Changes in gap junction proteins Connexin30. 2 and Connexin40 expression in the sinoatrial node of rats with dexmedetomidine-induced sinus bradycardia

Abstract Background Dexmedetomidine (Dex) is widely used, and its most common side effect is bradycardia. The complete mechanism through which Dex induces bradycardia has not been elucidated. This research investigates the expression of gap junction proteins Connexin30.2 (Cx30.2) and Connexin40 (Cx40) within the sinoatrial node of rats with Dex-induced sinus bradycardia. Methods Eighty rats were randomly assigned to five groups. Saline was administered to rats in Group C. In the other four groups, the rats were administered Dex to induce bradycardia. In groups D1and D2, the rats were administered Dex at a loading dose of 30 μg.kg−1 and 100 μg.kg−1 for 10 min, then at 15 μg.kg−1.h−1 and 50 μg.kg−1.h−1 for 120 min separately. The rats in group D1A and D2A were administered Dex in the same way as in group D1and D2; however, immediately after the administration of the loading dose, 0.5 mg atropine was administered intravenously, and then at 0.5 mg.kg−1.h−1 for 120 min. The sinoatrial node was acquired after intravenous infusion was completed. Quantitative real-time polymerase chain reaction and western blot analyses were performed to measure mRNA and protein expression of Cx30.2 and Cx40, respectively. Results The expression of Cx30.2 increased, whereas the expression of Cx40 decreased within the sinoatrial node of rats with Dex-induced sinus bradycardia. Atropine reversed the effects of Dex on the expression of gap junction proteins. Conclusion Dex possibly altered the expression of gap junction proteins to slow down cardiac conduction velocity in the sinoatrial node.

An Arg124Cys mutation in transforming growth factor ?-induced gene associated with lattice corneal dystrophy type I in a Chinese pedigree

Purpose: To identify a clinical and genetic form of a large Chinese family with an autosomal?dominant lattice corneal dystrophy type I (LCD I). Methods: The patients’ eyes were examined on the basis of slit?lamp microscopy, and other clinical records were also collected. Genomic DNA was extracted from peripheral leukocytes of the affected patients and their unaffected family members. Each previous reported mutation of the transforming growth factor ??induced gene (TGFBI) gene was amplified by touch?down polymerase chain reaction and directly sequenced to verify the disease?causing mutation. Results: Typical clinical features of LCD I were found by slit?lamp photography in these affected Chinese pedigrees. A heterozygous single base?pair transition from C to T (c.418 C > T), leading to amino acid substitution Arg124Cys (R124C) in the encoded TGFBI protein, was detected in all of the eighteen affected patients. The same mutation was not found in unaffected family members. Conclusion: The R124C mutation hot spot, which was relatively rare in China, was responsible for LCD I in the large family. Molecular genetic analysis of TGFBI gene can offer an accurate diagnosis of patients with lattice corneal dystrophies in the clinical treatment.

Value of metagenomic next-generation sequencing in children with severe infectious diseases / 中国当代儿科杂志

OBJECTIVES@#To study the application value of metagenomic next-generation sequencing (mNGS) in children with severe infectious diseases.@*METHODS@#An analysis was performed on the clinical data and laboratory test results of 29 children with severe infection who were admitted to the Second Affiliated Hospital of Wenzhou Medical University from June 2018 to December 2020. Conventional pathogen culture was performed for the 29 specimens (27 peripheral blood specimens and 2 pleural effusion specimens) from the 29 children, and mNGS pathogen detection was performed at the same time.@*RESULTS@#Among the 29 children, 2 tested positive by conventional pathogen culture with 2 strains of pathogen, and the detection rate was 7% (2/29); however, 20 children tested positive by mNGS with 38 strains of pathogen, and the detection rate was 69% (20/29). The pathogen detection rate of mNGS was significantly higher than that of conventional pathogen culture (P<0.05), and mNGS could detect the viruses, fungi, and other special pathogens that conventional pathogen culture failed to detect, such as Orientia tsutsugamushi. The univariate analysis showed that gender, routine blood test results, C-reactive protein, procalcitonin, D-dimer, radiological findings, and whether antibiotics were used before admission did not affect the results of mNGS (P>0.05).@*CONCLUSIONS@#Compared with conventional pathogen culture, mNGS is more sensitive for pathogen detection, with fewer interference factors. Therefore, it is a better pathogenic diagnosis method for severe infectious diseases in children.

