[Transoral robotic thyroidectomy via vestibular approach: a retrospective study of 107 cases in a single center].

Objective: To investigate the short-term outcome of transoral robotic thyroidectomy. Methods: This is a retrospective case series study. The clinicopathologic characteristics and postoperative results of 107 patients who underwent transoral robotic thyroidectomies in the Department of Thyroid and Breast Surgery of the 960th Hospital of People's Liberation Army from May 2020 to August 2023 were retrospectively analyzed. There were 12 males and 95 females, with an age of (31.8±9.4) years (range: 11 to 55 years), including 20 benign tumors and 87 thyroid papillary carcinoma. Postoperative follow-up was carried out through returning visit and telephone, mainly to observe the recovery of postoperative complications, cosmetic effects and recurrence results. Results: All transoral robotic thyroidectomy was successfully completed without conversion to open surgery. The tumor size of thyroid papillary carcinoma patients was (5.6±2.7) mm (range: 2 to 15 mm). Furthermore, central cervical lymph node metastasis was found in 45 cases. The number of central cervical lymph nodes retrieved and metastasized (M(IQR)) were 11 (8) (range: 3 to 26) and 1 (3) (range: 0 to 13), respectively. There was no recurrent laryngeal nerve injury and permanent hypoparathyroidism. The transient hypoparathyroidism after surgery was 8 cases. Other complications occurred as follows: postoperative infection (n=1), left submandibular perforation (n=1), skin scald (n=1), and perioral numbness (n=1), oral tear (n=2). The postoperative stay was 6 (2) days (range: 3 to 11 days). No local lymph node recurrence or metastasis occurred after a follow-up of (22.6±10.0) months (range: 1.0 to 37.4 months). All patients were satisfied with the postoperative cosmetic results, the aesthetic effect score was 9.3 (0.2) (range: 8.4 to 9.6) one month after surgery. Conclusion: For highly screened patients with early thyroid cancer, experienced surgeons can perform a transoral robotic thyroidectomy that has excellent cosmetic results.

Efficacy of erector spinae nerve block for pain control after lumbar spinal surgeries: a systematic review and meta-analysis.

OBJECTIVE: The review aimed to examine the evidence on the efficacy of erector spinae nerve block (ESPB) for pain control after lumbar spinal surgeries. MATERIALS AND METHODS: PubMed, CENTRAL, Embase, and Web of Science were examined for published randomized controlled trials (RCTs) assessing ESPB with control for lumbar spinal surgery patients. The primary review outcome was 24-hour total opioid consumption in morphine equivalents. The secondary review outcomes were pain at rest at 4-6 hours, 8-12 hours, 24 hours and 48 hours, first rescue analgesic timing, needing rescue analgesics number, and postoperative nausea and vomiting (PONV). RESULTS: 16 trials were eligible. Total opioid consumption was significantly lower with ESPB as compared to controls (MD: -12.68 95% CI: -18.09, -7.28 I2=99% p<0.00001). Pain scores at 4-6 hours (MD: -1.37 95% CI: -1.98, -0.76 I2=95% p<0.0001), 8-12 hours (MD: -1.18 95% CI:-1.84, -0.52 I2=98% p=0.0004), 24 hours (MD: -0.53 95% CI:-1.03, -0.04 I2=96% p=0.04) and 48 hours (MD: -0.36 95% CI:-0.84, 0.13 I2=88% p=0.15) were significantly lower in the ESPB group. The meta-analysis found that the ESPB group required a significantly longer time for the first analgesic request (MD: 5.26 95% CI: 2.53, 7.99 I2=100% p=0.002), had lower demand for rescue analgesics (OR: 0.12 95% CI: 0.07, 0.21 I2=2% p<0.00001) and fewer incidence of PONV (OR: 0.27 95% CI: 0.15, 0.49 I2=51% p<0.0001). CONCLUSIONS: ESPB can be highly efficacious for postoperative analgesia in lumbar surgery patients. The block has the capability of reducing opioid consumption in the first 24 hours and pain scores up to 48 hours along with a significant reduction in the need for rescue analgesics and PONV.

[Complications of Da Vinci robot thyroid surgery by bilateral axillo-breast approach].

