Objective To explore the changes of bone metabolic markers during early stage of preterm infants, as well as the relationship with their nutrition status. Methods Preterm infants with gestational age 30-35 weeks admitted to our Hospital were collected from November 2012 to April 2013. Venous blood samples obtained within 24 hours after birth and between 8:00-9:00 AM two weeks after birth were used to determine the Serum β-CTx, osteocalcin ( OC) and propeptide of type I procollagen (PINP) levels by Electro-Chemiluminescence. Analysis of changes of these bone metabolic markers and their relationship with early stage nutrition related indicators were also performed. Results A total of 60 premature infants were collected. There was no significant correlation among serum β-CTx, OC and PINP within 24 hours after birth ( r=0. 170, P＞0. 05 ) . The Serum β-CTx within 24 hours after birth was negatively correlated with gestational age (r= -0. 603, P＜0. 05), whereas the serum OC within 24 hours after birth was positively correlated with gestational age ( r=0. 581, P＜0. 05 ) . However, PINP wasn′t correlated with gestational age significantly (r=0. 134,P＞0. 05). Serumβ-CTx, OC and PINP at 2 weeks after birth increased significantly from the baseline level detected within 24 hours after birth ( P＜0. 05 ) .Δβ-CTx was positively correlated with ΔOC (r=0. 600,P＜0. 05). There was no significant correlation between ΔPINP and Δβ-CTx (r=0. 045,P＞0. 05), as well as ΔOC and ΔPINP (r=0. 110,P＞0. 05).ΔOC was positively correlated with average daily calorie ( P＜0. 05 ) and average daily P/E ( P＜0. 05 ) , negatively correlated with cumulative loss of caloric ( P＜0. 05 ) . There was no significant correlation between Δβ-CTx or ΔPINP with nutrition related indicators of this study. Conclusions Serum OC within 24 hours after birth of preterm infants and their gestational age are positively correlated, while β-CTx detected at the same time and gestational age are negatively correlated. Vigorous metabolism of premature bone occurs during the first two weeks after birth, as the serum β-CTx, OC and PINP levels increased significantly. We suggest that reasonable calorie intake and appropriate protein calorie ratio at early stage after birth is very important for the development of bone of preterm infants.

To investigate the effects of ATRA, acitretin and tazarotene on the growth and apoptosis of human tongue squamous cell carcinoma cell line Tca8113. The effect of retinoids on growth of Tca8113 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis with double staining with Annexin V-FITC and PI, and active caspase-3 analysis with the staining of FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. Streptavidin-biotin complex (SABC) immunocytochemical assays were employed for the detections of Bax/Bcl-2 proteins expressions. Our results showed that the retinoids inhibited growth of Tca8113 cells in a dose- and time-dependent manner with maximal inhibition 24 h after treatment of 10-5 mol/L. 10-5 mol/L retinoids altered cell cycle distribution of Tca8113 cells, revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. 10-5 mol/L retinoids significantly induced apoptosis of Tca8113 cells (all P<0.05), elevated the cells population with detectable active caspase-3 (P<0.05 for all), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (all P<0.05). Acitretin played a most prominent role among the retinoids. It is concluded that the inhibition of cell cycle progress of Tca8113 cells by ATRA, acitretin and tazarotene is one of the possible mechanisms for proliferation arrest of Tca8113 cells elicited by the retinoids. The retinoids mediate apoptosis in Tca8113 cells that may be caspase-dependent through mitochondria pathway. High concentration retinoids inhibit growth of Tca8113 cells in vitro by interfering with proliferation and inducing apoptosis of cells. Acitretin may be an alternative medicine for the prevention and treatment of tongue squamous cell carcinoma.

