Analysis of the functional role and mRNA expression of GABABR in the nucleus accumbens of cocaine-addicted rats.

BACKGROUND: Drug addiction is a social and medical problem that must be urgently addressed. The nucleus accumbens (NAc) is closely related to addiction-related learning memory, and γ-aminobutyric acid type B receptor (GABABR) is a potential target for the treatment of drug addiction. However, the role of GABABR activity levels in the NAc in cocaine addiction is unclear. METHODS: In this study, we established an animal model of cocaine dependence, modulated the level of GABABR activity, applied a conditioned place preference assay (CPP) to assess the role of the NAc in reconsolidation of addiction memory, evaluated learning and memory functions by behavioral experiments, examined the expression of GB1, GB2, CREB, p-CREB, PKA, ERK, and BDNF in the NAc by molecular biology experiments, and screened differentially significantly expressed genes by transcriptome sequencing. RESULTS: Our study showed that the GABAB receptor agonist BLF had a significant effect on locomotor distance in rats, promoted an increase in GABA levels and significantly inhibited the PKA and ERK1/2/CREB/BDNF signaling pathways. Moreover, transcriptome sequencing showed that GABABR antagonist intervention identified a total of 21 upregulated mRNAs and 21 downregulated mRNAs. The DE mRNA genes were mainly enriched in tyrosine metabolism; however, further study is needed. CONCLUSION: GABABR activity in the NAc is involved in the regulation of cocaine addiction and may play an important role through key mRNA pathways.

Association between microRNA 21 expression in serum and lung cancer: A protocol of systematic review and meta-analysis.

BACKGROUND: Previous studies have reported that microRNA 21 (mRNA 21) has involved in the procedure of lung cancer (LC). However, its conclusions are still unclear. Thus, this study will try to elaborate the association between mRNA 21 expression in serum and LC. METHODS: The electronic databases of Cochrane Library, PubMed, EMBASE, Allied and Complementary Medicine Database, WANGFANG database, and China National Knowledge Infrastructure will be retrieved from the inception to the present. All electronic databases will be searched without limitations of language and geographical location. Case-controlled studies reporting the association between mRNA 21 expression in serum and LC will be included. In addition, we will also identify other literature sources to avoid missing potential studies. All study selection, information collection, and study quality assessment will be performed by 2 independent authors. RevMan V.5.3 software and Stata V.12.0 software will be used for data synthesis and analysis. RESULTS: This study will summarize current evidence to investigate the association between mRNA 21 expression in serum and LC. CONCLUSION: The findings of this study will present comprehensive evidence to determine whether mRNA 21 expression in serum is relevant with LC or not. SYSTEMATIC REVIEW REGISTRATION: INPLASY202040055.

MicroRNA-370 protects against myocardial ischemia/reperfusion injury in mice following sevoflurane anesthetic preconditioning through PLIN5-dependent PPAR signaling pathway.

Myocardial ischemia/reperfusion injury (IRI) has long been identified to be a contributor to adverse cardiovascular outcomes following myocardial ischemia, cardiac surgery or circulatory arrest. This study aims to investigate the effects of microRNA (miR-370) targeting perilipin-5 (PLIN5) in mice following sevoflurane anesthetic preconditioning (SAP). A mouse model of left ventricular myocardial IRI was established, followed by the evaluation of myocardial infarction size and cardiac function to determine the effects of SAP. The underlying regulatory mechanisms of miR-370 were analyzed in concert with the treatment of miR-370 mimic, miR-370 inhibitor, or siRNA against PLIN5 in cardiomyocytes isolated from mice with IRI. Also, cardiomyocyte proliferation, cell cycle distribution and apoptosis were evaluated following treatment. Lastly, SAP-treated I/R mice were injected with miR-370 inhibitor to verify the mechanism of SAP. The use of SAP conferred cardioprotective effects on myocardial IRI. MiR-370 was downregulated in mice that exhibited IRI, but SAP elevated the miR-370 expression. Functionally, miR-370 negatively targeted PLIN5 and activated the peroxisome proliferator activated-receptor (PPAR) signaling pathway, leading to decreased PPARγ expression but increased PPARα expression. The results also showed that elevation of miR-370 or the silencing of PLIN5 promoted cardiomyocyte proliferation. miR-370 also inhibited cardiomyocyte apoptosis as reflected by decreased caspase-3 expression and increased Bcl-2 expression. Additionally, SAP also alleviated I/R injury by inhibiting PPARγ. This study demonstrates that SAP induces miR-370 and exerts cardioprotective effects on myocardial IRI, where upregulation of miR-370 alleviates myocardial IRI via inhibiting the PLIN5-dependent PPAR signaling pathway.