Objective: To evaluate the complications of Da Vinci robotic thyroid surgery by bilateral axillo-breast approach. Methods: A retrospective analysis of complications was conducted on 1, 198 cases of Da Vinci robotic thyroid surgery by bilateral axillo-breast approach of the 960 th Hospital of the People's Liberation Army from February 2014 to March 2020. There were 263 men and 935 women, age ranged from 9 to 68 years old, and included 288 benign lesions and 910 malignancies according to preoperative imaging examination, FNAC, and intraoperative frozen pathology. Results: Surgical complications occurred in 187 (15.61%) patients, including 10 cases of temporary larynx nerve injury (0.83%), 1 case of permanent larynx nerve injury (0.08%), and 152 cases of temporary hypoparathyroidism (12.69%), no permanent hypoparathyroidism, 1 case of hypoglossal injury (0.08%), 2 cases of facial nerve jaw branch damage (0.17%), 2 cases of trachea injury (0.17%), no esophagus damage, 5 cases of celiac leakage (0.42%), 3 cases of neck skin adhesion (0.25%), 2 cases of subdermal bleeding (0.17%), 2 cases of skin burns (0.17%), 5 cases of hematoma (0.42%), 1 case of cephalic artery rupture (0.08%), 1 case of jugular vein rupture (0.08%), no tumor cultivation, no arm plex nerve, accessory nerve or phrenic nerve damage. Conclusion: Da Vinci robot thyroid surgery by bilateral axillo-breast approach is safe, with less severe complications.

[Mechanism of lncRNA-SRLR induced invasion and metastasis in U2OS osteosarcoma cells].

Objective: To explore the potential mechanism of sorafenib resistance associated long non-coding RNA (lncRNA-SRLR) promoted invasion and metastasis in U2OS osteosarcoma cells. Methods: We transfected U2OS cells with negative control lentivirus (LV-NC) or lncRNA-SRLR overexpressed lentivirus (LV-over/SRLR) particles. LV-NC and LV-over/SRLR stable transfected cells (U20S/NC and U20S/SRLR) were selected by primary cell culture medium containing puromycin. The mRNA expressions of lncRNA-SRLR and procollagen-lysine, procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The effect of lncRNA-SRLR on the invasion of U2OS cells were determined by wound-healing assay and Transwell migration assay. The effect of SRLR on the interleukin-6 (IL-6) secretion of U2OS cells was evaluated by enzyme-linked immunosorbent assay (ELISA) analysis. The subcellular distribution of SRLR in U2OS cells was detected by fluorescence in situ hybridization (FISH) analysis.The expression of PLOD2 in cells was detected by immunofluorescence (IF). The expressions of PLOD2 and focal adhesion kinase (FAK)/signal transducer and activator of transcription 3 (STAT3) signal pathway related proteins in U2OS/NC and U2OS/SRLR cells were detected by western blotting. Results: qRT-PCR assay showed that mRNA expressions of lncRNA-SRLR and PLOD2 in U2OS/SRLR cells were (3 964.97±0.05) and (2.77±0.11), respectively, significantly higher than those in U2OS/NC cells (P<0.001 or P<0.01). The results of wound-healing and Transwell migration assay showed that over-expression of SRLR markedly promoted the invasion ability of U2OS cells (P<0.05). The result of ELISA analysis showed that the IL-6 secretions in U2OS/NC or U2OS/SRLR cells were (125.38±11.22) pg/ml or (119.97±13.43) pg/ml, without statistical significance (P>0.05). The subcellular distribution assay revealed that lncRNA-SRLR is predominately located in the nucleus. The result of IF showed that compared with U2OS/NC cells, the expression of PLOD2 was up-regulated in U2OS/SRLR cells. The result of western blotting showed that over-expression of SRLR significantly increased the expression levels of PLOD2, phosphorylation (p)-FAK and p-STAT3 in U2OS cells (P<0.01). Conclusion: lncRNA-SRLR promotes invasion and metastasis of osteosarcoma by activating PLOD2-FAK/STAT3 signal axis.

[Experimental study on the improvement of Ilizarov transverse tibial bone transport and microcirculation reconstruction technique].