Objective To probe the relationship between the expression of TL1A and the level of IFN-γ secreted by T cells in the acute stage of Guillain-Barre syndrome (GBS). Methods ① Six-week female Bal b/c mice were immunized by purified recombinant human soluble TNF-like molecular 1A (rhsTL1A) protein. The polyclonal antibody against rhsTL1A was identified by immunofluorescence using human umbilical vein epithelial cells (HUVEC). ② To detect the biologic activity of rhsTL1A, the peripheral blood mononuclear cells (PBMC) from the healthy donors were separated by Ficoll gradient centrifugation and were seeded on 96-well plates with medium containing 2 μg/ml PHA (control group), 2 μg/ml PHA + 25 ng/ml rhsTL1 A, 2 μg/ml PHA + 100 ng/ml rhsTL1A and 2 μg/ml PHA + 400 ng/ml rhsTLlA respectively. T cell proliferation assay was carried out using ~3H-TdR. ③ IFN-γ productions in the sera of the children with GBS in the acute stage were detected by ELISA. ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage was detected with flow, cytometry. ⑤PBMC from the children in acute GBS were separated and cultured in the environment adding 2 μg/ml PHA and 400 ng/ml rhsTL1A in vitro. Then, the IFN-γ in the supernatant was determined by ELISA kit after 72 hours. Results ① hTL1A A expressed by eukaryotic HUVECs was recognized by rhsTL1 A polyclonal antiserum. ② The result of T cell proliferation assay showed that SI of 25 ng/ml rhTL1A, 100 ng/ml rhTL1A A and 400 ng/ml rhTL1A group was increased compared with control group. The SI of 2 μg/ml PHA +400 ng/ml rhsTL1 A group was the highest (2. 65) among them. ③ IFN-γ productions in the sera of the children with GBS in the acute stage ((102. 25±22. 17) pg/ml) were increased significantly compared with healthy control ((28.75 ± 1.31) pg/ml, t = 3. 309, P < 0. 05). ④ The ratio of CD_3~+ TL1A~+ T cells to CD_3~+ T cells in the peripheral blood of the children with GBS in acute stage (18.22%± 1.83%) was enhanced significantly compared with healthy control (5. 17% ±0. 48%, t = 6. 884, P < 0. 01). ⑤ PBMC both in healthy control and the acute GBS secreted more IFN-γ markedly ((43.56± 4.41) pg/ml and (180.64 ± 38.39) pg/ml) after being incubated in 2 μg/ml PHA and 400 ng/ml rhsTL1A (t =4. 523 and 2. 600, P <0. 01 and 0. 05 respectively). Moreover, PBMC in acute GBS secreted more IFN-γ, than that of the healthy group markedly (t = 3. 545, P < 0. 05). Conclusions ① The mouse antiserum recognizing rhsTL1A is successfully obtained. ② In this study, 400 ng/ml rhsTL1A promotes the proliferation of T cells activated by 2 μg/ml PHA, indicating that rhsTL1A has biological activity. ③ The expression of hTL1A of activated T cells in the peripheral blood of the children with acute GBS is up-regulated. These TL1A proteins promote the secretion of IFN-γ through binding to their receptors DR_3.

AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.

[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.

We propose a clinical pathway of acute exacerbations of chronic obstructive pulmonary disease (AECOPD) based on state machine. Clinical event-driven response was utilized to control workflow execution of the AECOPD clinical pathway. By comparison with the traditional clinical pathway management, clinical numerical results showed that the proposed method was better in hospitalization days, average hospitalization expense and aberration rate, and better handled the variability in the AECOPD clinical pathway execution.
Critical Pathways ; Humans ; Pulmonary Disease, Chronic Obstructive ; diagnosis ; nursing ; therapy

Objective To analyze the clinical value of immature granulocytes in peripheral blood for prediction of persistent systemic inflammatory response syndrome (SIRS) in patients with acute pancreatitis (AP). Methods 1 973 patients with AP in Hunan People's Hospital from 2012 to 2017 were retrospectively enrolled and divided by SIRS duration into the persistent SIRS group, temporary SIRS group and non-SIRS group. The independent risk factor for persistent SIRS in AP patients was evaluated by Logistic regression analysis, and predictive value of immature granulocytes for persistent SIRS in AP patients was analyzed by the receiver operating characteristic (ROC) curve. Results These 1 973 AP patients (1 165 males, 59.0%) with an average age of 49 (40, 60) years old, including 288 persistent SIRS, 189 temporary SIRS and 1 496 non-SIRS cases. There was no significant difference in gender, age and etiology among three groups. Compared with non-SIRS group, more severe symptoms were observed in the temporary and persistent SIRS groups. Moreover, The acute physiology and chronic health evaluation Ⅱ(APACHEⅡ), CT severity index (CTSI), multiple organ failure (MOF) and acute respiratory distress syndrome (ARDS) incidence, mortality and C-reactive protein (CRP), white blood cell count (WBC), procalcitonin (PCT) and immature granulocytes in persistent SIRS group were further higher than those in the temporary SIRS group [APACHEⅡ: 9 (6, 12) vs. 5 (3, 7), CTSI: 6 (4, 6) vs. 4 (3, 6), MOF incidence: 92.0% vs. 32.8%, ARDS incidence: 39.9% vs. 10.1%, morbidity: 11.1% vs. 4.2%, CRP (mg/L): 25.00 (0.80, 212.25) vs. 0.80 (0.80, 123.50), WBC (×109/L): 15.17±6.78 vs. 14.84±5.86, PCT (μg/L): 0.23 (0.10, 1.76) vs. 0.10 (0.10, 0.31), immature granulocytes: 1.95 (0.90, 4.95) % vs. 0.80 (0.40, 2.10) %, all P < 0.05]. Logistic regression analysis showed that besides pancreatic necrosis, WBC and CRP, immature granulocyte was an independent risk factor for persistent SIRS associated with AP [odds ratio (OR) = 1.844, 95% confidence interval (95%CI) = 1.372-2.220]. ROC curve showed that immature granulocytes had better predictive value for persistent SIRS, the area under the curve (AUC) was 0.806, which was significantly higher than the APACHEⅡ (AUC = 0.783), CTSI (AUC = 0.752), PCT (AUC = 0.676), CRP (AUC = 0.677), WBC (AUC = 0.644). The cut-off value of immature granulocyte was 0.65%, the sensitivity was 84.0%, the specificity was 66.3%, the positive predictive value was 62.4%, and the negative predictive value was 76.3%. Conclusion Immature granulocyte in peripheral blood is a potential indicator for persistent SIRS in AP patients.

