Indole Propionic Acid Disturbs the Normal Function of Tryptophanyl-tRNA Synthetase in Mycobacterium tuberculosis.

Tuberculosis (TB) is the leading infectious disease caused by Mycobacterium tuberculosis and the second-most contagious killer after COVID-19. The emergence of drug-resistant TB has caused a great need to identify and develop new anti-TB drugs with novel targets. Indole propionic acid (IPA), a structural analog of tryptophan (Trp), is active against M. tuberculosis in vitro and in vivo. It has been verified that IPA exerts its antimicrobial effect by mimicking Trp as an allosteric inhibitor of TrpE, which is the first enzyme in the Trp synthesis pathway of M. tuberculosis. However, other Trp structural analogs, such as indolmycin, also target tryptophanyl-tRNA synthetase (TrpRS), which has two functions in bacteria: synthesis of tryptophanyl-AMP by catalyzing ATP + Trp and producing Trp-tRNATrp by transferring Trp to tRNATrp. So, we speculate that IPA may also target TrpRS. In this study, we found that IPA can dock into the Trp binding pocket of M. tuberculosis TrpRS (TrpRSMtb), which was further confirmed by isothermal titration calorimetry (ITC) assay. The biochemical analysis proved that TrpRS can catalyze the reaction between IPA and ATP to generate pyrophosphate (PPi) without Trp as a substrate. Overexpression of wild-type trpS in M. tuberculosis increased the MIC of IPA to 32-fold, and knock-down trpS in Mycolicibacterium smegmatis made it more sensitive to IPA. The supplementation of Trp in the medium abrogated the inhibition of M. tuberculosis by IPA. We demonstrated that IPA can interfere with the function of TrpRS by mimicking Trp, thereby impeding protein synthesis and exerting its anti-TB effect.

Pregnancy characteristics and adverse outcomes in offspring of women with epilepsy: a prospective registry study from Mainland China.

Objective: This study aimed to explore the influencing factors of adverse outcomes in the offspring of women with epilepsy (WWE) and to analyze the changes brought about by the epilepsy knowledge popularization campaign in China (EKPCIC). Methods: This nested case-control study focused on WWE and their offspring from a female epilepsy cohort in mainland China. From January 2009 to August 2022, WWE was prospectively enrolled in 32 study centers. This study aimed to observe the health outcomes of their offspring within 1 year of age. The main outcome measure assessed the health status of the offspring within their first year of age. We aimed to analyze the effects of seizures, anti-seizure medicines (ASMs), and a lack of folic acid supplementation on adverse outcomes in the offspring of WWE and to explore the changes in perinatal management and adverse outcomes of the offspring after dissemination of the EKPCIC in 2015. Additionally, subgroup analyses were conducted to compare seizure control during pregnancy between the valproate and non-valproate groups. Results: In total, 781 pregnancies in 695 WWE were included, of which 186 (23.69%) had adverse outcomes. The National Hospital Epilepsy Severity Scale score, number of seizures, status epilepticus, ASM type, and valproate and folic acid doses were associated with a high risk of adverse outcomes. After the EKPCIC, the use of ASMs (P = 0.013) and folic acid (P < 0.001), the seizure-free rate during pregnancy (P = 0.013), and the breastfeeding rate (P < 0.001) increased, whereas the incidence of complications during pregnancy decreased (P = 0.013). However, there was no significant difference in the incidence of adverse outcomes between the analyzed offspring pre-/post-EKPCIC. Additionally, there was no association between the frequency of seizures at different time points during pregnancy and the use of valproate (F = 1.514, P = 0.221). Conclusion: Possible factors influencing adverse outcomes in the offspring of WWE include seizures, type and number of ASM usage, and a lack of folic acid supplementation. Although the management of WWE during pregnancy is now more standardized, further efforts are needed to reduce adverse outcomes in offspring.

Li-Dan-He-Ji Improves Infantile Cholestasis Hepatopathy Through Inhibiting Calcium-Sensing Receptor-Mediated Hepatocyte Apoptosis.

