Discrimination of Baphicacanthis Cusiae Rhizoma et radix and its adulterant species and establishment of an assay method for quality control.

BACKGROUND: Baphicacanthis Cusiae Rhizoma et Radix, commonly known as Nan-Ban-Lan-Gen (NBLG), is an essential traditional Chinese medicine that possesses diverse bioactivities, particularly noteworthy for its antiviral properties. Although NBLG has been listed in the Chinese Pharmacopoeia as an independent Chinese medicine, the establishment of a comprehensive quality standard for NBLG remains elusive. The absence of assay for marker compound in its quality standards has led to the lack of corresponding quality control measures for NBLG-containing preparations, and its discrimination from adulterant species in the market which thereby can significantly impact the efficacy and safety of NBLG-containing products. METHODS: Ultra-high performance liquid chromatography (UHPLC) coupled with quadrupole-time-of-flight mass spectrometry (Q-TOF-MS) was employed for comprehensive profiling of the chemical constituents of NBLG, the stem of Baphicacanthus cusia (Nees) Bremek (NBLJ), and the roots of Isatis indigotica Fort. (Bei-Ban-Lan-Gen, BBLG). Additionally, multivariate analysis was conducted to compare the chemical components of NBLG with those of NBLJ and BBLG. Furthermore, we established an optimized and validated HPLC method to obtain the fingerprint of NBLG and quantify the content of 2-benzoxazolinone and acteoside in the samples. RESULTS: A total of 73 compounds belonging to six classes were assigned in NBLG, with alkaloids being the most abundant and diverse species. High compositional similarities with significant differences in content were observed between NBLG and NBLJ. Moreover, the chemical profile of BBLG markedly differed from that of NBLG. An informative high performance liquid chromatography (HPLC) fingerprint of NBLG comprising seven characteristic peaks that can be used for quality assessment was established. Notably, we propose a quality control standard for NBLG, stipulating that the limit of content in dry weight for both 2-benzoxazolinone and acteoside should not be less than 0.010%. CONCLUSION: This study provides the most comprehensive chemical information to date on NBLG, offering valuable insights into its authentication and quality control. Our findings highlight the importance of comprehensive chemical profiling to differentiate potential substitutions and adulterations of herbal medicines, particularly when the original source is scarce or unavailable. These results can aid in the development of quality control measures for NBLG-containing preparations, ensuring their safety and efficacy.

Peroxiredoxin 1 transfer during mating protects eupyrene sperm against oxdative stress in Grapholita molesta.

BACKGROUND: Each Grapholita molesta female only copulates once during its lifetime and thus must maintain the viability of stored eupyrene sperm for male reproductive success. The male ejaculate comprises abundant accessory gland proteins produced by the male accessory gland (AG), and many of which are major effectors for sperm storage and maintenance. RESULTS: Here, we reported that an antioxidant protein, peroxiredoxin 1 (GmolPrx1), secreted by the male AG, is essential for protecting eupyrene sperm from oxidative stress and maintaining their quality during storage in the female bursa copulatrix (BC). Our data showed that GmolPrx1 is highly expressed in the AG of sexually mature males. The GmolPrx1 protein is localized to the cytoplasm of AG cells and delivered to the female BC during mating. Knockdown of GmolPrx1 strongly decreased the fertility of mated females. Additionally, we evaluated oxidative status in the spermatophore of females and found that the content of hydrogen peroxide increased significantly after mating with GmolPrx1 knockdown males. Finally, the quality assessment of eupyrene sperm demonstrated that the plasma membrane integrity, acrosome integrity, and DNA integrity were all severely impaired in the spermatophore of females after mating with GmolPrx1 knockdown males, which may contribute to the fertility decline in males. CONCLUSION: Our current data demonstrated that activities of eupyrene sperm stored in females can be significantly impaired by enhanced oxidative stress through knocking down of GmolPrx1 in males. Our finding thus may further lay new foundations for the control of G. molesta through suppressing their populations by manipulating male reproductive genes. © 2023 Society of Chemical Industry.

Intrahepatic Cholestasis of Pregnancy Increases Inflammatory Susceptibility in Neonatal Offspring by Modulating Gut Microbiota.