Impact of neoadjuvant chemoradiotherapy on the local recurrence and distant metastasis pattern of locally advanced rectal cancer: a propensity score-matched analysis / 中华医学杂志(英文版)

BACKGROUND@#Previous studies have demonstrated different predominant sites of distant metastasis between patients with and without neoadjuvant chemoradiotherapy (NCRT). This study aimed to explore whether NCRT could influence the metastasis pattern of rectal cancer through a propensity score-matched analysis.@*METHODS@#In total, 1296 patients with NCRT or post-operative chemoradiotherapy (PCRT) were enrolled in this study between January 2008 and December 2015. Propensity score matching was used to correct for differences in baseline characteristics between the two groups. After propensity score matching, the metastasis pattern, including metastasis sites and timing, was compared and analyzed.@*RESULTS@#After propensity score matching, there were 408 patients in the PCRT group and 245 patients in the NCRT group. NCRT significantly reduced local recurrence (4.1% vs. 10.3%, P = 0.004), but not distant metastases (28.2% vs. 27.9%, P = 0.924) compared with PCRT. In both the NCRT and PCRT groups, the most common metastasis site was the lung, followed by the liver. The NCRT group developed local recurrence and distant metastases later than the PCRT group (median time: 29.2 [18.8, 52.0] months vs. 18.7 [13.3, 30.0] months, Z = -2.342, P = 0.019; and 21.2 [12.2, 33.8] vs. 16.4 [9.3, 27.9] months, Z = -1.765, P = 0.035, respectively). The distant metastases occurred mainly in the 2nd year after surgery in both the PCRT group (39/114, 34.2%) and NCRT group (21/69, 30.4%). However, 20.3% (14/69) of the distant metastases appeared in the 3rd year in the NCRT group, while this number was only 13.2% (15/114) in the PCRT group.@*CONCLUSIONS@#The predominant site of distant metastases was the lung, followed by the liver, for both the NCRT group and PCRT group. NCRT did not influence the predominant site of distant metastases, but the NCRT group developed local recurrence and distant metastases later than the PCRT group. The follow-up strategy for patients with NCRT should be adjusted and a longer intensive follow-up is needed.

Advances of Lefamulin: a new pleuromutilin antibiotic / 药学学报

Lefamulin (BC-3781) is a semi-synthetic pleuromutilin antibiotic, approved for the treatment of community-acquired bacterial pneumonia (CABP) by Food and Drug Administration (USA) in August 2019, with the commodity name of Xenleta. It is the first pleuromutilin antibiotics used for systemic treatment of bacterial infections in human. Lefamulin binds to the peptidyl transferase center of the 50S ribosomal subunit to prevent peptide transfer, thus inhibits protein synthesis. Lefamulin displays expanded activity against gram-positive organisms, and also shows high activity against atypical microorganism like Mycoplasma pneumoniae. This review discusses the mechanism, bacterial spectrum of activity, preclinical and clinical data of Lefamulin.

Safety analysis of definitive surgery for chronic radiation intestinal injury / 中华胃肠外科杂志

