose tissue,and the inflammatory state,etc.However,visfatin level in hypertension,diabetes,and atherosclerosis was controversial.Objective To investigate the expression of the adiponectin and visfatin in adipose tissues and its relationship with components of metabolic syndrome (MS).Methods Thirty-six MS patients and 32 age and sex matched non-MS control subjects underwent operation due to gallstone were enrolled.Data on height,body weight,waist circumference (WC) and hip circumference (HC) were collected.Fasting plasma glucose (FPG),triglyceride (TG),total cholesterol (TC),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol(LDL-C),fasting insulin,adiponectin and visfatin were determined.Expressions of adiponectin and visfatin in abdominal subcutaneous and omental adipose tissues were assessed by semi-quantitive RT-PCR.Results The adiponectin mRNA expression in omental adipose tissue was significantly lower in MS patients than that in control subjects (0.73±0.16 vs 0.83±0.17,P0.05).In male subjects,the level of adiponectin mRNA expression in omental adipose tissue was lower in MS group than that in non-MS group (0.69±0.17 vs 0.83±0.19,P0.05).However,there is no significant difference of visfatin mRNA expression between two groups.Partial correlation analysis showed that the level of adiponectin in blood plasma was positively correlated with HDL-C (r=0.513,P0.05).The expression of visfatin mRNA in omental adipose tissue was positively correlated with HOMA-IR (r=0.219,P0.05),and the level of visfatin in blood plasma was positively correlated with TC,TG and LDL-C.Conclusion Adiponectin level in visceral fat tissue may be related with metabolic syndrome,while no differences in visfatin were found between patients with and without metabolic syndrome.
Objective To investigate the effect of rhPTH (1-34) and elcatonin on bone metabolism and serum secreted protein acidic and rich in cysteine ( SPARC ) in postmenopausal women with osteoporosis.Methods One hundred and twenty-four postmenopausal women with osteoporosis were randomly divided into 2 groups:One group was treated with recombinant human parathyroid hormone ( 1-34 ) [ rhPTH ( 1-34 ) ] 200 U/d by subcutaneous injection (PTH group,n =89 )and another group was treated with elcatonin 20 U/week by intramuscular injection (CT group,n =35 ) for 12 months.All patients received a basic therapy with oral calcium ( Ca 600 mg+ Vit D3125 U,q..d.).The bone mineral density ( BMD ) of lumbar spine( L2-4 ),the left femoral neck,greater trochanter,and Ward's triangle,serum calcium and phosphate were measured by baseline,6 months' and 12 months.Levels of serum bone-specific alkaline phosphatase( BSAP),serum secreted protein acidic and rich in cysteine (SPARC)were determined by an ELISA assay.Results By 12 months,rhPTH ( 1-34 ) treatment significantly increased the lumbar spine L2-4 BMD 7.9％ (P＜0.05),serum calcium 8.3 ％ ( P＜ 0.05 ),serum BSAP 93.4％ ( P＜ 0.05 ),serum SPARC by 12.6％[ ( 195.68±59.57 vs 173.81 ±81.33 ) pμg/L,P＜0.05 ].Elcatonin therapy increased the lumbar spine L2-4 BMD by 3.2％ (P＜0.05) at the end of 12 months,but elcatonin did not influence serum calcium,BSAP and SPARC.The rhPTH( 1-34 ) increased lumbar spine L2-4 BMD more than elcatonin did at 12 months( P＜0.05 ).Conclusion rhPTH (1-34) could promote the bone anabolism more effectively than elcatonin did.Serum SPARC may play an important role in promoting osteogenesis by rhPTH.
Key words:
Recombinant human parathyroid hormone (1-34); Elcatonin; Osteoporosis; Secreted protein acidic and rich in cysteine
To construct the recombinant plasmid containing human microdystrophin cDNA, and study the microdystrophin expression in vivo and in vitro.Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I, the product was inserted into plasmid pVAX1, resulting in pAMICDYS. And then 3T3 cells were transfected with pAMICDYS. Forty-eight hours after transfection, the expression of the microdystrophin was detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Finally, TA muscles of mdx mice were injected with the recombinant plasmid pAMICDYS through i.m. and the pathological change of TA was evaluated by histology, and the expression of microdystrophin in mdx TA was detected by immunohistochemical analysis.The recombinant plasmid containing human microdystrophin cDNA was constructed successfully. The recombinant plasmid was proved to be able to express microdystrophin protein both in vivo and in vitro. Moreover, treatment of the TA of mdx mice with the recombinant plasmid could decrease the number of centrally nucleated myofibers.Recombinant plasmid containing the microdystrophin gene was constructed successfully, and it could express microdystrophin protein both in vivo and in vitro. It provides basis for further study on microdystrophin as a target gene to treat Duchenne muscular dystrophy (DMD) by electrotransfer, i.v, arterial injection and combining with other exogenous gene to enhance microdystrophin expression.
Deeper studies on the pathological mechanism associated with invasiveness of non-functioning pituitary adenoma (NFPA) is imperative to find better treatments. This research was preliminarily conducted to investigate the correlation between the expression of Claudin-9 (CLDN9), Tyrosine kinase-2 (TYK2), Signal transducers and activators of transcription-3 (STAT3) and invasiveness in NFPA to illustrate the pathological mechanism.
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