Intrahepatic cholestasis of pregnancy (ICP) is a liver disease of pregnancy that is characterized by increased bile acid levels in maternal serum. Studies have shown that cholestatic pregnancy can result in long-term metabolic disturbances in the offspring. However, how ICP shapes the offspring's immunity and predisposition to inflammatory disorders at an early stage is unknown. In this study, we investigated the effect of maternal cholestasis on neonatal offspring metabolism and immune function. We compared 71 neonates with ICP mothers and 63 neonates with healthy mothers and found that the incidence of jaundice and infection was significantly higher in ICP offspring. Maternal serum total bile acid level was associated with blood cell counts in full-term ICP offspring. In animal experiments, a compensatory activation of hepatic and ileal farnesoid X receptor (FXR) and altered gut microbiota in the first week were found in ICP offspring. We also investigated lipopolysaccharide (LPS)-induced inflammatory responses in neonatal rats and found that ICP offspring were more susceptible to inflammation. To understand the correlation between congenital abnormal FXR activation and tissue immunity dysregulation, we assessed the effects of the FXR agonist GW4064 and FXR antagonist E/Z-GS in ICP offspring after LPS exposure. The expression of several pro-inflammatory cytokines significantly decreased after treatment with E/Z-GS but increased after treatment with GW4064. Treatment with the probiotic Lactobacillus rhamnosus LRX01 that inhibits FXR expression in the ileum reduced susceptibility to LPS exposure in ICP offspring. The current study indicated that cholestatic pregnancy may increase the susceptibility of the offspring to inflammation by altering bile acid metabolism and gut microbiota at an early stage. We suggest that supplementation with Lactobacillus rhamnosus LRX01, which inhibits FXR expression in the ileum, may improve intestinal immunity in ICP offspring.

SNX10 gene mutation in infantile malignant osteopetrosis: A case report and literature review / 中南大学学报(医学版)

A case of SNX10 gene mutation in a patient with infantile malignant osteopetrosis (IMO) was admitted to Department of Pediatrics, Third Xiangya Hospital, Central South University. The patient had the symptom of anemia, hepatosplenomegaly and growth retardation. The X-ray examination suggested extensive increase of bone density throughout the body, which was clinically diagnosed as IMO. The homozygous mutation of SNX10 gene c.61C>T was found via gene sequencing. We reviewed the relevant literatures and found that anemia, visual and hearing impairment, hepatosplenomegaly are the main clinical symptoms of IMO, SNX10 gene mutation is a rare cause of IMO, and hematopoietic stem cell transplantation is an effective treatment.

New phenotype of severe neonatal episodic laryngospasm due to a missense mutation in SCN4A: A case report and literature review / 中南大学学报(医学版)

Severe neonatal episodic laryngospasm (SNEL) is an ion channel disease characterized by recurrent life-threatening myotonia of respiratory muscle due to mutations in the voltage-gated sodium channel genes. Here we reported a newborn manifested as paroxysmal cyanosis and limb myotonia after birth. The neonate also developed muscle hypertrophy and stunted growth during the follow-up. Whole exome sequencing confirmed c.2395G>A, p. Ala799Thr heterozygous mutation of SCN4A. Carbamazepine was found to be effective on treating the disease. This case expands our understanding of the phenotype resulting from SCN4A mutations. By summarizing the characteristics of reported 16 cases in SNEL, we found they were mainly in the p.G1306E mutation. The common symptoms were upper airway muscle stiffness and feeding difficulties during neonates. When grow up, most patients have different degrees of recurrent attacks of myotonia and progressed muscle hypertrophy. Some of them have athlete-like special faces but all showed myotonic discharge in eletromyogram.

[Abnormal processing characteristics to basic emotional faces in the early phase in children with autism spectrum disorder].

OBJECTIVE: To explore the recognition ability and abnormal processing characteristics to basic emotional faces in the early phase in children with autism spectrum disorders (ASD). METHODS: Photos of Chinese static faces with four basic emotions (fearful, happy, angry and sad) were used as stimulus. Twenty-five ASD children and twenty-two age- and gender-matched typical developed children (normal controls) were asked to match the emotional faces with words. Event-related potential (ERP) data were recorded concurrently. RESULTS: N170 latencies for total emotion and fearful face in the left temporal region were faster than in the right one in normal controls (P<0.05), but the results were not noted in ASD children. Further, N170 latencies in the left temporal region of ASD children were slower than normal controls for total emotion, fearful and happy faces (P<0.05), and their N170 latencies in the right temporal region were prone to slower than normal controls for angry and fearful faces. CONCLUSIONS: The holistic perception speed of emotional faces in the early cognitive processing phase in ASD children is slower than normal controls. The lateralized response in the early phase of recognizing emotional faces may be aberrant in children with ASD.

