The Role of Paraspinal Muscle in Postoperative Coronal Balance Transition in Degenerative Lumbar Scoliosis: A 2-year Follow-up Study.

STUDY DESIGN: Retrospective observational study. OBJECTIVE: The purpose of this study was to determine whether paraspinal muscle could influence postoperative coronal balance and its transition in degenerative lumbar scoliosis (DLS). SUMMARY OF BACKGROUND DATA: Although the importance of the paraspinal muscles (PSM) in sagittal alignment is well recognized, there is no information about its role in coronal balance. METHODS: The study included 102 DLS patients. Evaluation of the PSM on magnetic resonance imaging were conducted at baseline. Coronal measurements included coronal balance distance (CBD), major Cobb angle, L4 coronal tilt, and L5 coronal tilt. The cohort was divided based on postoperative parameters into persistent coronal balance (PCB), worsened coronal imbalance (WCIB), recurrent coronal balance (RCB), and persistent coronal imbalance (PCIB) according to immediate postoperative and follow-up coronal balance. Multivariate logistic regression models for postoperative CIB, follow-up WCIB and follow-up RCB were utilized to identify statistically significant associations while accounting for confounders. RESULTS: The cohort was divided into 57 with PCB, 13 with WCIB, 10 with RCB, and 22 with PCIB. The follow-up groups with CIB exhibited more severe fatty infiltration in the extensor muscle compared to the balanced groups. Specifically, the WCIB group demonstrated the most severe extensor muscle degeneration, particularly on the concave sides, and the most prominent asymmetrical degeneration of the PSM among the four groups. Furthermore, patients with CIB had worse sagittal malalignment compared to those with CB at the last follow up. CONCLUSION: Patients exhibiting stronger extensor muscle mass were prone to immediate postoperative CB and more likely to experience spontaneous improvement or recurrence of coronal balance during follow-up. Severe extensor muscle degeneration and prominent asymmetrical bilateral PSM degeneration represent potential risk factors for persistent CIB and recurrent CIB. It is crucial to assess the dynamic change during the follow-up period as long-term prognosis may be impacted if CB deteriorates, or otherwise develops during follow-up. LEVEL OF EVIDENCE: 3.

The establishment of a novel upper cervical complex fracture classification system.

BACKGROUND CONTEXT: Upper cervical complex fractures are associated with high rates of neurological damage and mortality. The Dickman's classification is widely used in the diagnosis of upper cervical complex fractures. However, it falls short of covering the full spectrum of complex fractures. This limitation hinders effective diagnosis and treatment of these injuries. PURPOSE: To address the diagnostic gap in upper cervical complex fractures, the study introduces a novel classification system for these injuries, assessing its reliability and usability. STUDY DESIGN: Proposal of a new classification system for upper cervical complex fractures. PATIENT SAMPLE: The study comprised the clinical data of 242 patients with upper cervical complex fractures, including 32 patients treated at our hospital, along with an additional 210 cases from the literature. OUTCOME MEASURES: The inter-observer and intra-observer reliability (kappa coefficient, κ) of this classification system were investigated by 3 spine surgeons. The 3 researchers independently re-evaluated the upper cervical complex fracture classification system 3 months later. METHODS: The proposed classification categorizes upper cervical complex fractures into 3 main types: Type I combines odontoid and Hangman's fractures into 2 subtypes; Type II merges C1 with odontoid/Hangman's fractures into 3 subtypes; and Type III encompasses a combination of C1, odontoid, and Hangman's fractures, divided into 2 subtypes. Meanwhile, a questionnaire was administered in 15 assessors to evaluate the system's ease of use and clinical applicability. RESULTS: A total of 45 cases (18.6%) unclassifiable by Dickman's classification were successfully categorized using our system. The mean κ value of inter-observer reliability was 0.783, indicating substantial reliability. The mean κ value of intra-observer reliability was 0.862, indicating almost perfect reliability. Meanwhile, thirteen assessors (87.7%) stated that the classification system is easy to remember, easy to apply, and they expressed intentions to apply it in clinical practice in the future. CONCLUSIONS: This system not only offers high confidence and reproducibility but also serves as a precise guide for clinicians in formulating treatment plans. Future prospective applications are warranted to further evaluate this classification system.