Objective: To observe the effect of traditional and modified Ilizarov transverse tibial bone transport on microvascular regeneration of lower limbs in dogs. Methods: After general anesthesia on 10 experimental dogs, traditional and modified transverse tibial bone transport were performed on both tibias respectively. The control group was treated with the traditional method (the periosteum and bone flap were completely isolated), while the experimental group was treated with the modified method (the fibular periosteum of the open window bone flap was retained). All the external fixators were pulled outwards at a speed of 1 mm every day from 5 days after operation;after one week, the external fixators were moved back every 3 days for one week. The situation of wounds and activity of lower limbs were observed. Simultaneously, the angiogenesis was observed by digital subtraction angiography (DSA) through femoral artery at different stages, and the density of vascular endothelial cells measured by local tissue sections. The data before and after the operation were compared with paired t test. Results: The operation was successfully completed in 10 experimental dogs, and all wounds healed about 1 week after the operation. There was no significant abnormality in lower limb movements in all dogs. Peripheral blood vessel area in middle leg of lower limb in 4 weeks and 8 weeks after operation was (5.9±0.4) mm(2) and (6.9±0.6) mm(2) in control group and it was (6.2±0.6) mm(2) and (8.0±0.6) mm(2) in experimental group, respectively; all were significantly improved than those before the operation ((5.0±0.4) mm(2), (4.9±0.4) mm(2), respectively) (F=446.457, 829.192, both P<0.05). There was no significant differences in vessel area between the two groups at the 4th week after operation (t=1.216, P=0.240), but there was significant difference at the 8th week after operation between the two groups (t=4.423, P=0.000). The percentage of vascular endothelial cells in stained cells under endoscopy was 4.42%±0.28% and 5.63%±0.53% in the control group at the 8th week; and in the experimental group, it was 5.35%±0.26% and 7.18%±0.25%, respectively;all were significantly elevated than those before the operation; and there were significant differences between the two groups (t=7.35, 8.30, both P<0.05). Conclusion: Transverse tibial bone transport and microvascular network regeneration technology can reconstruct the microvasculature below the calf of dogs; the method of window-opening osteotomy is improved to "door" type window-opening, it can retain the lateral periosteum of tibial crest and regenerate the microvasculature network more abundantly.

Potential roles of lncRNA-Cox2 and EGR1 in regulating epidural fibrosis following laminectomy.

OBJECTIVE: Epidural fibrosis, one of the common complications after spinal surgery, seriously affects the surgical decompression effect. Effectively inhibiting the fibrous tissue hyperplasia is pivotal to reduce the scar adhesion. Previous studies showed that early growth response 1 (EGR1) is associated with the fibroblast reactivity induced by transforming growth factor-beta (TGF-ß) and plays a vital regulatory role in scar formation; however, the upstream targets and mechanisms still remain unclear. In this work, it was found that the level of long non-coding ribonucleic acid (lncRNA)-cyclooxygenase-2 (COX2) was significantly negatively correlated with EGR1 expression and the severity of the scar. Therefore, it was conjectured that lncRNA-COX2 may decrease fibroplasia and scar formation by negatively regulating EGR1. MATERIALS AND METHODS: TGF-ß was used to activate the embryonic and adult rat fibroblasts. Rats underwent laminectomy to establish the epidural fibrosis model. The changes in the levels of fibroplasia-related genes were measured and analyzed through messenger RNA (mRNA), lncRNA, and micro RNA expression profile chips. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was applied to determine the levels of EGR1 and lncRNA-COX2, and Western blotting was adopted to detect the content of EGR1, collagen I (Col-1), Col-3, and alpha-smooth muscle actin (α-SMA). The scar formation was reflected by hematoxylin and eosin (HE) staining and Masson staining, and the expression level of α-SMA in the scar tissues was measured via immunohistochemistry. Finally, micro-magnetic resonance imaging (MRI) was utilized to examine the different degrees of epidural fibroplasia. RESULTS: It was found that the reactivity of embryonic rat fibroblasts to the TGF-ß stimulation was different from that of adult rat fibroblasts. LncRNA-COX2 was highly expressed in the embryonic rat fibroblasts, but lowly expressed in the adult rat fibroblasts, which had negative correlations with the EGR1 level in embryonic and adult rat fibroblasts. In addition, it was revealed that the expression of EGR1 in the adult rat fibroblasts was remarkably higher than that in the embryonic rat fibroblasts after the activation with TGF-ß. Meanwhile, the level of lncRNA-COX2 was lowered after the activation, especially in the adult rat fibroblasts. It was discovered in the in-vivo model that the degree of fibroplasia was positively associated with EGR1 level and negatively correlated with lncRNA-COX2 level. CONCLUSIONS: The results of this research elucidated that the down-regulation of lncRNA-COX2 is involved in the epidural scar formation and related to the elevated EGR1 level which regulates the activation of fibroblasts and secretion of massive extracellular matrixes, suggesting that lncRNA-COX2 may modulate the role of fibroblasts in scar formation as an upstream action target of EGR1.

[Upregulation of PLOD2 promotes invasion and metastasis of osteosarcoma cells].

