Sepsis risk calculator-guided antibiotic management in neonates with suspected early-onset sepsis / 中国当代儿科杂志

OBJECTIVE@#To evaluate the efficacy of sepsis risk calculator (SRC) in guiding antibiotic use in neonates with suspected early-onset sepsis (EOS).@*METHODS@#A total of 284 neonates with a gestational age of ≥ 35 weeks were enrolled as the control group, who were hospitalized in the Children's Hospital of Chongqing Medical University from March to July, 2019 and were suspected of EOS. Their clinical data were retrospectively collected and the use of antibiotics was analyzed based on SRC. A total of 170 neonates with a gestational age of ≥ 35 weeks were enrolled as the study group, who were admitted to the hospital from July to November, 2020 and were suspected of EOS. SRC was used prospectively for risk scoring to assist the decision making of clinical antibiotic management. The two groups were compared in terms of the rate of use of antibiotics, blood culture test rate, clinical outcome, and adherence to the use of SRC.@*RESULTS@#Compared with the control group, the study group had a significantly higher SRC score at birth and on admission (@*CONCLUSIONS@#The use of SRC reduces the rate of empirical use of antibiotics in neonates with suspected EOS and does not increase the risk of adverse outcomes, and therefore, it holds promise for clinical application.

The observe of clinical effect of treating lumbar intervertebral disc herniation by bone setting manipulation of different directions / 中国骨伤

<p><b>OBJECTIVE</b>To compare the effect between lumbar backwards flexion manipulation and rotating manipulation for treating lumbar intervertebral disc herniation.</p><p><b>METHODS</b>Two hundred and nine patients of lumbar intervertebral disc herniation, male 131, female 78, the age from 20 to 79 years old, 58 cases of all these patients age above 50. According to diagnosis the ladder of the 92 cases bulging type, 69 hernia type, 48 cases free type. The patients were randomly divided into treatment group (107 cases) and control group (102 cases). All the patients were treated with the three-dimensional computer-controlled traction therapeutic apparatus, with continued traction for 30 minutes. After traction, lumbar backwards flexion manipulation and rotating manipulation were respectively adopted in treatment group and control group (on alternate days one time, 3 times as a course of treatment). The symptoms and signs (including back pain and discomfort, lower limb pain and numbness, powerless urination and defecation, numbness in perineum, straight-leg raising degree, ability of lower extremity walking, work and live) of patients were observed after treatment.</p><p><b>RESULTS</b>All the patients were followed up from 1 to 6 months with an average of 3.2 months. After treatment, the symptoms and signs of patients have markedly improved (P < 0.01), but the lower back pain and discomfort, lower limb walking ability in treatment group were better than control group (P < 0.05). According therapeutic criteria, the effect of treatment group was better than of control group (P < 0.01). In cases with bulging type, 47 in treatment group and 45 in control group, the effect of treatment group was better than of control group (P < 0.05); in cases with hernia type, 35 in treatment group and 34 in control group, there was no significantly difference in effect between two groups (P > 0.05); in cases of free type, 25 in treatment group and 23 in control group, there was no significantly difference in effect between two groups (P > 0.05).</p><p><b>CONCLUSION</b>The global effect of lumbar backwards flexion manipulation was satisfactory than rotating manipulation for treating lumbar intervertebral disc herniation. But rotating manipulation suited to free type.</p>

Cigarette smoke extract inhibits the proliferation of alveolar epithelial cells and induces apoptosis / 生理学报

Cigarette smoke extract (CSE) contains abundant oxidants and free radicals. Oxidative stress caused by cigarette smoking results in the destruction of the alveolar cell walls and emphysema. However, there exists discrepancy about how CSE works in the process. In the present study, we observed the effect of CSE on the cell growth of type II alveolar epithelial cell-derived A549 cell line, and provided molecular understanding of this effect. The MTT assay results showed that CSE decreased the cell viability of A549 cells in a dose- and time-dependent manner, and cell cycle was arrested in G(1)/S phase. Furthermore, CSE-induced apoptosis of A549 cells was verified by Hoechst 33258 staining, electron microscopy in morphology, and the appearance of DNA fragmentation and annexin V-FITC/propidium iodide (PI) staining assay at molecular level. It was found that CSE treatment resulted in the upregulation of Fas/APO-1 receptor and activation of caspase-3. CSE also initiated accumulation of intracellular reactive oxygen species, which was detected by laser confocal microscopy. Taken together, CSE could inhibit the cell growth and induce apoptosis of A549 cells through Fas receptor pathway. Oxidative stress caused by CSE may be the radical factor leading to apoptosis as well as cell growth inhibition in alveolar epithelial cells.