Can the 6-minute Walking Test Assess Ambulatory Function Impairment in Patients with Cervical Spondylotic Myelopathy?

STUDY DESIGN: Prospective cohort study. OBJECTIVE: Investigating the ability of a 6-minute walking test (6MWT) to assess functional status in patients with cervical spondylotic myelopathy (CSM). SUMMARY OF BACKGROUND DATA: The 6MWT provides an objective assessment of a patient's ability to walk. There is the potential for its application to the assessment of functional status in patients with CSM. MATERIALS AND METHODS: One hundred and thirty-five patients from our institution were prospectively enrolled from July 2022 to August 2023. A control group of age- and sex-matched healthy individuals was established. The 6MWT was conducted in strict accordance with established guidelines. The Nurick score, the Prolo score, the Cooper-myelopathy-scale score (CMS), the Japanese Orthopedic Association score (JOA) and the European-myelopathy-scale score (EMS) were assessed preoperatively. Visual Analog Scale (VAS) for pain or numbness and Oswestry Neck Disability Index (NDI) were also collected. Radiographic parameters were measured and recorded. Continuous variables between patients and controls were compared by applying the t-test. The chi-square test was used to compare gender ratios between groups. Pearson's correlation analysis was used to analyze the association between continuous variables and ordinal variables. Subgroups of CSM patients were analyzed according to global spinal alignment types, based on whether the SVA was greater than or equal to 50 mm. Clinical scores and imaging parameters were compared by t-test. RESULTS: The preoperative 6-minute walking distance (6MWD) of CSM patients was 309.34 ± 116.71 m, which was significantly lower than that of the controls (464.30 ± 52.59 m, P<.01). The 6MWD was significantly correlated with scores on all clinical scales except the VAS. CMS Lower extremity score had the strongest correlation with preoperative 6MWD in CSM patients (r=-.794 , P<.01). Of the sagittal alignment parameters, only C7 sagittal vertical axis (SVA) and T1 slope were significantly correlated with 6MWD(r=-.510,-0.360, respectively). CSM patients with SVA greater than 50 mm had significantly lower 6MWD than CSM patients with SVA less than or equal to 50 mm (168.00 ± 137.26 vs 346.24 ± 84.27 m, P<.01). CONCLUSIONS: The 6MWD of CSM patients was significantly lower than that of the healthy population and correlated well with commonly used clinical scales. The 6MWD can potentially assist in the assessment of functional status in patients with CSM.

Piezo1 channel exaggerates ferroptosis of nucleus pulposus cells by mediating mechanical stress-induced iron influx.

To date, several molecules have been found to facilitate iron influx, while the types of iron influx channels remain to be elucidated. Here, Piezo1 channel was identified as a key iron transporter in response to mechanical stress. Piezo1-mediated iron overload disturbed iron metabolism and exaggerated ferroptosis in nucleus pulposus cells (NPCs). Importantly, Piezo1-induced iron influx was independent of the transferrin receptor (TFRC), a well-recognized iron gatekeeper. Furthermore, pharmacological inactivation of Piezo1 profoundly reduced iron accumulation, alleviated mitochondrial ROS, and suppressed ferroptotic alterations in stimulation of mechanical stress. Moreover, conditional knockout of Piezo1 (Col2a1-CreERT Piezo1flox/flox) attenuated the mechanical injury-induced intervertebral disc degeneration (IVDD). Notably, the protective effect of Piezo1 deficiency in IVDD was dampened in Piezo1/Gpx4 conditional double knockout (cDKO) mice (Col2a1-CreERT Piezo1flox/flox/Gpx4flox/flox). These findings suggest that Piezo1 is a potential determinant of iron influx, indicating that the Piezo1-iron-ferroptosis axis might shed light on the treatment of mechanical stress-induced diseases.

Understanding the Impacts of Surface Compositions on the In-Vitro Dissolution and Aerosolization of Co-Spray-Dried Composite Powder Formulations for Inhalation.