Objective: To investigate the relationship of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods: The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT-PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan-Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U-2OS cells with LV-vector, LV-over/PLOD2, sh-NC and sh-PLOD2. The expression of PLOD2 was detected by qRT-PCR. The impact of POLD2 on U-2OS cell invasion was determined by wound-healing assay and Transwell migration assay. The expressions of PLOD2/FAK/JAK2-STAT3 signal pathway related proteins were detected by western blotting. Results: The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues (P<0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P<0.01). The same results were also observed in qRT-PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 (P<0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+ 9.63)%, significantly higher than (9.67±1.28)% in tissue with low expression of PLOD2 (P<0.001). The result of wound-healing and Transwell migration assay showed that over-expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U-2OS cells (both P<0.01). The result of western blotting showed that over-expression of PLOD2 significantly increased the expression levels of p-FAK, p-JAK2, p-STAT3, but knockdown PLOD2 decreased the levels of p-FAK, p-JAK2, p-STAT3 in U-2OS cells. Conclusions: Up-regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK/JAK2-STAT3 signal pathway.

miR-940 promotes spinal cord injury recovery by inhibiting TLR4/NF-κB pathway-mediated inflammation.

OBJECTIVE: The aim of this study was to investigate the effects of miR-940 and Toll-like receptor 4/Nuclear Factor κB (TLR4/NF-κB) pathways on inflammatory responses and spinal cord injury (SCI). MATERIALS AND METHODS: This study first established a model of spinal cord injury in mice. The grip force measurement was used to detect the recovery of the forelimb, left forelimb and right forelimb of SCI mice. The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-940 and macrophage receptor TLR4 in SCI mice. In addition, the protein levels of TLR4 and inducible nitric oxide synthase (iNOS) in SCI mice were detected by Western blot. MiR-940 mimic was injected into the injured area of SCI mice to explore the effect of miR-940 overexpression on TLR4 and myeloperoxidase (MPO) expression as well as the protein levels of TLR4, P65 and iNOS. Furthermore, the grip strength of SCI mice with double forelimb, left forelimb and right forelimb was detected by the grip force test after miR-940 overexpression. RESULTS: Compared with the sham-operated mice, the grip strength of the forelimb, left forelimb, and right forelimb of the SCI group showed significant obstacles. Meanwhile, the expression of miR-940 was remarkably decreased in SCI mice along with significant elevation of the inflammatory response-related factors including TRL4 and iNOS. Then we injected SCI mice with miR-940 mimics into the spinal cord injury area and found that miR-940 overexpression decreased the expression levels of TLR4 and MPO. At the same time, the overexpression of miR-940 markedly decreased the protein levels of TLR4, P65, and iNOS in SCI mice. In addition, miR-940 overexpression improved the grip strength of the left and right forepaws and the simultaneous grip strength of the two claws of the SCI mice than those of the simple injury group. CONCLUSIONS: High expression of miR-940 can promote the recovery of spinal cord injury by downregulating the TLR4/NF-κB signaling pathway and inhibiting inflammation.

[Effects of menopausal factor on fine anatomy of bladder urethra and vagina in women undergoing vaginal delivery].

Objective: To explore the effects of menopausal factor on fine anatomy of bladder, urethra and vagina in women undergoing vaginal delivery. Methods: Gynecological patients in Nanfang hospital from January 2013 to October 2016 were collected, and then the patients whose MRI images quality meet the require of reconstruction, with the history of vaginal delivery experience, without any cesarean section experience, and the first labor time was ≤30 years old were enrolled. The patients who had pelvic floor dysfunction when done MRI examinations were excluded. Finally, 238 cases were randomly selected out, and 238 models of data were reconstructed and measured by Mimics and UG software. The independent t test was used to do the comparison between menopausal group and those not yet menopause. Results: First, we built 238 3D models totally. Second, the parameters related with bladder and urethra: the angle of bladder and urethra, ß angle, urethra pubic angle, α angle, retropubic space, the length between bladder neck and edge of pubic midpoint, and urethral striated muscle thickness of menopausal group were bigger than those of pre-menopausal group. While for the urethra tilt angle, the former group was smaller than that of the latter group. But there was no significant statistical differences between two groups. Third, the parameters related with vagina: the proximal urethral vaginal gap of the post-menopause group was smaller than that of pre-menopausal group, while for the middle and distal urethral vaginal gap, the former group was bigger than that of the latter group. The length and width of vaginal anterior wall of the post-menopausal group were smaller than those of pre-menopausal group. Beside the middle urethral vaginal gap, all the difference between two groups had no statistical meaning. The 2D shape of axial vaginal, H type occupation of the former group was obviously lower than that of the latter group. Conclusions: Menopause has an effect on fine anatomy of pelvic organ. Especially on the shape of vagina, the middle urethral vaginal gap become much wider after menopause, the occupation of shallow concave type become much higher post-menopausal. It means the lateral and backward support function of the urethra and vagina is relatively weakened after menopausal, and the shape of vagina become smaller.