Inhibition of expression of hypoxia-inducible factor-1alpha mRNA by nitric oxide in hypoxic pulmonary hypertension rats / 华中科技大学学报（医学）（英德文版）

In order to study the effect of nitric oxide (NO) on the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L-arginine (L-Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF-1alpha mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT-PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6 +/- 2.7 mmHg, 1 mmHg=0.133 kPa) was significantly lower than that in chronic hypoxic group (35.8 +/- 6.1 mmHg, t=0.2918, P<0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L-arginine group (24.4 +/- 3.8 mmHg, t=0.2563, P<0.05). ISH showed that the expression of HIF-1alpha mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076 +/- 0.0205) was markedly weaker than that in chronic hypoxic group (0.3317 +/- 0.0683, t=3.125, P<0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L-arginine group (0.1928 +/- 0.0381, t=2.844, P<0.05). RT-PCR showed that the content of HIF-1alpha mRNA in chronic hypoxic group (2.5395 +/- 0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781 +/- 0.3628) and hypoxia plus L-arginine group (1.4511 +/- 0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF-1alpha mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.

Construction of eukaryotic expression plasmid of human PRX3 and its expression in HEK-293FT cells / 华中科技大学学报（医学）（英德文版）

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.

Inhibition of expression of hypoxia-inducible factor-1alpha mRNA by nitric oxide in hypoxic pulmonary hypertension rats / 华中科技大学学报（医学）（英德文版）

In order to study the effect of nitric oxide (NO) on the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) mRNA in hypoxic pulmonary hypertension (HPH) rats, 30 healthy male Wistar rats were randomly divided into normoxic control group, chronic hypoxic group and hypoxia plus L-arginine (L-Arg) group. The animal model of HPH was developed. The mean pulmonary arterial pressure (mPAP) was measured by inserting a microcatheter into the pulmonary artery. The HIF-1alpha mRNA expression levels were detected by in situ hybridization (ISH) and semiquantitative RT-PCR. It was found that after 14 days hypoxia, the mPAP in normoxic control group (17.6 +/- 2.7 mmHg, 1 mmHg=0.133 kPa) was significantly lower than that in chronic hypoxic group (35.8 +/- 6.1 mmHg, t=0.2918, P<0.05) and mPAP in chronic hypoxic group was higher than that in hypoxia plus L-arginine group (24.4 +/- 3.8 mmHg, t=0.2563, P<0.05). ISH showed that the expression of HIF-1alpha mRNA in the intraacinar pulmonary arteriolae (IAPA) in normoxic control group (0.1076 +/- 0.0205) was markedly weaker than that in chronic hypoxic group (0.3317 +/- 0.0683, t=3.125, P<0.05) and that in chronic hypoxic group was stronger than that in hypoxia plus L-arginine group (0.1928 +/- 0.0381, t=2.844, P<0.05). RT-PCR showed that the content of HIF-1alpha mRNA in chronic hypoxic group (2.5395 +/- 0.6449) was 2.16 times and 1.75 times higher than that in normoxic control group (1.1781 +/- 0.3628) and hypoxia plus L-arginine group (1.4511 +/- 0.3981), respectively. It is concluded that NO can reduce the mPAP by the inhibition of the expression of HIF-1alpha mRNA, which may be one of the mechanisms through which NO affects the pathogenesis of HPH.

Construction of eukaryotic expression plasmid of human PRX3 and its expression in HEK-293FT cells / 华中科技大学学报（医学）（英德文版）

To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.