Objective: To investigate the safety of definitive surgery for chronic radiation intestinal injury. Methods: A descriptive case series study was performed. Clinical data of 105 patients who were diagnosed as chronic radiation intestinal injury, had complete data and received definitive surgery (the radiation-induced intestinal segment and digestive tract reconstruction) at Department of Gastrointestinal Surgery of Beijing Tsinghua Changgung Hospital from June 2016 to May 2020 were retrospectively analyzed. There were 30 males (28.6%) and 75 females (71.4%) with the median age of 58 years (P25, P75: 52, 64 years). Patients who had tumor recurrence or refused surgical treatment were excluded. According to the preoperative evaluation and clinical manifestations, to select the resection range. Outcome parameters: (1) preoperative evaluation (nutrition risk assessment and status of obstruction or fistula); (2) clinical manifestations and treatment strategies; (3) details of surgical parameters; (4) postoperative complications, and Clavien-Dindo classification III to V was defined as main moderate-severe complication. Results: (1) Preoperative evaluation: Eighty-eight patients (83.8%) developed symptoms of chronic radiation intestinal injury more than 1 year after the end of radiotherapy. Ninety-eight patients (93.3%) had preoperative NRS-2002 score ≥3, 74 patients (70.5%) received preoperative parenteral nutritional support, and the median time of nutritional support was 10.5 (7.0, 16.0) days. Sixteen patients (15.2%) received small intestinal decompression tube implantation due to severe obstruction. (2) Clinical manifestations and treatment strategies: Among 105 patients, 87 (82.9%) presented with obstruction and received definitive resection of the radiation-induced intestinal segment plus one-stage digestive tract reconstruction; 18 (17.1%) presented with intestinal fistula and all of them received definitive resection of the radiation-induced intestinal segment, intestinal fistula plus one-stage digestive tract reconstruction. Among above 18 patients with fistula, 3 patients with ileorectal stump fistula received pedicled pelvic closure of greater omentum at the same time; 4 patients had ileal vesical fistula, of whom 2 patients received cystectomy and bladder repair due to preoperative nephrostomy decompression, and the other 2 patients received transection of the small intestine proximal and distal to the fistula and anastomosis of the intestinal loop without fistula resection, intestinal fistula or bladder fistula repair. (3) The details of surgical parameters: Median operative time and intraoperative blood loss was 230 (180, 300) minutes and 50 (20, 50) ml respectively. Ninety-two patients (92/105, 87.6%) underwent ileocolonic anastomosis, and anastomosis on the hepatic flexure or splenic flexure colon were performed in 88 (83.8%) and 4 (3.8%) patients respectively. Ileoileal anastomosis was performed in 13 patients (12.4%). The anastomotic site of 92 patients (87.6%) was strictly located in the contralateral quadrant of the radiation field, and the anastomotic site of 13 patients (12.4%) was far from the radiation field. Nine patients (8.6%) had more than one anastomosis, 5 patients (4.8%) had less than 180 cm of residual small intestine, 7 patients (6.7%) underwent retrograde intestinal permutation, 4 patients (3.8%) underwent abdominal wall reconstruction surgery due to abdominal wall defects, and 87 patients (82.9%) had severe abdominal pelvic adhesions (grade 3-4 adhesions). Intraoperative complications occurred in 3 patients (2.9%), which were found in time and handled properly. The median postoperative hospital stay was 13.0 (12.0, 24.5) days, and all the patients had resumed oral feeding upon discharge. (4) Postoperative complications: Fourteen patients (13.3%) had 18 major complications (grade III to V). The incidence of postoperative anastomotic leakage was 5.7% (6/105), and the incidence of anastomotic leakage for ileocolon anastomosis and ileoileal anastomosis was 2.2% (2/92) and 4/13, respectively (χ(2)=17.29, P<0.001). The incidence of postoperative anastomotic leakage of intestinal fistula and intestinal obstruction was 3/18 and 3.4% (3/87), respectively (χ(2)=4.84, P=0.028). The mortality at 30 days after operation was 1.0% (1/105), after abdominal infection and septic shock caused by postoperative anastomotic leakage resulting in multiple organ failure. Conclusion: For chronic radiation intestinal injury patients with obstruction or fistula, definitive surgical treatment is feasible and safe with acceptable major complications.

Advances in research of application of low-temperature plasma in microbiology / 上海预防医学

Low-temperature plasma has special physicochemical properties that can cause inactivation and mutation of microorganisms through complex biochemical processes.This article reviews the application of low-temperature plasma in microbiology, including sterilization, screening of bacterial strains, and antibacterial treatment.It also discussed the current research status and existing problems to provide ideas for promoting the development of interdisciplinary collaboration in low-temperature plasma reserch.

New cases are they incident or newly discovered / 中华疾病控制杂志

@#New case is a basic concept in epidemiology and often serves as the numerator for the indexes of the frequency of disease in a population． However，the exact definition of new case is still un- clear． Based on the natural history of disease combined with the concept of event-state，this article rede- fines case-related concepts and compares them with traditional concepts． The study found that the so- called new cases in traditional epidemiological studies are actually the cases firstly discovered，including newly discovered cases and identifiable newly incident cases ( disease initiation) ． In the real world，new cases are often difficult to measure，and together with the continuous process of disease development and iceberg phenomenon，those may affect the accuracy of disease frequency measurement and causal infer- ence． A correct understanding of the meaning of case-related concepts，the meanings they reflect，and the differences between them help to rationally interpret the results of epidemiological research．