Comparative study between mini-open TLIF via Wiltse's approach and conventional open TLIF in lumbar degenerative diseases.

OBJECTIVE: To compare the clinical effects, operation safety and radiation exposure of mini-open TLIF via Wiltse's approach (MOTLIF) and conventional open TLIF (COTLIF) in the treatment of single-segment lumbar degenerative disease via the prospective control study. PATIENTS AND METHODS: A total of 77 patients were enrolled from November 2012 to July 2014, including 42 patients in the mini-open group (MOTLIF) and 35 patients in the COTLIF group. Oswestry Disability Index (ODI) and Visual Analogue Scale (VAS) scores before operation, operation time, intraoperative blood loss, postoperative drainage volume, blood transfusion rates, postoperative bedridden time, postoperative hospital stays, intraoperative fluoroscopic time, levels of serum creatine phosphokinase (CPK) before operation, 3 days and 1 week after operation, VAS scores before operation, 3 days and 1 week after operation, and ODI and VAS scores in the last follow-up between the two groups were compared. RESULTS: There were significant differences between the two groups in the operation time, intraoperative blood loss, postoperative drainage, blood transfusion rates, postoperative bedridden time, postoperative hospital stays and intraoperative fluoroscopic time; all indicators in MOTLIF group were superior to those in COTLIF group (p<0.05). There were no significant differences between the two groups in levels of serum CPK before operation and 1 week after operation (p>0.05). However, 3 days after operation, the level of serum CPK in COTLIF group was increased more significantly than that in MOTLIF group (647.4±178.6 vs. 467.4±189.4). There were no differences between the two groups in ODI and VAS scores before operation; ODI score in MOTLIF group in the last follow-up was significantly superior to that in COTLIF group (p>0.05). And VAS scores at 3 days and 1 week after operation and the last follow-up in MOTLIF group were superior to those in COTLIF group (p<0.05). CONCLUSIONS: Compared with the conventional open TLIF, mini-open TLIF via Wiltse's approach using the self-designed operating apparatus is characterized by the convenient operation, small trauma and quick recovery after operation. At the same time, the radiation exposure is lower and long-term follow-up effect is superior. Its short-term and long-term effects in the treatment of lower lumbar degenerative disease are also superior.

Biomechanical strength impact of lateral wall breach on spinal pedicle screw fixation.

OBJECTIVE: The purpose of our study was to make a comparison between the fixation strength of optimum placed pedicle screw (OS) and re-directionally accurate placed pedicle screw (RS) after lateral pedicle breach. PATIENTS AND METHODS: A total of 30 fresh lumbar vertebrae (L1-5) were gained from 6 male or female pigs weighing about 100 kg, which were divided into 2 groups according to different ways of pedicle screws placement: OS group (n=30) and RS group (n=30). MTS machine was employed to detect the screw loosening and axial pullout. We examined seating torque, screw-loosening force, the maximal torque and post-loosening axial pullout in each pedicle screw. RESULTS: Maximal insertion torque of OS was (111.6±8.4) N•cm and RS was (79.0±6.3) N•cm, which indicated a significant difference (Z=3.012, p=0.003). Seating torque of OS and RS were (85.9±5.6) N•cm and (60.3±4.8) N•cm separately, and the difference was statistically significant (Z=2.799, p=0.006). Screw loosening force of OS and RS were (75.9±7.0) N and (52.4±6.3) N respectively, and the difference was statistically significant (Z=2.652, p=0.003). Post-loosening axial pullout force of OS and RS were (328.5±11.3) N and (269.1±9.6) N separately, demonstrating that the difference was statistically significant (Z=2.865, p=0.004). CONCLUSIONS: RS placement is an alternative for remediation following a lateral wall breach evidenced by significantly decreased seating torque, screw loosening force, the maximal torque and post-loosening axial pullout compared with OS.

A multi-level drill guide template improves the accuracy of pedicle screw placement in lumbar spine.