Objective: To study the learning curve of laparoscopic pacreaticoduodenectomy (LPD) with a view to find an appropriate way to develop LPD step by step. Methods: 112 consecutive patients who completely underwent LPD in a single surgery center at the First People’s Hospital of Changzhou from December 2015 to February 2018 were retrospectively reviewed. By using both the cumulative sum (CUSUM) and the risk-adjusted CUSUM (RA-CUSUM) methods to analyze the perioperative data of these patients, the learning curve of LPD was studied in a more scientific way. Results: The learning curve could be divided into three phases: Phase 1, the initial period (the initial 45 patients); Phase 2, the enhancement period (the subsequent 31 patients); Phase 3, the maturation period (the remaining patients). For these 3 phases, the corresponding operative times were (448.4±75.0), (381.3±74.3), and (336.2±52.1) min, respectively (P<0.05). The intraoperative blood losses were (373.3±250.1), (332.3±211.6), and (265.3±253.2) ml, respectively (P<0.05). The times to oral intake were 6.0(5.0, 8.0), 5.0(3.0, 6.0), and 3.0(3.0, 5.0) days, respectively (P<0.05). The number of lymph nodes harvested were (10.0±7.0), (8.8±4.3), and (13.3±6.2), respectively (P<0.05). All these showed significant improvement through the 3 phases. On the other hand, the postoperative stays, the postoperative pancreatic fistula rates were also decreased. But these failed to reach statistical significance. Vascular reconstruction was carried out in the 48th patient in phase 2 of the study. Conclusions The initial phase of LPD passed after LPD for 46 patients, but the maturation phase occurred after LPD on 76 patients. Vascular reconstruction should be considered as passing through the learning phase rather than reaching the maturity phase. Adjustments made in the enhancement phase helped to get through the maturation phase earlier.

AIM:The paper is to explore a rapid and simple method for the culture of mouse primary thyroid cells.METHODS:Mouse thyroid cells were isolated by enzyme digestion and cultured with improved medium,and their morphology,characteristics and secretory function were observed within 14 d.RESULTS:In the cultures,the active pri-mary cells were obtained from the thyroid tissue after digestion for 25 min;adherent growth was observed on the 2nd day.And secondary follicles appeared from the 5th to 7th day.Over 95%cells were detected with thyroglobulin.The secretion of total triiodothyronine and total thyroxine maintains over 60%in 7 d.The expression levels of specific genes can still maintain more than 50%in 10 d.CONCLUSION:Mouse thyroid primary cells can be rapidly cultured by this method,and the cells can be used for studying thyroid endocrine secretion within 7 d and studying thyroid genes within 10 d.

Objective:To explore the application value of individualized three-dimensional (3D) printing positioning guides in localization and resection of intracranial lesions.Methods:Fifteen patients with intracranial space occupying lesions underwent resection in our hospital from March 2021 to May 2021 were selected in our study. Brain images by CT and MRI as raw data were used to design individual positioning guides. The positioning guides were placed on the patient's skin before resection to mark the location and boundary of the lesions with a marker, and neuro-navigation was used to verify the accuracy. During the resection, the location of the lesions was identified through microscope by the surgeons. Postoperative CT and MRI were used to evaluate the lesion resection.Results:The individualized positioning guides of 15 patients fit the skin well, and the skin incision and bone window were designed to meet the surgical requirements. All surgeries were completed in one time, and the lesion tissues were successfully removed. During the surgeries, the skin incision was not adjusted for secondary expansion. Brain MRI reexamination within 48 h of surgery showed that the lesions of 11 patients with tumors were removed satisfactorily (total resection in 9 and subtotal resection in 2); brain CT reexamination within 12 h showed that the clearance rate of hematomas in 3 patients was above 80% and that in 1 patient was 70%. No patients had cerebrospinal fluid leakage, intracranial hematoma, intracranial infection or other serious complications. All patients recovered well during the 1-3 months of outpatient/telephone follow-up.Conclusion:The positioning method with personalized 3D printing guides is simple and convenient, enjoying accurate positioning results, which can assist the clinicians to optimize the preoperative planning, optimize the surgical incision design, and is worthy of promotion and application in primary hospitals.