UPLC-Q-TOF-MS Analysis of Chemical Constituents of Paederiae Herba for Folk Use / 中国实验方剂学杂志

Objective:The chemical constituents of ethanol extracts from Paederia scandens，P. scandens var.tomentosa，P. stenophylla and P. foetida used in folk were identified. And the differences of chemical constituents among the four kinds of ethanol extracts were compared. Method:The chemical constituents of four species of Paederia were qualitatively and rapidly analyzed by UPLC-Q-TOF-MS. The data were analyzed manually and matched through databases to determine the differences among the chemical constituents of four species of Paederia. Result:Twenty-three compounds were identified from four species, including 12 iridoid glycosides，such as paederoside，paederosidic acid，asperuloside，asperulosidic acid，paederosidic acid methyl ester and monotropein.5 quinic acid derivatives, such as 1-O-caffeoylquinic acid，chlorogenic acid，cryptochlorogenic acid,and 5 flavonoid glycosides，such as quercetin-3-O-rutinoside-7-O-glucoside,quercetin-3-O-rutinoside-7-O-xyloside，rutin and simple compoundkinsenoside. Conclusion:Based on the UPLC-Q-TOF-MS analysis of the chemical components of four kinds of Paederia，the four peaks with the highest response intensity were paederoside，paederosidic acid，asperuloside and asperulosidic acid，all of which were iridoid glycosides.

Application of I-gel laryngeal mask airway in EBUS-TBNA performed under intravenous anesthesia / 中国内镜杂志

Objective To seek better anesthesia management methods for clinical applications guidance by retrospectively analyze the clinical effects of I-gel laryngeal mask for endobronchial ultrasound-guided transbronchial needleaspiration (EBUS-TBNA) performed under local and intravenous anesthesia modes. Methods Review the differences in the operative time, the number of SpO2 <95%, body motion, cough, laryngospasm, nausea, surgical puncture, operator suspension, postoperative throat discomfort, and satisfaction scores among the patient, the operator and the anesthetist between the two kinds of anesthesia of 60 patients for EBUS-TBNA in 3 years from 2013 to 2016. Results From June 2014, patients with I-gel laryngeal mask for EBUS-TBNA under intravenous anesthesia had fewer cough, nausea, physical activity, shorter operative time, and higher anesthesia satisfaction scores. Conclusion The perioperative management method of I-gel laryngeal mask for EBUS-TBNA in intravenous anesthesia is more secure and more feasible, recommended for clinical use.

Account for Clinical Heterogeneity in Assessment of Catheter-based Renal Denervation among Resistant Hypertension Patients: Subgroup Meta-analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Catheter-based renal denervation (RDN) is a novel treatment for resistant hypertension (RH). A recent meta-analysis reported that RDN did not significantly reduce blood pressure (BP) based on the pooled effects with mild to severe heterogeneity. The aim of the present study was to identify and reduce clinical sources of heterogeneity and reassess the safety and efficacy of RDN within the identified homogeneous subpopulations.</p><p><b>METHODS</b>This was a meta-analysis of 9 randomized clinical trials (RCTs) among patients with RH up to June 2016. Sensitivity analyses and subgroup analyses were extensively conducted by baseline systolic blood pressure (SBP) level, antihypertensive medication change rates, and coronary heart disease (CHD).</p><p><b>RESULTS</b>In all patients with RH, no statistical differences were found in mortality, severe cardiovascular events rate, and changes in 24-h SBP and office SBP at 6 and 12 months. However, subgroup analyses showed significant differences between the RDN and control groups. In the subpopulations with baseline 24-h SBP ≥155 mmHg (1 mmHg = 0.133 kPa) and the infrequently changed medication, the use of RDN resulted in a significant reduction in 24-h SBP level at 6 months (P = 0.100 and P= 0.009, respectively). Subgrouping RCTs with a higher prevalent CHD in control showed that the control treatment was significantly better than RDN in office SBP reduction at 6 months (P < 0.001).</p><p><b>CONCLUSIONS</b>In all patients with RH, the catheter-based RDN is not more effective in lowering ambulatory or office BP than an optimized antihypertensive drug treatment at 6 and 12 months. However, among RH patients with higher baseline SBP, RDN might be more effective in reducing SBP.</p>