PURPOSE: Dissolution behavior of dry powder inhaler (DPI) antibiotic formulations in the airways may affect their efficacy especially for poorly-soluble antibiotics such as azithromycin. The main objective of this study was to understand the effects of surface composition on the dissolution of spray dried azithromycin powders by itself and in combination with colistin. METHODS: Composite formulations of azithromycin (a poorly water-soluble molecule) and colistin (a water-soluble molecule) were produced by spray drying. The resultant formulations were characterized for particle size, morphology, surface composition, solid-state properties, solubility and dissolution. RESULTS: The results demonstrate that surfaces composition has critical impacts on the dissolution of composite formulations. Colistin was shown to increase the solubility of azithromycin. For composite formulations with no surface colistin, azithromycin released at a similar dissolution rate as the spray-dried azithromycin alone. An increase in surface colistin concentration was shown to accelerate the dissolution of azithromycin. The dissolution of colistin from the composite formulations was significantly slower than the spray-dried pure colistin. In addition, FTIR spectrum showed intermolecular interactions between azithromycin and colistin in the composite formulations, which could contribute to the enhanced solubility and dissolution of azithromycin. CONCLUSIONS: Our study provides fundamental understanding of the effects of surface concentration of colistin on azithromycin dissolution of co-spray-dried composite powder formulations.

Applications of thin film plastic scintillator in measurement of soft x rays generated from Z-pinch implosion.

A thin film plastic scintillator detector has been developed for the measurement of radiation power and yield of soft x rays produced from Z-pinch implosion. To enable soft x-ray measurements using plastic scintillators, the detector geometry has been specially designed to minimize visible light and alleviate nonlinear behavior. Energy response has been calibrated, and saturation effects have been explored and described in details. The possibility and limitation of its application to such high-density radiation bursts are analyzed. The detector has been fielded on several meters away in vacuum pipes for hundreds of shots at different Z-pinch facilities, and the measured data in these experiments agreed well with the results from other diagnostics, demonstrating the feasibility and reliability of the detector.

Physico-Chemical Properties, Aerosolization and Dissolution of Co-Spray Dried Azithromycin Particles with L-Leucine for Inhalation.

PURPOSE: Inhalation therapy is popular to treat lower respiratory tract infections. Azithromycin is effective against some bacteria that cause respiratory tract infections; but it has poor water solubility that may limit its efficacy when administrated as inhalation therapy. In this study, dry powder inhaler formulations were developed by co-spray drying azithromycin with L-leucine with a purpose to improve dissolution. METHODS: The produced powder formulations were characterized regarding particle size, morphology, surface composition and in-vitro aerosolization performance. Effects of L-leucine on the solubility and in-vitro dissolution of azithromycin were also evaluated. RESULTS: The spray dried azithromycin alone formulation exhibited a satisfactory aerosol performance with a fine particle fraction (FPF) of 62.5 ± 4.1%. Addition of L-leucine in the formulation resulted in no significant change in particle morphology and FPF, which can be attributed to enrichment of azithromycin on the surfaces of composite particles. Importantly, compared with the spray-dried amorphous azithromycin alone powder, the co-spray dried powder formulations of azithromycin and L-leucine demonstrated a substantially enhanced in-vitro dissolution rate. Such enhanced dissolution of azithromycin could be attributed to the formation of composite system and the acidic microenvironment around azithromycin molecules created by the dissolution of acidic L-leucine in the co-spray dried powder. Fourier transform infrared spectroscopic data showed intermolecular interactions between azithromycin and L-leucine in the co-spray dried formulations. CONCLUSIONS: We developed the dry powder formulations with satisfactory aerosol performance and enhanced dissolution for a poorly water soluble weak base, azithromycin, by co-spray drying with an amino acid, L-leucine.

Structure and extreme ultraviolet performance of Si/C multilayers deposited under different working pressures.

Narrow bandwidth Si/C multilayer mirrors are fabricated and characterized for the Z-pinch plasma diagnostic at a wavelength of 16.5 nm. To reduce the large stress of the multilayer and maintain a practical reflectivity, different working pressures, from 0.13 Pa to 0.52 Pa, are optimized during the deposition. The grazing incidence x-ray reflectometry (GIXR) measurement and the fitting results indicate that an interlayer was formed at the interfaces, while both the interlayer thickness and interface widths increase with larger working pressure. The surface roughness of the multilayers also increases from 0.13 nm at 0.13 Pa to 0.29 nm at 0.52 Pa, as revealed by the atomic force microscope (AFM) measurements. The multilayer stress decreases from -682 MPa to -384 MPa as the working pressure increases from 0.13 Pa to 0.52 Pa, respectively. The experimental extreme ultraviolet (EUV) reflectivity of the samples with 20 bilayers gradually decreased from 26.3% to 18.9% with increased working pressure. The bandwidth of the reflection peak remains similar for the different samples with a full width half-maximum (FWHM) value of around 0.87 nm. A maximum EUV reflectivity of 33.2% and a bandwidth of 0.64 nm were achieved by the sample with 50 bilayers fabricated under a working pressure of 0.13 Pa.