Abnormal processing characteristics to basic emotional faces in the early phase in children with autism spectrum disorder / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the recognition ability and abnormal processing characteristics to basic emotional faces in the early phase in children with autism spectrum disorders (ASD).</p><p><b>METHODS</b>Photos of Chinese static faces with four basic emotions (fearful, happy, angry and sad) were used as stimulus. Twenty-five ASD children and twenty-two age- and gender-matched typical developed children (normal controls) were asked to match the emotional faces with words. Event-related potential (ERP) data were recorded concurrently.</p><p><b>RESULTS</b>N170 latencies for total emotion and fearful face in the left temporal region were faster than in the right one in normal controls (P<0.05), but the results were not noted in ASD children. Further, N170 latencies in the left temporal region of ASD children were slower than normal controls for total emotion, fearful and happy faces (P<0.05), and their N170 latencies in the right temporal region were prone to slower than normal controls for angry and fearful faces.</p><p><b>CONCLUSIONS</b>The holistic perception speed of emotional faces in the early cognitive processing phase in ASD children is slower than normal controls. The lateralized response in the early phase of recognizing emotional faces may be aberrant in children with ASD.</p>

The amplitude integrated electroencephalogram feature of term newborns with different blood glucose level / 中国新生儿科杂志

Objective To study the characteristics of amplitude integrated electroencephalogram (aEEG) in full-term newborns with different blood glucose levels,so as to provide clinical evidence for assessing brain function after hypoglycemia.Method Full-term neonates admitted to the neonatal ward of the Third Xiangya Hospital of Central South University from June 2014 to May 2016 with the initial diagnosis of hypoglycemia were enrolled to hypoglycemia group.According to the lowest level of blood glucose,infants were assigned to three subgroups,severe hypoglycemia group (＜ 1.1 mmol/L),moderate hypoglycemia group (1.1 ～ ＜2.2 mmol/L),and mild hypoglycemia group (2.2 ～ ＜2.8 mmol/L).Time matched asymptomatic term infants,who were admitted to the neonatal ward due to maternal high risks and with normal blood glucose after birth,were enrolled to control group.A 4 h continuous aEEG monitoring was completed for each infant in hypoglycemia group within 12 h after the blood glucose level stabilized.The newborns in control group were given aEEG examination 72 ～ 120 h after birth,the duration of monitoring was also 4 h.The aEEG scoring was completed and compared by rank sum test.Result A total of 83 neonates were enrolled in hypoglycemia group,including 11 with severe hypoglycemia,32 with moderate hypoglycemia,and 40 with mild hypoglycemia.Another 26 neonates with normal blood glucose level were enrolled in control group.The incidence of pregnancy-induced maternal blood glucose elevation was statistically significant among each group (P ＜ 0.05).The duration of neonatal hypoglycemia in severe hypoglycemia group was longer than that in moderate hypoglycemia group and mild hypoglycemia group [38.3 (20.7,50.4) h vs.20.4(15.3,22.6) h,13.7 (7.8,19.4) h] (P＜ 0.05).The range of glucose level in severe hypoglycemia group was larger than that in mild and moderate hypoglycemia group [5.0 (4.0,5.5) mmol/Lvs.3.5 (3.0,3.9) mmol/L,3.3 (2.8,3.8) mmol/L] (P ＜ 0.05),but there was no significant difference in the onset of first hypoglycemia between groups (P ＞ 0.05).The aEEG score showed that there was significant difference in total score and sleep-wake cycle score between groups (P ＜ 0.05).The score of sleep-wake cycle in severe hypoglycemia group was significantly lower than that in moderate hypoglycemia group or in mild hypoglycemia group or in the control group (P ＜ 0.05),while there was no significant difference between moderate and mild hypoglycemia groups,and between moderate hypoglycemia and control group (P ＞ 0.05).Conclusion Severe hypoglycemia can lead to neonatal aEEG changes,mainly in the sleep-wake cycle changes.

[Event-related potential of working memory on emotional faces in children with autism spectrum disorder].

OBJECTIVE: To investigate the characteristics of working memory ability on emotional faces and related event-related potential (ERP) in children with autism spectrum disorder (ASD). METHODS: The Chinese Facial Affective Picture System was used as test material, and the event-related potential system was used to record the electroencephalographic data when 16 ASD children aged 6-12 years (ASD group) and 14 normal children matched for age (control group) were completing the facial emotion delayed match-to-sample task. The characteristics of P3b component were analyzed for both groups. RESULTS: Compared with the control group, the ASD group had a significantly longer reaction time (1 527 ms vs 1 060 ms; P<0.05) and a significantly lower accuracy rate (76% vs 88%; P<0.01) in the facial emotion delayed match-to-sample task. There was a difference in the amplitude of P3b component during the encoding stage between the two groups. In the ASD group, the P3b component on the left side electrode had a higher amplitude than that on the right side electrode (P<0.05), while the control group had no such characteristics. CONCLUSIONS: There is a difference in P3b component during the encoding stage between school-aged ASD children and normal children. In ASD children, working memory on emotional faces may depend more on the related neural pathway in the left hemisphere.