Infantile cholestatic hepatopathy (ICH) is a clinical syndrome characterized by the accumulation of cytotoxic bile acids in infancy, leading to serious liver cirrhosis or liver failure. The aetiology of ICH is complicated and some of them is unknown. Regardless of the aetiology, the finial pathology of ICH is hepatocyte apoptosis caused by severe and persistent cholestasis. It is already known that activation of calcium-sensing receptor (CaSR) could lead to the apoptosis of cardiomyocytes. However, the mechanism by CaSR-mediated cholestasis-related hepatocyte apoptosis is not fully understood. Li-Dan-He-Ji (LDHJ), a Traditional Chinese Medicine prescription, was developed to treat ICH. Another aim of this study was to investigate the possible mechanisms of LDHJ in cholestasis-related hepatocyte apoptosis. Using the primary hepatocytes, we first investigated the molecular mechanism of CaSR-mediated hepatocyte apoptosis in cholestasis. Then we prepared LDHJ granules and used ultra-high-performance liquid chromatography to identify the predominant drugs; confirmed the stability of the main substances; and for cell experiments screened forsythoside-A, emodin and chlorogenic acid as the three active substances of LDHJ granules. In the young rats with ANIT-induced intrahepatic cholestasis and the primary hepatocytes with TCDC-induced cholestasis-related hepatocyte apoptosis, the levels of liver injury and cholestasis-related biomarkers, calcium-sensing receptor (CaSR), hepatocyte apoptosis, Bax/Bcl-2 ratio, Cytochrome-C, caspase-3, phosphorylated-c-Jun NH2-terminal kinase (p-JNK)/JNK, and p-P38/P38 were all increased, while the levels of p-extracellular signal-regulated kinase (p-ERK)/ERK were decreased. However, LDHJ granules and its three active substances effectively reversed these changes. Furthermore, the three active substances reduced the increases in the intracellular calcium concentration ([Ca2+]i) and ROS levels and attenuated the dissipation of the mitochondria membrane potential in the TCDC-induced primary hepatocytes. The opposite results were obtained from the TCDC-induced primary hepatocytes treated with an agonist of CaSR (GdCl3) plus forsythoside-A, emodin or chlorogenic acid. Based on the results from in vivo and in vitro studies, LDHJ functions as an antagonist of CaSR to regulate hepatocyte apoptosis in cholestasis through the mitochondrial pathway and mitogen-activated protein kinase pathway.

[Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells].

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using palmitic acid( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-ß1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-ß1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-ß1.

[Effect of Psoralea corylifolia in treating fatty liver disease in juvenal mouse by inhibiting hepatic NF-κB activation].

To investigate the mechanism and effect of Psoralea corylifolia(PC) in the treatment of NAFLD in juvenal mice. The NAFLD model in juvenal mice was established by feeding high-fat diet. Then PC herbal granules (at low and high dose) were administered for 5 weeks. Blood glucose (FBG, PG-1 h/2 h), blood lipid (TC, TG, HDL-C, LDL-C), fasting insulin, liver function (ALT, AST) were examined. HOMA-IR was calculated. Hepatic histological changes were observed. The content of TG, inflammatory factor (TNF-α, IL-8) and protein expressions of CD44, NF-κB p65, p-NF-κB p65 in hepatic tissues were determined. The ratio of p-NF-κB p65 to NF-κB p65 (p-p65/p65) was calculated. The result showed that compared with the model group, both PC treatment groups showed reduction in hepatic steatosis, inflammatory cell infiltration and fibroplasia in portal area. HOMA-IR, ALT, AST, FBG, PG-2 h, TC, TG, LDL-C concentrations and hepatic TG content were also significantly decreased, with the reduction of TNF-α, IL-8 contents, CD44 expression and p-p65/p65 ratio in hepatic tissues (P<0.01). High-dose PC group had a better effect than low-dose group (P<0.01, P<0.05). In conclusion, PC is effective in treating hepatic injury, glucolipid metabolism disturbances and fibrosis in juvenal NAFLD mice. The mechanism may be related to inhibition of inflammation and down-regulation of the activation of hepatic NF-κB.

[Protective effect of emodin pretreatment in young rats with intrahepatic cholestasis].