FAT10 is associated with the malignancy and drug resistance of non-small-cell lung cancer.

Lung cancer has become one of the leading causes of cancer mortality worldwide, and non-small-cell lung cancer (NSCLC) accounts for ~85% of all lung cancer cases. Currently, platinum-based chemotherapy drugs, including cisplatin and carboplatin, are the most effective treatment for NSCLC. However, the clinical efficacy of chemotherapy is markedly reduced later in the treatment because drug resistance develops during the treatment. Recently, a series of studies has suggested the involvement of FAT10 in the development and malignancy of multiple cancer types. In this study, we focused our research on the function of FAT10 in NSCLC, which has not been previously reported in the literature. We found that the expression levels of FAT10 were elevated in quick chemoresistance NSCLC tissues, and we demonstrated that FAT10 promotes NSCLC cell proliferation, migration, and invasion. Furthermore, the protein levels of FAT10 were elevated in cisplatin- and carboplatin-resistant NSCLC cells, and knockdown of FAT10 reduced the drug resistance of NSCLC cells. In addition, we gained evidence that FAT10 regulates NSCLC malignancy and drug resistance by modulating the activity of the nuclear factor kappa B signaling pathway.

Navigating into the binding pockets of the HER family protein kinases: discovery of novel EGFR inhibitor as antitumor agent.

The epidermal growth factor receptor (EGFR) family has been validated as a successful antitumor drug target for decades. Known EGFR inhibitors were exposed to distinct drug resistance against the various EGFR mutants within non-small-cell lung cancer (NSCLC), particularly the T790M mutation. Although so far a number of studies have been reported on the development of third-generation EGFR inhibitors for overcoming the resistance issue, the design procedure largely depends on the intuition of medicinal chemists. Here we retrospectively make a detailed analysis of the 42 EGFR family protein crystal complexes deposited in the Protein Data Bank (PDB). Based on the analysis of inhibitor binding modes in the kinase catalytic cleft, we identified a potent EGFR inhibitor (compound A-10) against drug-resistant EGFR through fragment-based drug design. This compound showed at least 30-fold more potency against EGFR T790M than the two control molecules erlotinib and gefitinib in vitro. Moreover, it could exhibit potent HER2 inhibitory activities as well as tumor growth inhibitory activity. Molecular docking studies revealed a structural basis for the increased potency and mutant selectivity of this compound. Compound A-10 may be selected as a promising candidate in further preclinical studies. In addition, our findings could provide a powerful strategy to identify novel selective kinase inhibitors on the basis of detailed kinase-ligand interaction space in the PDB.

Biomechanical effects of cervical arthroplasty with U-shaped disc implant on segmental range of motion and loading of surrounding soft tissue.

PURPOSE: Various design concepts have been adopted in cervical disc prostheses, including sliding articulation and standalone configuration. This study aimed to evaluate the biomechanical effects of the standalone U-shaped configuration on the cervical spine. METHODS: Based on an intact finite element model of C3-C7, a standalone U-shaped implant (DCI) was installed at C5-C6 and compared with a sliding articulation design (Prodisc-C) and an anterior fusion system. The range of motion (ROM), adjacent intradiscal pressure (IDP) and capsular ligament strain were calculated under different spinal motions. RESULTS: Compared to the intact configuration, the ROM at C5-C6 was reduced by 90% after fusion, but increased by 70% in the Prodisc-C model, while the maximum percentage change in the DCI model was 30% decrease. At the adjacent segments, up to 32% increase in ROM happened after fusion, while up to 34% decrease occurred in Prodisc-C model and 17% decrease in DCI model. The IDP increased by 11.6% after fusion, but decreased by 5.6 and 6.3% in the DCI and Prodisc-C model, respectively. The capsular ligament strain increased by 147% in Prodisc-C and by 13% in the DCI model. The DCI implant exhibited a high stress distribution. CONCLUSIONS: Spinal fusion resulted in compensatory increase of ROM at the adjacent sites, thereby elevating the IDP. Prodisc-C resulted in hyper-mobility at the operative site that led to an increase of ligament force and strain. The U-shaped implant could maintain the spinal kinematics and impose minimum influence on the adjacent soft tissues, despite the standalone configuration encountering the disadvantages of high stress distribution.