Resolvin D1 Protects Lipopolysaccharide-induced Acute Kidney Injury by Down-regulating Nuclear Factor-kappa B Signal and Inhibiting Apoptosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Resolvin D1 (RvD1) is a newly found anti-inflammatory bioactive compound derived from polyunsaturated fatty acids. The current study aimed to explore the protective effect of RvD1 on lipopolysaccharide (LPS)-induced acute kidney injury (AKI) and its possible mechanism.</p><p><b>METHODS</b>Both in vivo and in vitro studies were conducted. Male BALB/c mice were randomly divided into control group (saline), LPS group (LPS 5 mg/kg), RvD1 group (RvD1 5 μg/kg + LPS 5 mg/kg), and blockage group (Boc-MLP 5 μg/kg + RvD1 5 μg/kg + LPS 5 mg/kg). Boc-MLP is a RvD1 receptor blocker. The mice were intraperitoneally injected with these drugs and recorded for general condition for 48 h, while the blood and kidneys were harvested at 2, 6, 12, 24, and 48 h time points, respectively (n = 6 in each group at each time point). Human proximal tubule epithelial cells (HK-2) were randomly divided into control group (medium only), LPS group (LPS 5 μg/ml), RvD1 group (RvD1 10 ng/ml + LPS 5 μg/ml), and blockage group (Boc-MLP 10 ng/ml + RvD1 10 ng/ml + LPS 5 μg/ml). The cells were harvested for RNA at 2, 4, 6, 12, and 24 h time points, respectively (n = 6 in each group at each time point). Blood creatinine was tested by using an Abbott i-STAT portable blood gas analyzer. Tumor necrosis factor-α (TNF-α) level was detected by ELISA. Kidney pathology was observed under hematoxylin and eosin (HE) staining and transmission electron microscope (TEM). We hired immune-histological staining, Western blotting, and fluorescence quantitative polymerase chain reaction to detect the expression of RvD1 receptor ALX, nuclear factor-kappa B (NF-κB) signaling pathway as well as caspase-3. Kidney apoptosis was evaluated by TUNEL staining.</p><p><b>RESULTS</b>RvD1 receptor ALX was detected on renal tubular epithelials. Kaplan-Meier analysis indicated that RvD1 improved 48 h animal survival (80%) compared with LPS group (40%) and RvD1 blockage group (60%), while RvD1 also ameliorated kidney pathological injury in HE staining and TEM scan. After LPS stimulation, the mRNA expression of toll-like receptor 4, myeloid differentiation factor 88, and TNF-α in both mice kidneys and HK-2 cells were all up-regulated, while RvD1 substantially inhibited the up-regulation of these genes. Western blotting showed that the phosphorylated-IκB/IκB ratio in LPS group was significantly higher than that in the control group, which was inhibited in the RvD1 group. RvD1 could inhibit the up-regulation of cleaved-caspase-3 protein stimulated by LPS, which was prohibited in RvD1 blockage group. RvD1 group also had a lower proportion of apoptotic nuclei in mice kidney by TUNEL staining compared with LPS group.</p><p><b>CONCLUSION</b>In LPS-induced AKI, RvD1 could decrease TNF-α level, ameliorate kidney pathological injury, protect kidney function, and improve animal survival by down-regulating NF-κB inflammatory signal as well as inhibiting renal cell apoptosis.</p>

The first report of the vanC1 gene in Enterococcus faecium isolated from a human clinical specimen

The vanC1 gene, which is chromosomally located, confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Enterococcus faecium TJ4031 was isolated from a blood culture and harbours the vanC1gene. Polymerase chain reaction (PCR) assays were performed to detect vanXYc and vanTc genes. Only the vanXYc gene was found in the E. faecium TJ4031 isolate. The minimum inhibitory concentrations of vancomycin and teicoplanin were 2 µg/mL and 1 µg/mL, respectively. Real-time reverse transcription-PCR results revealed that the vanC1and vanXYc genes were not expressed. Pulsed-field gel electrophoresis and southern hybridisation results showed that the vanC1 gene was encoded in the chromosome. E. faecalis isolated from animals has been reported to harbour vanC1gene. However, this study is the first to report the presence of the vanC1gene in E. faecium of human origin. Additionally, our research showed the vanC1gene cannot serve as a species-specific gene of E. gallinarum and that it is able to be transferred between bacteria. Although the resistance marker is not expressed in the strain, our results showed that E. faecium could acquire the vanC1gene from different species.