A free-standing thin foil bolometer for measuring soft x-ray fluence.

A free-standing thin foil bolometer for measuring soft x-ray fluence in z-pinch experiments is developed. For the first time, we present the determination of its sensitivity by different methods. The results showed great consistency for the different methods, which confirms the validity of the sensitivity and provides confidence for its application in z-pinch experiments. It should be highlighted that the sensitivity of a free-standing foil bolometer could be calibrated directly using Joule heating without any corrections that will be necessary for a foil bolometer with substrate because of heat loss. The difference of the waveforms between the free-standing foil bolometer and that with substrate is obvious. It reveals that the heat loss to the substrate should be considered for the latter in despite of the short x-ray pulse when the peak value is used to deduce the total deposited energy. The quantitative influence is analyzed through a detailed simulation.

Measurements of high energy photons in Z-pinch experiments on primary test stand.

High energy photons are measured for the first time in wire-array Z-pinch experiments on the Primary Test Stand (PTS) which delivers a current up to 8 MA with a rise time of 70 ns. A special designed detecting system composed of three types of detectors is used to measure the average energy, intensity, and pulse waveform of high energy photons. Results from Pb-TLD (thermoluminescence dosimeter) detector indicate that the average energy is 480 keV (±15%). Pulse shape of high energy photons is measured by the photodiode detector consisted of scintillator coupled with a photodiode, and it is correlated with soft x-ray power by the same timing signal. Intensity is measured by both TLD and the photodiode detector, showing good accordance with each other, and it is 10(10) cm(-2) (±20%) at 2 m in the horizontal direction. Measurement results show that high energy photons are mainly produced in pinch regions due to accelerated electrons. PTS itself also produces high energy photons due to power flow electrons, which is one order smaller in amplitude than those from pinch region.

Elevated levels of mitochonrial respiratory complexes activities and ATP production in 17-ß-estradiol-induced prolactin-secretory tumor cells in male rats are inhibited by melatonin in vivo and in vitro.

BACKGROUND: Our earlier studies indicate that melatonin inhibits the proliferation of prolactinoma and induces apoptosis of pituitary prolactin-secreting tumor in rats. Melatonin has also been shown to induce apoptosis and to reduce the production of ATP in breast tumor cells. This study analyzed the levels of the four mitochondrial respiratory complexes and the production of ATP and also the effects of melatonin treatment of prolactinoma. METHODS: In the in vivo study, mitochondria were harvested from control pituitaries or prolactinoma collected from the pituitaries of melatonin- and 17-ß-estradiol (E2)-treated male rats. In the in vitro study, prolactinoma cells mitochondria were harvested. Activities of the four mitochondrial respiratory complexes were assayed using fluorometer. ATP production of prolactinoma cells was estimated using bioluminescent methods. RESULTS: Elevated levels of four mitochondrial respiratory complexes activities and ATP production were recorded in prolactinoma cells. Moreover, in both in vivo and in vitro studies, melatonin inhibited the activities of mitochondrial respiratory complexes and the production of ATP in prolactinoma cells. CONCLUSIONS: There is a link between mitochondrial function increase and tumorigenesis. Melatonin induces apoptosis of pituitary prolactin-secreting tumor of rats via the induction of mitochondrial dysfunction and inhibition of energy metabolism.

A novel extreme ultraviolet four channels normal incidence imaging system for plasma diagnostics of Z-pinch facility.

A novel EUV four channels normal incidence imaging system for plasma diagnostics of Z-pinch facility was presented in this paper, which consists of four concave mirrors and one convex mirror used for focusing an object onto four different positions with about 30 µm resolution on the same image plane. In addition, this imaging system can work at the energies of 50 eV, 95 eV, 150 eV, and broadband of 50-100 eV by using different multilayer films deposited on the concave and convex mirrors. This instrument, combined with framing camera, can achieve the power of two-dimensional spatial and temporal resolution, as well as the ability to imaging the plasma at the specific temperature. In the paper, the four channels microscope centering at multi-energies was developed.