Cervical disc arthroplasty with ProDisc-C artificial disc: 5-year radiographic follow-up results.

BACKGROUND: Cervical disc arthroplasty is an alternative surgery to standard cervical decompression and fusion for disc degeneration. Different types of cervical disc prosthesis are used in China. The aim of this study was to evaluate the radiographic outcomes of cervical arthroplasty using the ProDisc-C prosthesis. METHODS: Radiographic evaluation, including static and dynamic flexion-extension lateral images, was performed at baseline and at final follow-up. RESULTS: Twenty six patients who had single-level ProDisc-C arthroplasty were followed up for a mean period of 63 months (56-76 months). The range of motion at the operated level was 9.3°±3.7° at baseline and 7.3°±3.5° at final follow-up, with a significant difference (P < 0.05). Seventeen of 26 levels (65.4%) developed heterotopic ossification: three were classified as grade II, 13 were classified as grade III, and 1 as grade IV, according to McAfee's classification. Forty nine adjacent segments were evaluated by lateral X-ray and 18 (36.7%) segments developed adjacent segment degenerations. CONCLUSIONS: ProDisc-C arthroplasty had acceptable radiographic results at 5-year follow-up. The range of motion was preserved. However, more than 60% of the patients developed heterotopic ossification.

Comparison of adjacent segment degeneration five years after single level cervical fusion and cervical arthroplasty: a retrospective controlled study.

BACKGROUND: Cervical arthroplasty is indicated to preserve cervical motion and prevent accelerated adjacent segment degeneration. Whether accelerated adjacent segment degeneration is prevented in the long term is unclear. This trial compared adjacent segment degeneration in Bryan disc arthroplasty with that in anterior cervical decompression and fusion five years after the surgery. METHODS: We studied patients with single level degenerative cervical disc disease. The extent of adjacent segment degeneration was estimated from lateral X-rays. RESULTS: Twenty-six patients underwent single level Bryan disc arthroplasty and twenty-four patients underwent single level anterior cervical decompression and fusion. All patients were followed up for an average of sixty months. In the Bryan arthroplasty group, nine (17.6%) segments developed adjacent segment degeneration, which was significantly lower than that (60.4%) in the anterior cervical decompression and fusion group. Eleven segments in the Bryan arthroplasty group developed heterotopic ossification according to McAfee's classification and two segments had range of motion less than 2°. In the heterotopic ossification group, four (19.5%) segments developed adjacent segment degeneration, similar to the number in the non-heterotopic ossification group (16.7%). Adjacent segment degeneration rate was 50% in grade IV group but 11.8% in grade II to III. CONCLUSIONS: Adjacent segment degeneration was accelerated after anterior cervical decompression and fusion. However, Bryan disc arthroplasty avoided accelerated adjacent segment degeneration by preserving motion. Patients with grade IV heterotopic ossification lost motion, and the rate of adjacent segment degeneration was higher than that in patients without heterotopic ossification.

The extraction of LRP via functional data analysis techniques.

A new strategy based on functional data analysis (FDA) techniques is proposed to extract the lateralized readiness potential (LRP), which treats electroencephalographic data as functional data. This FDA-based method combines longitudinal information from each trial (time series data) with cross-sectional information from all trials at a fixed time point (cross-sectional data). The comparison results show that the FDA-based LRP is closer to the assumed true LRP and is more robust against a reduction in the number of trials than the traditional average-based LRP. Furthermore, the results indicate that the onset of an FDA-based LRP is more accurate than that of an average-based LRP under several measuring criteria.

