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Abstract Background Tandem mass spectrometry (MS/MS) analysis is a powerful tool for newborn screening, and many rare inborn errors of metabolism are currently screened using MS/MS. However, the sensitivity of MS/MS screening for several inborn errors, including citrin deficiency (screened by citrulline level) and carnitine uptake defect (CUD, screened by free carnitine level), is not satisfactory. This study was conducted to determine whether a second-tier molecular test could improve the sensitivity of citrin deficiency and CUD detection without increasing the false-positive rate. Methods Three mutations in the SLC25A13 gene (for citrin deficiency) and one mutation in the SLC22A5 gene (for CUD) were analyzed in newborns who demonstrated an inconclusive primary screening result (with levels between the screening and diagnostic cutoffs). Results The results revealed that 314 of 46 699 newborns received a second-tier test for citrin deficiency, and two patients were identified; 206 of 30 237 newborns received a second-tier testing for CUD, and one patient was identified. No patients were identified using the diagnostic cutoffs. Although the incidences for citrin deficiency (1:23 350) and CUD (1:30 000) detected by screening are still lower than the incidences calculated from the mutation carrier rates, the second-tier molecular test increases the sensitivity of newborn screening for citrin deficiency and CUD without increasing the false-positive rate. Conclusions Utilizing a molecular second-tier test for citrin deficiency and carnitine transporter deficiency is feasible.

This work aimed to clarify the potential regulating effects of Qufeng Xuanfei formula (QFXF) on airway neurogenic inflammation and its underlying target signal pathway. Guinea pig model of airway hyperergy (AHR) was used. The relative susceptibility of major proteins to airway neurogenic inflammation was assessed using Western blot immunoassay followed by being separated by SDS-PAGE. Compared to the model group, QFXF of all concentrations effectively depressed the capsaicin enhanced cough in guinea pigs and the peak values of airway resistance significantly decreased. The results illustrated that QFXF alleviated cough symptom in guinea pigs and reduced airway neurogenic inflammation when compared to AHR model group. Airway inflammation and damage, as well as the levels of NGF, SP and c-Fos in QFXF decreased the most in the high-dose group. The mechanism of antitussive activity may be associated with reducing airway inflammation. QFXF displayed effect on chronic cough through reducing the levels of neuropeptides, attenuating airway inflammation and promoting recovery from disease to decrease the airway neuro sensitivity, suggesting that the potential mechanism may be related to Ras/ERK/c-Fos pathway.

Objective: To investigate the protective effects of paeoniflorin against PM2.5-induced damage in BEAS-2B cells and explore the possible mechanism., Methods: With a factorial design, this study was performed to observe the protective effects of different doses of paeoniflorin against PM2.5-induced BEAS-2B cell growth inhibition and the effects of paeoniflorin on the contents of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS) in the cell cultures., Results: Exposure to increased PM2.5 concentrations caused significant decrease in the cell survival rate (P<0.05) with a clear dose-response relationship (r=-0.759, P<0.05). Treatment of the cells with paeoniflorin significantly attenuated PM2.5-induced inhibition of BEAS-2B cell survival (P<0.05), but the effect of paeoniflorin was not dose-dependent (P>0.05). PM2.5 exposure also significantly increased the contents of MDA and intracellular ROS (P<0.05), and paeoniflorin obviously antagonized these effects of PM2.5., Conclusion: Paeoniflorin can protect BEAS-2B cells from PM2.5-induced growth inhibition, and the mechanism might be related to the anti-oxidant effects of paeoniflorin.

Objective: To investigate the effect of low-selenium diet on the liver and kidneys of rats and explore the role of macrophage polarization into M1 and M2 phenotypes in liver and kidney injuries., Methods: Twenty-four rats (12 female and 12 male) were randomly divided into control group and low-selenium group and fed with normal chow (dietary selenium of 0.18 mg/kg) and low-selenium diet (dietary selenium of 0.02 mg/kg) for 109 days. After the feeding, the rats were sacrificed for HE staining to observe liver and kidney pathologies, and immunohistochemistry was performed for analyzing CCR7, CD206, CD163-positive cell numbers in the liver and kidneys., Results: The rats in low-selenium group showed severer fibrosis in the liver and kidney than the control group. In either male or female rats in low-selenium group, CCR7 and CD206 expressions in the liver were comparable with those in control group, but CD163 expression was lower than that in the control group (P<0.05 for both female and male rats). In the kidney, the proximal tubule showed a slightly higher while the distal tubule showed a slightly lower CCR7 expression in low selenium group than in the control group (P>0.05). In low-selenium group, a significantly lower CD163 expression in the distal tubule and a significantly higher CD206 expression in the proximal tubule were noted as compared with the control group (P<0.05 in both female and male rats). Compared with the control rats, the male rats in low-selenium group, but not the female rats, showed a significantly lower CD163 expression in the proximal tubule of the kidney (P<0.05); the female but not the male rats in low-selenium group show a higher CD206 expression in the distal tubule (P<0.05)., Conclusion: Low-selenium diet can cause liver and kidney fibrosis in rats and may inhibit macrophage activation into the M2 phenotype.