OBJECTIVE: Free-hand technique is widely used in pedicle screw placement for lumbar spine and generally safe; however, screw malposition still occurs. To develop a novel multi-level drill guide template for pedicle screw placement in lumbar spine and evaluate its accuracy. MATERIALS AND METHODS: Twelve lumbar cadaveric specimens were randomly allocated into guide template group (n=6) and free-hand group (n=6). Computed tomography (CT) scans were obtained for reconstruction of three-dimensional (3D) model of each lumbar vertebra, and further an individual guide template was designed. Then the templates and their corresponding vertebra were developed by rapid prototyping (RP) technology. With the guide of the templates, screws were inserted via mini-open Wiltse approach. The positions of the screws were assessed based on postoperative CT images. RESULTS: In total, 120 pedicle screws inserted (guide template group: n=60 vs. free-hand group: n=60). For all 30 vertebras in the guide template group, all pre-designed personalized drill guide templates can be fitted into the facet joints of each vertebra well. Furthermore, our results revealed a significant improvement for the guide template group in the accuracy rate (p=0.026). CONCLUSIONS: Armed with advantages of minimal invasion, enhanced accuracy and safety, the novel technique of multi-level drill guide template can be properly applied in pedicle screw placement for lumbar spine and promises to be a potential option in clinical application.

[Value of the concentration and integrity of serum cell-free DNA for the clinical diagnosis of esophageal carcinoma].

Objective: To explore the diagnostic value of serum cell-free DNA (cfDNA) concentration and integrity for esophageal carcinoma. Methods: Venous blood samples from 68 patients with esophageal cancer, 36 patients with benign esophageal lesions and 45 healthy subjects were collected. Circulating cfDNA was verified through quantitative real-time PCR (Alu-qPCR) using Alu-115 and Alu-247 primers. DNA integrity index was calculated as the ratio of Alu-qPCR results (Alu247/115). Concentrations of carcino-embryonic antigen (CEA) and squamous cell carcinoma associated antigen (SCC) were detected by chemiluminescence analyzer assay. Statistical analysis was performed using Mann-Whitney U test, Kruskal-Wallis H test and Spearman correlation test. The Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of each index to esophageal carcinoma. Results: The median absolute serum Alu115 and the Alu247/115 index (1 162.0 ng/ml, 0.57) in esophageal cancer group were significantly higher than those in benign esophageal disease group (496.7 ng/ml, 0.43) and in healthy control group (432.3 ng/ml, 0.42) (all P<0.01, respectively). The Alu115 and Alu247/115 index of serum DNA in benign esophageal disease group were no statistically different from those in the healthy control group (all P>0.05, respectively). The levels of cfDNA and its integrity were not significantly correlated with age, gender, tumor differentiation, or disease stage according to American Joint Committee on Cancer (AJCC) staging system in the esophageal cancer group (all P>0.05). The serum Alu247/115 index of Stage Ⅲ patients was higher than that of Stage Ⅰ~Ⅱ patients(P<0.05). The serum Alu247/115 index of Stage Ⅳ was higher than that of Stage Ⅲ(P<0.05). In the esophageal cancer group, both of serum Alu115 and Alu247/115 index had no correlation with CEA or SCC (all P>0.05). The area under the ROC curve (AUC) of Alu115 and Alu247/115 index were 0.867 and 0.854, respectively, which were both higher than that of CEA (0.622) and SCC (0.753). The addition of Alu115 or Alu247/115 index to CEA and SCC detection increased the sensitivity of the diagnosis of esophageal cancer by 95.6% and 94.1%, respectively. Conclusions: The detection of serum cfDNA concentration and integrity is helpful to the early diagnosis and monitoring of esophageal cancer. Their diagnostic value of esophageal cancer is better than that of the traditional tumor markers CEA and SCC.

Maternal pre-pregnancy body mass index, gestational weight gain influence birth weight.

BACKGROUND: Evidence suggests that pre-pregnancy body mass index and gestational weight gain have impact on pregnancy and birth weight, yet whether maternal gestational weight gain has a differential effect on the rates of adverse birth weight among women with different pre-pregnancy body mass index categories are unknown. METHODS: We selected 1617 children matched with their mothers as study subjects. The subjects were divided into three categories: weight gain below the American Institute of Medicine guidelines, weight gain within the American Institute of Medicine guidelines and weight gain above the American Institute of Medicine guidelines. RESULTS: The prevalence of pre-pregnancy underweight and overweight/obese women was 16.3% and 12.3%. And nearly 15.2% of the women had gestational weight gain below American Institute of Medicine guideline, 52.1% of the women had gestational weight gain above American Institute of Medicine guideline. Maternal overweight and obese was associated with increased risk for macrosomia and large-for-gestational age. Women had gestational weight gain below American Institute of Medicine guideline were more likely to have low birth weight and small-for-gestational age than women who had gestational weight gain within American Institute of Medicine guideline. Furthermore, the risks for macrosomia and large-for-gestational age were increased in women with above American Institute of Medicine guideline. And for women with a normal weight before pregnancy, gestational weight gain above the American Institute of Medicine guidelines were associated with higher rates of macrosomia and large-for-gestational age, compared with the women of similar pre-pregnancy weight category but with gestational weight gain within the American Institute of Medicine guidelines. CONCLUSIONS: Women with abnormal pre-pregnancy body mass index and gestational weight gain are at risk for adverse birth weight outcomes. Moreover, gestational weight gain has a differential effect on the rates of adverse birth weight outcomes between women of different pre-pregnancy body mass index categories.