OBJECTIVE: To investigate the protective effect of emodin in young rats with intrahepatic cholestasis. METHODS: A total of 120 young Sprague-Dawley rats were randomly divided into control, model, and high-, medium-, and low-dose emodin groups, with 24 rats in each group. The rats in the control and model groups were given sodium carboxymethyl cellulose solution by gavage, while the other groups were given different doses of emodin solution by gavage. On the 5th day of experiment, alpha-naphthylisothiocyanate (ANIT, 50 mg/kg) was applied by gavage to establish the model of intrahepatic cholestasis in all groups except the control group. At 24, 48, and 72 hours after gavage, 8 rats in each group were sacrificed. Colorimetry was used to measure the serum levels of total bilirubin (TBIL), direct bilirubin (DBIL), total bile acid (TBA), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (GGT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in each group, and hematoxylin-eosin staining was applied to observe the morphological changes of the liver under a light microscope at different time points. RESULTS: Compared with the control group, the model group had significantly increased serum levels of TBIL, DBIL, TBA, ALP, GGT, ALT, and AST at the 24-hour, 48-hour, and 72-hour time points (P<0.01). In the model group, the serum levels of TBIL, DBIL, TBA, ALT, and AST showed varying degrees of increase at 48 hours after establishment of model, compared with the values at 24 and 72 hours (P<0.05). At 24, 48, and 72 hours, the high-, medium-, and low-dose emodin groups had varying degrees of reductions in the serum levels of TBIL and TBA compared with the model group (P<0.05); the high- and low-dose emodin groups had significantly increased serum levels of TBA compared with the medium-dose emodin group (P<0.05). The model group had the most severe pathological changes at 48 hours. Compared with the model group, the high-, medium-, and low-dose emodin groups showed certain improvement in pathological changes of the liver at each time point, and the medium-dose emodin group had better improvement compared with the high- and low-dose emodin groups. CONCLUSIONS: Emodin can effectively improve ANIT-induced intrahepatic cholestasis in young rats, and medium-dose emodin shows the best effect.

Preventative effect of massage on gastric volvulus in infants with gastroesophageal reflux-induced pneumonia.

OBJECTIVE: To study the preventative effects of massage on gastric volvulus (GV) in infants with gastroesophageal reflux (GER)-induced pneumonia. METHODS: One-hundred and eighty GV with GER-induced pneumonia inpatients were divided randomly into four groups: basic treatment 1 (n = 60), basic treatment 2 (n = 30), massage treatment 1 (n = 60) and massage treatment 2 (n = 30). Clinical examinations selected between groups 1 and 2 were different. Radiography of the upper gastrointestinal tract using iodine-containing contrast was assessed in group 1 before and after treatment, whereas 24-h pH monitoring of the distal esophagus was assessed in group 2 before and after treatment. Symptom scores and chest radiography were assessed in all groups upon hospital admission and after procedures. Clinical effects were estimated after procedures in all groups. The prevalence of severe pneumonia among the four groups was compared. RESULTS: Massage treatment groups showed a significantly higher percentage of cure and total effect (P < 0.05, P < 0.01) and a lower prevalence of recurrence (but with no statistic difference, P > 0.05) than basic treatment groups. Furthermore, massage treatment groups had remarkably lower scores for symptoms and signs (P < 0.05, P < 0.01), especially for choking on milk, than basic treatment groups. There was significant attenuation of chest inflammation (P < 0.05, P < 0.01), GV (P < 0.05, P < 0.01) and GER (P < 0.05, P < 0.01) in massage treatment groups compared with those in basic treatment groups. Finally, massage treatment groups demonstrated a lower prevalence of severe pneumonia than basic treatment groups (P < 0.05). CONCLUSION: Massage treatment can prevent GV with GER-induced pneumonia in infants by timely correction of stomach rotation and subsequent attenuation of GER.

[Clinical observation on chiropractics combined with acupuncture at Sifeng (EX-UE 10) for treatment of infantile anorexia].

OBJECTIVE: To search for an effective therapy for infantile anorexia. METHODS: Sixty-two cases of infantile anorexia were randomly divided into a treatment group and a control group, 31 cases in each group. The treatment group were treated with chiropractics plus acupuncture at Sifeng (EX-UE 10), and the control group with oral administration of zinc gluconate granules made by the pharmaceutical factory of the hospital, for 2 weeks. RESULTS: The effective rate was 93.5% in the treatment group, which was better than 74.2% in the control group (P < 0.05); there were significant differences in blood zinc content and urinary amylase activity in the two groups before and after treatment (P < 0.01), with more significant increase in the treatment group as compared with the control group (P < 0.05). CONCLUSION: Chiropractics combined with acupuncture at Sifeng (EX-UE 10) has a better therapeutic effect on infantile anorexia.