Chronic administration of hexarelin attenuates cardiac fibrosis in the spontaneously hypertensive rat.

Cardiac fibrosis is a hallmark of heart disease and plays a vital role in cardiac remodeling during heart diseases, including hypertensive heart disease. Hexarelin is one of a series of synthetic growth hormone secretagogues (GHSs) possessing a variety of cardiovascular effects via action on GHS receptors (GHS-Rs). However, the role of hexarelin in cardiac fibrosis in vivo has not yet been investigated. In the present study, spontaneously hypertensive rats (SHRs) were treated with hexarelin alone or in combination with a GHS-R antagonist for 5 wk from an age of 16 wk. Hexarelin treatment significantly reduced cardiac fibrosis in SHRs by decreasing interstitial and perivascular myocardial collagen deposition and myocardial hydroxyproline content and reducing mRNA and protein expression of collagen I and III in SHR hearts. Hexarelin treatment also increased matrix metalloproteinase (MMP)-2 and MMP-9 activities and decreased myocardial mRNA expression of tissue inhibitor of metalloproteinase (TIMP)-1 in SHRs. In addition, hexarelin treatment significantly attenuated left ventricular (LV) hypertrophy, LV diastolic dysfunction, and high blood pressure in SHRs. The effect of hexarelin on cardiac fibrosis, blood pressure, and cardiac function was mediated by its receptor, GHS-R, since a selective GHS-R antagonist abolished these effects and expression of GHS-Rs was upregulated by hexarelin treatment. In summary, our data demonstrate that hexarelin reduces cardiac fibrosis in SHRs, perhaps by decreasing collagen synthesis and accelerating collagen degradation via regulation of MMPs/TIMP. Hexarelin-reduced systolic blood pressure may also contribute to this reduced cardiac fibrosis in SHRs. The present findings provided novel insights and underscore the therapeutic potential of hexarelin as an antifibrotic agent for the treatment of cardiac fibrosis.

Hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in the rat.

Growth hormone-releasing peptides (GHRP) and ghrelin are synthetic and natural ligands of growth hormone secretagogue receptor (GHSR) respectively and are shown to exert protective actions on cardiac dysfunction. Because ghrelin has been reported to inhibit proinflammatory responses in human endothelium and GHSR has been identified in blood vessels, we hypothesized that GHRP could alleviate the development of atherosclerosis (As). Atherosclearosis was induced by a short period (4 days) of vitamin D(3) and chronic (three months) intragastric feeding of high fat emulsion (containing 0.5% propylthiouracil) in adult SD rats. Some As rats received chronic hexarelin (a variant of GHRP) injection (SC BID, 30 days) and normal rats received placebo as control. Significant atherosclerosis developed in animals fed with the emulsion. Serum total cholesterol and LDL-c increased, and HDL-c and aortic nitric oxide (NO) decreased significantly in As group. Hexarelin suppressed the formation of atherosclerotic plaques and neointima, partially reversed serum HDL-c/LDL-c ratio and increased the levels of serum NO and aortic mRNAs of eNOS, GHSR and CD36 in As rats. Hexarelin also decreased [(3)H]-TdR incorporation in cultured vascular smooth muscle cell (VSMC) and calcium sedimentation in aortic wall. Furthermore, foam cell formation induced by ox-LDL was decreased by hexarelin. In conclusion, hexarelin suppresses high lipid diet and vitamin D3-induced atherosclerosis in rats, possibly through upregulating HDL-c/LDL-c ratio, vascular NO production and downregulating the VSMC proliferation, aortic calcium sedimentation and foam cell formation. These novel anti-atherosclerotic actions of hexarelin suggest that the peptide might have a clinical potential in treating atherosclerosis.

A time-resolved spectroscopic diagnostic based on fast scintillator and optical fiber array for z-pinch plasmas.

We report a specially designed type of temporal resolved x-ray spectroscopic diagnostic using a spherically bent quartz crystal for z-pinch plasmas. Registration of time-resolved spectra was accomplished by coupling fast plastic scintillator, an optical fiber array, an optical streak camera, and a charge coupled device as the recording medium of this diagnostic. The diagnostic has been tested in imploding wire array experiments on S-300 pulsed power facility. Time-resolved K-shell lines were successfully obtained for aluminum wire array implosion plasmas.