Background: Pathogenesis and effective therapeutics of chronic pancreatic inflammation and fibrosis remain uncertain., Purpose: To investigate the effects of sulfasalazine (SF) on pancreatic inflammation and fibrogenesis., Methods: Chronic pancreatic injury in rats was induced by diethyldithiocarbamate (DDC) and interfered by SF through intraperitoneal injection. The rats were divided into five groups: group N, normal control group, rats were treated with dilated water only; group DS1, rats received SF (10 mg/kg) 2 hours before DDC treatment; group DS2, rats were treated with DDC and then SF (100 mg/kg, twice a week); group DS3, rats were treated with DDC, then SF (100 mg/kg, thrice a week); and group DDC, rats were treated with DDC only. Pancreatic inflammation and fibrosis were determined by hematoxylin and eosin staining and Sirius red staining. The genes and proteins related to NF-κB pathway and fibrogenesis including NF-κB/p65, TNF-α, ICAM-1, α-SMA, and Con 1 were detected by immunohistochemical staining, reverse transcription polymerase chain reaction, and Western blotting., Results: Rats in the DDC and DS1 groups showed the highest histological scores after DDC treatment, but the scores of DS2 and DS3 groups decreased significantly when compared with the DDC group. Sirius red staining showed collagen formation clearly in DDC and DS1 rats rather than in DS2 and DS3 rats. NF-κB/p65, ICAM-1, and α-SMA were strongly expressed in DDC and DS1 rats, while DS2 and DS3 rats showed mild to moderate expression by immunohistochemistry. Reverse transcription polymerase chain reaction showed increased levels of NF-κB/p65, ICAM-1, TNF-α, α-SMA, and Con 1 mRNA in DDC and DS1 rats in comparison to normal controls. The mRNA levels of these molecules in DS2 and DS3 rats were significantly lower than those in DS1 and DDC rats. Western blotting demonstrated that the NF-κB/p65, ICAM-1, and α-SMA expressions in pancreatic tissues of the rats of the DDC group were more clear than those of the normal control, DS2, and DS3 rats., Conclusion: SF inhibits pancreatic inflammation and fibrogenesis via NF-κB signaling pathway.

Objective: To study the effects and role of central endocannabinoid system in the mechanism of visceral hyposensitivity induced by rapid eye movement (REM) sleep deprivation., Methods: Twenty-four SD rat were divided randomly in to 3 groups: cage-yoked group (YC Group, experimental control group), REM sleep deprivation group (SD Group) exposed to REM sleep deprivation by means of flower pot technique lasting for 48 hours, and Rim Group, receiving rimonabant, a cannabinoid antagonist, after REM sleep deprivation. 48 hours after the sleep deprivation abdominal electromyogram in response to colorectal distension (CRD) was recorded to asses the visceral sensitivity. Then the rats were killed with their central nervous system taken out. RT-PCR and Western blotting were used to detect the RNA and protein expression of cannabinoid receptor CB1, fatty acid amide hydrolase (FAAH), and monoacylglycerol lipase (MGL) in the thalamus, brain stem, and spinal cord., Results: (1) Under the pressures of 40, 60, and 80 mm Hg, the abdominal electrical activity frequencies of external oblique muscle responding to CRD in SD Group were (220 +/- 94), (313 +/- 162), and (493 +/- 279) times respectively, all significantly lower than those in YC Group [(506 +/- 223), (1053 +/- 548), and (1632 +/- 249) times respectively, all P < 0.05], and those of Rim Group were (668 +/- 257), (1144 +/- 93), and (1653 +/- 153) times respectively, all significantly higher than those of SD Group (all P < 0.05), but not significantly different from those of YC Group. (2) The RNA and protein expression levels of CB1 receptor in the thalamus, brain stem, and spinal cord of SD Group were all significantly higher than those of YC Group (all P < 0.05), while the RNA and protein expression levels of FAAH and MGL in the thalamus and spinal cord of SD Group were all significantly lower than those of YC Group (all P < 0.05)., Conclusion: The visceral hyposensitivity induced by REM sleep deprivation may be associated with the increase of expression of CNS endocannabinoid receptor and decrease of its metabolism.