Functions of microRNAs in osteoporosis.

Osteoporosis (OP) is a kind of disease with a 25% incidence, characterized by the bone mass loss, bone microstructure damage, increased bone fragility, and easy fracture. miRNA plays an important regulatory role in the process of bone remodeling, especially in the regulation of differentiation and function of osteoblasts and osteoclasts, and the development and progression of OP and other bone diseases. In the future, it is expected to delay the bone loss and promote the bone remodeling via the overexpression or inhibition of specific miRNAs in specific tissues, thereby treating OP.

Inverse incoherent engineering.

We propose a protocol to construct shortcuts of adiabaticity for open quantum systems in unitary evolution. Using the dynamical invariants of open quantum systems, we design a convenient form of the driver Hamiltonian to accelerate the adiabatic decoherence free subspaces scheme (TDFs) and engineer a quantum state from the initial state into the target state. Since the trajectory of TDFSs is determined by the incoherent control process, we would like to call it as the inverse incoherent engineering protocol. We apply the method to a two-qubits system which interacts with a time-dependent vacuum squeezed field to prepare some maximally entangled states of it. The results illustrate that our protocol can be used both in the adiabatic and nonadiabatic regime with perfect fidelity.

[Investigation of influencing factors associated with neck-shoulder symptoms among middle school students in three cities in China].

Objective: The aim of this study was to describe the distribution of neck-shoulder symptoms among middle school students, and to explore its influence factors. Methods: A cross-sectional survey was conducted on 24 middle schools in Shenyang, Zhengzhou and Shenzhen by purposive sampling method. In each middle school, 3 to 4 classes were selected from each grade all the students in the selected class would be recruited to the survey to investigate the demographic characteristics, neck-shoulder symptoms, physical exercise time, academic stress, screen behavior, sedentary behavior and other information by questionnaire. A total of 10 566 questionnaires were issued and 10 270 valid questionnaires were withdrawn. The prevalence of neck-shoulder symptoms among students were compared by different characteristics. Logistic regression models were applied to examine influencing factors associated with neck-shoulder symptoms. Results: The prevalence of neck-shoulder symptoms among middle school students was 19.2% (1 968/10 270), while it was 22.6% (1 137/5 039) among girls and 15.9% (831/5 231) among boys; the difference showed statistical significance (P<0.001). The prevalence of neck-shoulder symptoms was separately 13.3% (253/1 901) in seventh grade, 16.8% (326/1 942) in eighth grade, 18.5% (299/1 617) in ninth grade, 21.8% (417/1 915) in sophomore, 21.4% (345/1 611) in junior, 25.5% (328/1 284) in senior; the difference showed statistical significance (P<0.001). The multivariate logistic regression analysis showed that in the last 3 months, students who always playing mobile phone ≥40 min continuously (OR=4.66, 95%CI: 3.95-5.49), watching TV ≥40 min continuously (OR=4.01, 95%CI: 3.39-4.73), using computer ≥40 min continuously (OR=3.61, 95%CI: 3.09-4.23), doing homework ≥60 min continuously (OR=3.25, 95%CI: 2.79-3.79), the average daily sitting time ≥10 h (OR=4.95, 95%CI: 4.25-5.77), and always sitting ≥90 min continuously (OR=5.18, 95%CI: 4.42-6.06) were risk factors of neck-shoulder symptoms. Conclusion: The prevalence of neck-shoulder symptoms was high among middle school students in China, especially girls in senior grades. Long time, high frequency video behaviors and sedentary behaviors were related to the occurrence of neck-shoulder symptoms among middle school students.

[Mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1 induced invasion and metastasis of esophageal cancer cell EC-109].