Post-stroke shoulder-hand syndrome treated with acupuncture and rehabilitation: a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To assess the clinical efficacy on post-stroke shoulder-hand syndrome (SHS) treated with acupuncture and rehabilitation and the impacts on patients' nailfold microcirculation.</p><p><b>METHODS</b>One hundred and twenty patients were randomized into an acupuncture rehabilitation group and a simple rehabilitation group, 60 cases in each one. In the simple rehabilitation group, OT (comprehensive rehabilitation therapy) training was adopted. In the acupuncture rehabilitation group, on the basis of the treatment as the simple rehabilitation, acu puncture was added at Taiyuan (LU 9), Zusanli (ST 36), Xuanzhong (GB 39), Waiguan (TE 5), Shousanli (LI 10), Quchi (LI 11) and Jianyu (LI 15). Acupuncture was given once a day, 7 days made one session. Totally, 4 sessions of treatment were required. Fugl-Meyer score, upper limb pain score, the score of nerve function defect and the items of nailfold microcirculation of patients were assessed in the the two groups before and after treatment. The efficacy was compared between the two groups.</p><p><b>RESULTS</b>(1) The upper limb pain, the systematic motor function of the upper limbs, the nerve function defect, nailfold microcirculation and clinical symptoms were all improved after treatment in either the acupuncture rehabilitation group or the simple rehabilitation group as compared with those before treatment, indicating the significant difference (P<0. 05, P<0. 01). (2) The im provements in the upper limb pain (0. 90+/-0.71 vs 1. 80+/-0. 66), the systematic motor function of the upper limbs (42. 43 13. 57 vs 29. 98+/-15. 11), the nerve function defect (8. 60+/-11. 61 vs 13. 0+/-1. 74), nailfold microcirculation (total score 3. 18+/-1.32 vs 4.34+/-1.23) and clinical symptoms in the acupuncture rehabilitation group after treatment were different significantly as compared with those in the simple rehabilitation group (PO0. 05,P-O. 01), and the results in the acupuncture rehabilitation group were superior to the simple rehabilitation group. (3) In the acupuncture rehabilitation group, the markedly effective rate was 50. 0% (30/60) and the total effective rate was 93.3% (56/60), which was better than 16.7% (10/60) and 63. 3% (38/60) respectively in the simple rehabilitation group (all P<0. 05).</p><p><b>CONCLUSION</b>Both the combined therapy of acupuncture and rehabilitation and the simple rehabilitation training are effective in the treatment of post-stroke SHS, and promote the status of nailfold microcirculation, the efficacy of the combined therapy is better than that of the latter.</p>

Transcriptional activation of insulin-like growth factor binding protein 6 by 17beta-estradiol in SaOS-2 cells

Osteoblasts can synthesize the insulin-like growth factors (IGFs) and the IGF-binding proteins (IGFBPs), which may either enhance or attenuate IGF-stimulated bone cell proliferation. Since estrogen induced osteoblastic differentiation and proliferation through an estrogen-responsive gene in target cells, we investigated the effects of estrogen on IGFBP-6 expression in the human osteoblastic-like cell line SaOS-2. Expressions of IGFBP-6 protein and mRNA increased 2.8 and 2-fold, respectively, in the presence of 17-beta-estradiol (E2) (0.01 to 1 micrometer) and estrogen receptor (ER) in SaOS-2 cells. On the other hand, E2 induced a 2-fold increase in SaOS-2 cell proliferation. To identify genomic sequences associated with estrogen responsiveness, the 5'-promoter region (-44 to +118) of the IGFBP-6 gene was cloned into a chloramphenicol acetyltransferase (CAT) reporter vector. E2 induced a 3-fold increase in CAT activity in SaOS-2 cells transiently transfected with this construct. Identification of the estrogen-responsive element (ERE) [5'-CCTTCA CCTG-3'] (-9 to +1) in this IGFBP-6 gene promoter region was confirmed using electromobility shift assays and deletion analysis. This functional ERE was important for E2-induced trans-activation of the IGFBP-6 gene. These results demonstrate that E2 exhibits a positive effect on IGFBP-6 gene transcription through estrogen-liganded ER binding to the functional ERE in the IGFBP-6 gene promoter in SaOS-2 cells.