Application of cervical arthroplasty with Bryan cervical disc: long-term X-ray and magnetic resonance imaging follow-up results.

BACKGROUND: Cervical disc arthroplasty is a new technique for treating degenerative cervical disease. Its goal is to avoid the degeneration of adjacent levels by preserving motion at the treated level. The aims of this study were to evaluate the radiologic outcomes of Bryan cervical disc replacement and the degenerative status of adjacent segments. METHODS: Twenty-two patients at a single center underwent discectomy and implantation of Bryan cervical disc. The mean follow-up period was 60 months (57 - 69 months). Twenty patients underwent single-level arthroplasty and two underwent arthroplasty at two levels. The levels of surgery included C3/4 (3 levels), C4/5 (2 levels), C5/6 (18 levels) and C6/7 (1 level). Radiographic evaluation included dynamic X-ray examination and magnetic resonance imaging (MRI) at baseline and at final follow-up. RESULTS: On X-ray examination, the range of motion (ROM) at the operated level was 7.2° (2.5° - 13.0°) at baseline and 7.8° (1.0° - 15.0°) at final follow-up (P > 0.05). Heterotopic ossification around the prosthesis was observed in eight levels, and two levels showed loss of motion (ROM < 2°). MRI showed worsening by a grade at the upper level in 2/22 patients, and worsening by a grade at the lower level in 3/22, according to Miyazaki's classification. No further impingement of the ligamentum flavum into the spinal canal was observed at adjacent levels, though the disc bulge was slightly increased at both the adjacent upper and lower levels at final follow-up. CONCLUSIONS: Arthroplasty using Bryan cervical disc prosthesis resulted in favorable radiologic outcomes in this study. Disc degeneration at adjacent levels may be postponed by this technique.

[The application of cervical spine arthroplasty with Bryan disc and mid-term clinical and radiological follow-up results].

OBJECTIVE: To study the clinical and radiological outcome of Bryan cervical disc replacement and the degenerative status on adjacent segments. METHODS: The data of 26 cases of single level disc replacement with minimal 2 years follow-up were reviewed. CLINICAL OUTCOME was assessed with the JOA 17 score scale and Odom's score. Radiological assessment including range of motion and heterotopic ossification of operated level were recorded. Adjacent level degeneration on X-ray and MRI scan at baseline and at follow-up were compared. RESULTS: (1) CLINICAL OUTCOME: the average JOA score was 16 with 84% improvement ratio at final follow-up in 18 cases of cervical myelopathy. Eight cases of radiculopathy were fully recovered. According to the Odom's criteria 15 cases had an excellent outcome, 7 good, 4 fair, and no case of poor result. (2) On X-ray: The range of motion (ROM) at operated level was 6.9 degrees (2 degrees-12 degrees) at baseline and 7.8 degrees (1 degree-14 degrees) at final follow-up. The heterotopic ossification around the prosthesis was observed in 7 cases and only 1 case lost movement. The average ROM was 5.3 degree in other 6 cases. There was no obvious change of disc height at adjacent levels. (3) On MRI: There was no deterioration of disc degeneration at adjacent levels at final follow-up according to Pfirrmann's classification. There was no further ligamentum flavum impingement into spinal canal observed at adjacent levels but the disc movement slightly increased at both upper and lower adjacent level at final follow-up. CONCLUSIONS: The motion at operated level is preserved after minimal 2 years Bryan disc replacement with satisfied clinical outcome. The deterioration of disc degeneration at adjacent levels may be postponed.

[Gene cloning, sequence analysis and tissue expression of estrogen-related receptor alpha (Erralpha) in Japanese medaka and its transcriptional responses after differential EDCs exposure].