Objective: To investigate the effect and mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1, (LncRNA-MALAT1) on invasion and metastasis of esophageal cancer cell EC-109. Methods: EC-109 cells were transfected with lentiviral vector carrying short hairpin RNA of MALAT1( shRNA-MALAT1) or a nonspecific shRNA control (shRNA-control). The expressions of MALAT1, microRNA-200a, ZEB1 and ZEB2 were detected by qRT-PCR. The effect of shRNA-MALAT1 on invasion of EC-109 cells was determined by transwell assay. The expressions of components of epithelial-msenchymal transition pathway in EC-109 cells were determined by immunofluorescence array and western blotting. The expression relationship between MALAT1 and miR-200a in EC-109 cells was detected by dual-luciferase reporter assay. Results: The result of qRT-PCR showed that the expressions levels of MALAT1, ZEB1 and ZEB2 in shRNA-MALAT1 group were 0.43±0.06, 0.64±0.04 and 0.51±0.04, respectively, significantly lower than 0.97±0.08, 1.06±0.07 and 0.98±0.05 in shRNA-control group and 1 in control group, respectively(all P<0.05). Transwell assay showed that the number of invaded cells in shRNA MALAT1 group was (96.81±10.43) per low-power field, markedly lower than that of (278.44±13.28) per low-power field in shRNA-control group (P<0.01). Immunofluorescence staining and Western blotting showed that MALAT1 downregulation significantly reduced the expressions of proteins related to EMT signal pathway in EC-109 cells.Dual luciferase reporter assay showed that compared to negative control, the activities of luciferase reporter in EC-109 cells co-transfected with pmirGLO-MALAT1-wt and miR-200a were significantly down-regulated. While co-transfected pmirGLO-MALAT1-mut with miR-200a mimics had no effect on the luciferase reporter activities of MALAT1. Conclusion: LncRNA MALAT1 functions as a competing endogenous RNA to regulate the expressions of ZEB1 and ZEB2 by sponging miR-200a and promotes invasion and migration of esophageal cancer cells through inducing epithelial-mesenchymal transition.

[Expression of long non-coding RNA MALAT1 in osteosarcoma and its effect on invasiveness and metastatic potential of osteosarcoma cells].

OBJECTIVE: To investigate the significance of long non-coding RNA MALAT1 expression in osteosarcoma, and the potential mechanism by which MALAT1 promotes tumor metastasis. METHODS: Twenty cases of osteosarcoma in the First Affiliated Hospital of Xinxiang Medical University and Ping Ding Shan First People's Hospital were collected from January 2014 to December 2015. The expression of MALAT1 in osteosarcoma tissue and paired adjacent noncancerous tissue were analyzed by qRT-PCR. Correlation of MALAT1 expression in osteosarcoma with clinical pathologic features was performed by the Mann-Whitney U test. U-2OS cells were transfected with lenti-virus carrying MALAT1-shRNA and nonspecific shRNA (LV-vector). The expression of MALAT1 was detected by qRT-PCR. The cell activity was evaluated by MTT asssy. The impact of MALAT1-shRNA on invasion in U-2OS cells were determined by transwell migration assay. The expression of Wnt/ß-catenin signal pathway related proteins were detected by Immunofluorescence stain and Western blot. RESULTS: The expression level of MALAT1 in osteosarcoma tissue was higher than that in paired adjacent noncancerous tissue and correlated significantly with nodal and pulmonary metastasis(P<0.01). MTT assay showed that knockdown of MALAT1 with lenti virus-MALAT1 shRNA inhibited the growth of U-2OS cells, along with marked decrease of invasive ability of U-2OS cells in the transwell migration assay. By immunofluorescence stain and Western blot assay, MALAT1 significantly reduced the expression of ß-catenin, MMP7, and c-MYC in U-2OS cells. CONCLUSIONS: The expression of MALAT1 is high in osteosarcoma and correlates with tumor metastasis. MALAT1 promotes invasion and metastasis of osteosarcoma cells likely thought the Wnt/ß-catenin signal pathway.

Construction of a protein-protein interaction network of Wilms' tumor and pathway prediction of molecular complexes.

Wilms' tumor (WT), or nephroblastoma, is the most common malignant renal cancer that affects the pediatric population. Great progress has been achieved in the treatment of WT, but it cannot be cured at present. Nonetheless, a protein-protein interaction network of WT should provide some new ideas and methods. The purpose of this study was to analyze the protein-protein interaction network of WT. We screened the confirmed disease-related genes using the Online Mendelian Inheritance in Man database, created a protein-protein interaction network based on biological function in the Cytoscape software, and detected molecular complexes and relevant pathways that may be included in the network. The results showed that the protein-protein interaction network of WT contains 654 nodes, 1544 edges, and 5 molecular complexes. Among them, complex 1 is predicted to be related to the Jak-STAT signaling pathway, regulation of hematopoiesis by cytokines, cytokine-cytokine receptor interaction, cytokine and inflammatory responses, and hematopoietic cell lineage pathways. Molecular complex 4 shows a correlation of WT with colorectal cancer and the ErbB signaling pathway. The proposed method can provide the bioinformatic foundation for further elucidation of the mechanisms of WT development.