Antiproliferative effects of melatonin on the growth of rat pituitary prolactin-secreting tumor cells in vitro.

Earlier studies showed that melatonin reduced the growth of 17-beta-estradiol (E(2))-induced rat pituitary prolactin-secreting tumor (prolactinoma) in vivo. The mechanisms of melatonin's inhibitory action on the prolactin-secreting tumor were further explored by investigating the in vitro effects of melatonin on the growth of pituitary prolactin-secreting tumor cells. Primary cultured prolactinoma cells from E(2)-induced rat pituitary prolactin-secreting tumor were treated with 10(-5), 10(-4) or 10(-3) m melatonin for 5 days. Apoptosis was evaluated using flow cytometry and the TdT-mediated dUTP nick-end labeling (TUNEL) method. In addition, cell viability was analyzed by (3,4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. It was found that incubation of prolactinoma cells with 10(-5), 10(-4) or 10(-3) m melatonin for 5 days inhibited cell growth and increased cell apoptosis. Furthermore, melatonin increased caspase-3 activity, Bax mRNA expression, and cytochrome c protein expression. Conversely, Bcl-2 mRNA expression and mitochondrial membrane potential were inhibited by melatonin treatment. Our results further suggest that melatonin inhibits tumor growth by inducing apoptosis of rat pituitary prolactin-secreting tumor directly via the damage of mitochondria.

Inhibitory effects of melatonin on the growth of pituitary prolactin-secreting tumor in rats.

The in vivo effects of melatonin on proliferation and apoptosis of 17-beta-estradiol (E2)-induced pituitary prolactin-secreting tumor (prolactinoma) were investigated in rats kept in 12 L/12 D (lights on: 06:00-18:00 hr). As melatonin was shown to induce apoptosis of breast and liver tumor cells, we examined whether melatonin would induce apoptosis of rat pituitary prolactinoma cells. 0.125, 0.25, 0.50 or 1.0 mg melatonin/day/rat was administrated subcutaneously at 17:30-18:00 hr. The weight of prolactinomas was measured. Apoptosis was evaluated using the TdT-mediated dUTP nick-end labeling method. It was found that treatment with 0.25 and 0.50 mg melatonin for 97 days inhibited prolactinoma cell proliferation and increased prolactinoma cell apoptosis. Furthermore, melatonin induced mRNA expression of Bax and cytochrome c protein expression. Conversely, mRNA expression of Bcl-2, and mitochondrial membrane potential were inhibited by melatonin treatment. These results suggest that melatonin inhibits the proliferation and induces apoptosis of rat pituitary prolactin-secreting tumor via perturbation of mitochondria physiology.

[Melatonin suppressing the proliferation of E2-induced pituitary prolactin-secreting tumor in rat involves the effects of estrogen receptor].

AIM: To investigate effects of melatonin on estrogen receptor at the primary stage of melatonin (MLT) inhibiting the proliferation of 17-beta-estradiol (E2)-induced pituitary prolactin-secreting tumor (prolactinoma) and its mechanisms in the rat. METHODS: MLT inhibiting the proliferation of 17-beta-E2-induced prolactinoma was created by administrating different concentration of melatonin subcutaneously at 18:00 in every group of SD rat in vivo. We also examined the expression of MLT receptor in prolactinoma cells and the effects of MLT on the expression of estrogen receptor (ER) by in situ hybridization and the effects of MLT on the binding of ER to estrogen response element (ERE) by electrophoretic mobility shift assay (EMSA)in primary culture cells iv vitro. RESULTS: The results showed that the weights of prolactinomas in MLT groups, in which 0.25 mg or 0.50 mg/day/rat melatonin was administrated subcutaneously at 18:00, were decreased significantly (P < 0.01 and P < 0.05). The expression of MLT1a and MLT1b were shown in pituitary prolactinoma cells. Compared with the prolactinoma, the expression of ER and the bind of ER to ERE in prolactinoma treated with 0.25 mg/day/rat or 0.50 mg/day/rat MLT was decreased (P < 0.01 and P < 0.01). CONCLUSION: These data indicate that some dosage of MLT inhibit the development of E2-induced prolactinoma in SD rat. One of the mechanisms is involved in suppressing the expression of estrogen receptor and partly inhibiting the bind of ER to ERE.