Melatonin in in vitro apoptosis of H22 hepatocarcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of melatonin (MLT) in in vitro apoptosis of hepatocarcinoma cells and its mechanism.</p><p><b>METHODS</b>The apoptotic cells, bcl-2 and bax were detected through immunocytochemical method (ICC) and Tolt-mediated x-duTP nick end labeling (TUNEL). Computer image analysis system was used to quantify the expression of bcl-2 and bax by detecting the absorbance value of positive products. Apoptosis index (AI) was used to quantify the number of apoptotic cells.</p><p><b>RESULTS</b>In vitro, AI increase was both concentration- and time-dependent through TUNEL. During the same duration, AI of medium dose group was higher than that of low dose and control group (P < 0.05); AI of high dose, medium dose and 5-Fu group were higher than those of low dose and control group (P < 0.01), however, there was no significant difference between the low dose and control group (P > 0.05). At the same dose, in high dose, medium dose and 5-Fu group, the change of AI showed significant difference from 24 to 36 hours (P < 0.05). The expression of bcl-2 was down-regulated as the MLT increased, and there was significant difference between the low dose and control group (P < 0.01). But, the expression of bax was up-regulated as the dose of MLT increased, showing significant difference between the high dose and control groups (P < 0.01). As time went on, the expression of bcl-2 was decreased and in every group, with the change in absorbance value of bcl-2 significantly different from 24 to 36 hours (P < 0.05), whereas that of bax remained almost unchanged. The ratio of bax/bcl-2 was increased with the increase in the concentration of MLT.</p><p><b>CONCLUSION</b>Melatonin may induce apoptosis in the hepatocarcinoma cells which is concentration- and time-dependent, in which bcl-2 and bax are involved.</p>

Changes of the actin and transforming growth factor-beta1 expression in the small airways of the rat with chronic obstructive lung disease / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the roles of actin and transforming growth factor (TGF)-beta(1) in the injury repair and the development of emphysema.</p><p><b>METHODS</b>Wistar rats were randomly divided into two groups: the smoking and infection group (group SI) and the control group (group C). The rats of group SI received smoking irritation accompanying with repeated intranasal infection. Subgroups of the experimental animals were killed in the 2nd, 4th, 8th and 16th weeks respectively. The morphological changes of lungs were compared and PaO(2), PaCO(2) as well as the right ventricular systolic pressure (RVSP) were analysed. The lung sections were stained with immunohistochemistry for actin and TGF-beta(1).</p><p><b>RESULTS</b>In comparison with animals of group C, thickening of the bronchiolar walls, narrowing of bronchiolar lumens, and area of emphysema were much severe in animals of group SI (P < 0.05). The muscularization of intra-alveolar arteries in group SI in the 16th week was apparent in comparing with that in group C (P < 0.05). PaO(2) values in group SI were significantly decreased, and RVSP values in group SI were significantly increased in the 8th and 16th week (P < 0.05). Actin expression was increased in animals of group SI in the 4th and 8th week (0.24 +/- 0.06 and 0.25 +/- 0.05) in comparing with that of group C (0.09 +/- 0.03) (P < 0.05). Animals of group SI showed a significant increase of TGF-beta(1) in lung tissue in different periods as mentioned in above (33.33 +/- 12.11, 45.71 +/- 15.12, 71.43 +/- 16.76 and 86.25 +/- 20.66 respectively).</p><p><b>CONCLUSIONS</b>The increased expression of actin and TGF-beta(1) protein in small airways induced by smoking irritation and Klebsiella Pneumoniae may interfere with the repair response, and contributes to the development of emphysema.</p>

Expression of hypoxia-inducible factor-1 alpha and endothelin-1 gene in hypoxic pulmonary hypertension / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of hypoxia-inducible factor-1 alpha (HIF-1alpha) and endothelin-1 (ET-1) gene in hypoxic pulmonary hypertension (HPH).</p><p><b>METHODS</b>The animal model of HPH was replicated. The elastic fiber staining was applied to show the intraacinar pulmonary artery (IAPA). Radioimmunoassay (RIA) and in situ hybridization (ISH) were used for detection of HIF-1a and. ET-1.</p><p><b>RESULTS</b>ISH showed that HIF-1alpha mRNA was expressed in the IAPA of all hypoxic rat. The expression was stronger in the H14 d (0.256 9 +/- 0.046 8) and H28 d (0.225 8 +/- 0.045 3) groups than in the H5 d (0.1455 +/- 0.072 2) and control (0.110 9 +/- 0.022 4) groups (P < 0.05), the expression of ET-1 mRNA in the H14 d (0.412 2 +/- 0.078 3) and H28 d (0.368 4 +/- 0.072 9) groups was also stronger than that in the H5 d (0.201 7 +/- 0.034 9) and control (0.185 5 +/- 0.036 1) groups (P < 0.05). The amount of ET-1 in pulmonary arteial blood in the H14 d [(158.78 +/- 25.14) pg/ml] and H28 d [(142.93 +/- 23.38) pg/ml] groups was significantly higher than that in the H5 d [(79.68 +/- 12.54) pg/ml] and control [(65.37 +/- 10.82) pg/ml] groups (P < 0.05). The mean pulmonary arterial pressure (mPAP) in the H14 d [(34.0 +/- 5.8) mm Hg] and H 28 d [(29.0 +/- 4.7) mm Hg] groups was markedly higher than that in the H5 d [(19.0 +/- 3.5) mm Hg] and control [(17.0 +/- 2.8) mm Hg] groups (P < 0.05). A positive rank correlation existed between the mPAP and the amount of ET-1 (rs = 0.747, P < 0.05).</p><p><b>CONCLUSIONS</b>Expression of HIF-1alpha and ET-1 mRNA in IAPA increase under long-term hypoxic condition and both show consistent expression, indicating that the expression of HIF-1a and ET-1 gene contribute to pathogenesis of HPH.</p>