Endocrine disrupting chemicals (EDCs) can bind or block nuclear receptors in the body and subsequently affect growth, development and reproduction of fish. Estrogen-related receptors (ERRs), belonging to the nuclear receptor superfamily, have been implicated in diverse physiological processes in estrogen signal pathway in mammals, while little is known about them in fishes. Complete mRNA sequence of ERRalpha from medaka (Oryzias latipes) was cloned, and the sequence is similar to those of other vertebrates, especially that the DNA-binding domain (DBD) of ERRalpha is highly conserved among the vertebrates (97.4%-100% sequence identities) and the ligand-binding domain (LBD) of medaka ERRalpha is 66.4%-67.0% sequence identities with those of mammals. The DBD of medaka ERRalpha is of the same length and has high sequence identity with those of estrogen receptor (ERalpha and ERbeta) and androgen receptor (ARalpha and ARbeta) of medaka, but much difference was found between the LBD of medaka ERRalpha with those of ERalpha, ERbeta, ARalpha and ARbeta. ERRalpha gene is located in chromosome 14 and is consisted of 5 exons. The expressions of ERRalpha in different tissues and the transcriptional responses of ERRalpha in testis of medaka exposed differential EDCs were studied by quantitative real-time RT-PCR. ERRalpha is expressed at apparently high levels in gonad, brain, eye, spleen and intestine, though it was broadly expressed in tissues. Significant transcriptional difference was found between testis and ovary, implying ERRalpha would be involved in sex differentiation and gonad development in fish. After 3 weeks exposure of medaka to 200 ng/L ethynylestradiol (EE2), 200 ng/L estrone (E1), 200 ng/L diethylstilbestrol (DES), 100 microg/L atrazine (AT) and 200 ng/L 17beta-estradiol (E2), transcripts of ERRalpha were significantly decreased to 0.54, 0.56, 0.61, 0.63 and 0.65 of control (p < 0.05) in the testes, respectively. And those in the 1 microg/L tributyltin (TBT) and 1 microg/L triphenyltin (TPT) exposure groups were up-regulated to 1.34 and 1.35 folds of control (p > 0.05), respectively. These results suggested that ERRalpha would take actions in the disruption of sex differentiation and gonad development in fish by EDCs. In addition, no multiple steroid hormone-response element half-sites was found in medaka, which were reported in the upstream of ERRalpha gene in mammals, indicating there would be different regulation patters of ERRalpha between teleost and mammal.

Clinical and dosimetric factors of radiation-induced esophageal injury: radiation-induced esophageal toxicity.

AIM: To analyze the clinical and dosimetric predictive factors for radiation-induced esophageal injury in patients with non-small-cell lung cancer (NSCLC) during three-dimensional conformal radiotherapy (3D-CRT). METHODS: We retrospectively analyzed 208 consecutive patients (146 men and 62 women) with NSCLC treated with 3D-CRT. The median age of the patients was 64 years (range 35-87 years). The clinical and treatment parameters including gender, age, performance status, sequential chemotherapy, concurrent chemotherapy, presence of carinal or subcarinal lymph nodes, pretreatment weight loss, mean dose to the entire esophagus, maximal point dose to the esophagus, and percentage of volume of esophagus receiving >55 Gy were studied. Clinical and dosimetric factors for radiation-induced acute and late grade 3-5 esophageal injury were analyzed according to Radiation Therapy Oncology Group (RTOG) criteria. RESULTS: Twenty-five (12%) of the two hundred and eight patients developed acute or late grade 3-5 esophageal injury. Among them, nine patients had both acute and late grade 3-5 esophageal injury, two died of late esophageal perforation. Concurrent chemotherapy and maximal point dose to the esophagus > or =60 Gy were significantly associated with the risk of grade 3-5 esophageal injury. Fifty-four (26%) of the two hundred and eight patients received concurrent chemotherapy. Among them, 25 (46%) developed grade 3-5 esophageal injury (P = 0.0001<0.01). However, no grade 3-5 esophageal injury occurred in patients who received a maximal point dose to the esophagus <60 Gy (P = 0.0001<0.01). CONCLUSION: Concurrent chemotherapy and the maximal esophageal point dose > or =60 Gy are significantly associated with the risk of grade 3-5 esophageal injury in patients with NSCLC treated with 3D-CRT.