[Analysis of different mutations in regulatory sequence of prolactin gene during the formation of 17 beta-estradiol-induced prolactinoma in eutopic and ectopic pituitary of rats].

OBJECTIVE: To analyze different mutations in regulatory sequence of prolactin (PRL) gene during the formation of 17 beta-estradiol (E2 ) -induced prolactinoma in eutopic and ectopic pituitary of rats. METHODS: Male Sprague-Dawley rats transplanted with an isologaus pituitary under renal capsule were treated with subcutaneous implantation of an empty or E2-laden silastic capsule. Reverse transcription-polymerase chain reaction was employed to evaluate the expression of PRL mRNA in pituitary glands, and DNA sequencing was used to analyze the mutation in regulatory sequence of PRL gene. RESULTS: After treated with E2 for 120 days, both the eutopic and ectopic pituitaries were three times more heavier than those from control group (P < 0. 01) , and the body weight of rats was decreased to 42. 90% of the control group (P < 0 01 ). The PRL mRNA expressions in glands from the eutopic and ectopic pituitaries 120 days after treated with E2 were much more than those in untreated pituitary glands (P <0. 01). DNA sequencing showed seven mutations in the regulatory sequence of PRL gene in the eutopic pituitaries 120 days after treated with E2 , while the mutation in the ectopic pituitaries was decreased. CONCLUSIONS: Prolactinomas can be induced by chronic treatment with E2 in both the eutopic and the ectopic pituitaries transplanted under renal capsule distant from the hypothalamus. Different mechanisms exist in the formation of eutopic and ectopic prolactinomas.

[Melatonin inhibits the proliferation of pituitary prolactin-secreting tumor by suppressing the enhancer elements mutation of PRL gene in the rat].

In order to investigate the molecular mechanisms of the inhibition of the proliferation of 17-beta-estradiol (E(2))-induced pituitary prolactin-secreting tumor (prolactinoma) by melatonin (MLT) in the rat, we examined the inhibitory effects of MLT on the proliferation of E(2)-induced prolactinoma of the rat and the suppressing effects of MLT on the enhancer elements mutation of PRL gene in vivo and in vitro. The results showed that the weights of prolactinomas in MLT groups, in which 0.25 mg or 0.50 mg per day per rat of MLT was administered subcutaneously at 18:00, were decreased significantly. Out of the dosage of MLT, such as 0.05, 1.00 mg and 2.00 mg per day per rat, the antitumor action of MLT is less or disappointing. Polymerase chain reaction (PCR) and DNA sequencing showed five mutations in the enhancer elements of PRL gene in prolactinoma, such as -1885 point mutation (C --> G), -1857 - -1855 substitution (ACA --> G), -1792 - -1791 insertion G, -1383 - -1382 insertion (GGTGTGTG), -1265 - -1250 deletion (GTGTGTGTGTGTGTGT). Excluding of -1885 point mutation (C --> G), the mutation in the prolactinoma treated with 0.25 mg per day per rat MLT was decreased, such as -1792 - -1791 without insertion of G, -1856 - -1855 deletion AC, -1385 - -1384 deletion TG, -1250 - -1253 deletion GTGT. Firefly luciferase reporter gene systems showed that the luminosity of enhancer elements-luciferase reporter fusion gene in normal pituitary, prolactinoma treated without or with 0.25 mg per day per rat MLT were (13448.17+/-3012.74), (161831.67+/-60996.01), and (10212.17+/-634.71) OD units. Compared with the normal pituitary, the activity of PRL gene enhancer elements in prolactinoma was increased by 11 times (P<0.001). Compared with the prolactinoma, the activity of PRL gene enhancer elements in prolactinoma treated with MLT was decreased by 93.69% (P<0.001). Analysis of the space structure of PRL gene enhancer elements showed that the bending index in prolactinoma was higher than that in prolactinoma treated with MLT, which was higher than that in the normal pituitary. These results demonstrate that one of the important molecular mechanisms of MLT inhibiting the proliferation of prolactinoma is related to the reduction of enhancer elements mutation of PRL gene. These data also suggest that MLT-induced attenuation of enhancer elements mutation of PRL gene is involved in decreasing the bending index and attenuating the higher expression of PRL gene.