Expression of phosphatidylinositol 3-kinase during the proliferation of the hypoxic rat pulmonary arterial smooth muscle cells / 中国病理生理杂志

AIM: To investigate the expression of phosphatidylinositol 3-kinase (PI-3K) during the proliferation of the hypoxic pulmonary arterial smooth muscle cells(PASMC). METHODS: The cultured PASMC was divided into two groups: serum-free medium (SFM) group and 48 hours hypoxia (H48h) group. Immunohistochmistry(IHC) and Flowcytometer(FCM) were used to detect the cultured PASMC. RESULTS: The positive expression of PI-3K p110 existed in both cultured PASMC groups. The staining intensity in H48h group (0 1891?0 0301) was significantly stronger than that in SFM group (0 1025?0 0164, P

Effects of cigarrete smoke extract and lipopolysaccharid e on the expression of transforming growth factor-?_ 1 mRNA and protein in human embryonic lung fibroblasts / 中国病理生理杂志

AIM: To examine the effects of cigarette smo ke extract (CSE) and lipo polysaccharide (LPS) on the production of transforming growth factor-? 1 (TGF- ? 1 ) mRNA and protein in cultured human embryonic lung fibroblasts (HELF). METHODS: The cultured HELF were incubated with CSE, LPS or CSE in com bination with LP S for 24 hours at 37℃, respectively. The total RNA was extracted from the cells . The expression levels of TGF-? 1 mRNA were evaluated by reverse transcription - p olymerase chain reaction (RT-PCR). The mRNA levels of TGF-? 1 were corrected by GAPDH transcripts, and TGF-? 1 protein levels were determined by immunocytoch emistry. RESULTS: CSE stimulated the TGF-? 1 mRNA expression i n HELF at lower concentr ati ons (1∶50 and 1∶25)( P 0.05). LPS enhanced TGF-? 1 mRNA levels in HELF at a ll three doses (0.1 mg/L, 1 mg/L, and 10 mg/L)( P

Changes of nuclear factor-?B and inducible nitric oxide synthase in hypoxic pulmonary hypertension / 中国病理生理杂志

AIM:To observe the changes of nuclear factor-?B (NF-?B) activity and inducible nitric oxide synthase (iNOS) expression in hypoxic pulmonary hypertension(HPH). METHODS:The rat model of HPH was used. The NF-?B activity and iNOS expression in lung tissue were determined by immunohistochemistry(IHC), in situ hybridization (ISH), RT-PCR and Western blot.RESULTS:ISH showed that iNOS mRNA expression in intraacinar pulmonary arteriole (IAPA) in H28 d group(hypoxic treatment for 28 days) was stronger than that in normal group, H5d group and H14 d group. RT-PCR showed that the iNOS mRNA in H28 group was 2.1 times, 1.9 times and 1.8 times higher than that in normal group, H5d group and H14 d group, respectively. The nucleic anti-NF-?B stain was observed in H28 d group, which was significantly stronger, but the I-?B amount was 2.8 times, 2.7 times and 2.5 times lower than that in normal group, H5d group and H14 d group, respectively. CONCLUSION: The activity of NF-?B was correlated with the hypoxic pulmonary vessel structural remodeling and iNOS expression.