Event-related potential of working memory on emotional faces in children with autism spectrum disorder / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the characteristics of working memory ability on emotional faces and related event-related potential (ERP) in children with autism spectrum disorder (ASD).</p><p><b>METHODS</b>The Chinese Facial Affective Picture System was used as test material, and the event-related potential system was used to record the electroencephalographic data when 16 ASD children aged 6-12 years (ASD group) and 14 normal children matched for age (control group) were completing the facial emotion delayed match-to-sample task. The characteristics of P3b component were analyzed for both groups.</p><p><b>RESULTS</b>Compared with the control group, the ASD group had a significantly longer reaction time (1 527 ms vs 1 060 ms; P<0.05) and a significantly lower accuracy rate (76% vs 88%; P<0.01) in the facial emotion delayed match-to-sample task. There was a difference in the amplitude of P3b component during the encoding stage between the two groups. In the ASD group, the P3b component on the left side electrode had a higher amplitude than that on the right side electrode (P<0.05), while the control group had no such characteristics.</p><p><b>CONCLUSIONS</b>There is a difference in P3b component during the encoding stage between school-aged ASD children and normal children. In ASD children, working memory on emotional faces may depend more on the related neural pathway in the left hemisphere.</p>

Translocation of PKG1α acts on TRPV4-C1 heteromeric channels to inhibit endothelial Ca(2+) entry.

AIM: TRPV4-C1 heteromeric channels contribute to store-operated Ca(2+) entry in vascular endothelial cells. However, the negative regulation of these channels is not fully understood. This study was conducted to investigate the inhibitory effect of PKG1α on TRPV4-C1 heteromeric channels. METHODS: Immuno-fluorescence resonance energy transfer (FRET) was used to explore the spatial proximity of PKG1α and TRPC1. Phosphorylation of endogenous TRPC1 was tested by phosphorylation assay. [Ca(2+)]i transients and cation current in MAECs were assessed with Fura-2 fluorescence and whole-cell recording, respectively. In addition, rat mesenteric arteries segments were prepared, and vascular relaxation was examined with wire myography. RESULTS: In immuno-FRET experiments, after exposure of these cells to 8-Br-cGMP, more PKG1α was observed in the plasma membrane, and PKG1α and TRPC1 were observed to be in closer proximity. TAT-TRPC1(S172) and TAT-TRPC1(T313) peptide fragments, which contain the PKG targeted residues Ser172 and Thr313, respectively, were introduced into isolated endothelial cells to abrogate the translocation of PKG1α. Furthermore, a phosphorylation assay demonstrated that PKG directly phosphorylates TRPC1 at Ser172 and Thr313 in endothelial cells. In addition, PKG activator 8-Br-cGMP markedly reduced the magnitude of the 4αPDD-induced and 11,12-EET-induced [Ca(2+)]i transients, the cation current and vascular relaxation. CONCLUSION: This study uncovers a novel mechanism by which PKG negatively regulates endothelial heteromeric TRPV4-C1 channels through increasing the spatial proximity of TRPV4-C1 to PKG1α via translocation and through phosphorylating Ser172 and Thr313 of TRPC1.

Study on triterpenoic acids distribution in Ganoderma mushrooms by automatic multiple development high performance thin layer chromatographic fingerprint analysis.

Ganoderma--"Lingzhi" in Chinese--is one of the superior Chinese tonic materia medicas in China, Japan, and Korea. Two species, Ganoderma lucidum (Red Lingzhi) and G. sinense (Purple Lingzhi), have been included in the Chinese Pharmacopoeia since its 2000 Edition. However, some other species of Ganoderma are also available in the market. For example, there are five species divided by color called "Penta-colors Lingzhi" that have been advocated as being the most invigorating among the Lingzhi species; but there is no scientific evidence for such a claim. Morphological identification can serve as an effective practice for differentiating the various species, but the inherent quality has to be delineated by chemical analysis. Among the diverse constituents in Lingzhi, triterpenoids are commonly recognized as the major active ingredients. An automatic triple development HPTLC fingerprint analysis was carried out for detecting the distribution consistency of the triterpenoic acids in various Lingzhi samples. The chromatographic conditions were optimized as follows: stationary phase, precoated HPTLC silica gel 60 plate; mobile phase, toluene-ethyl acetate-methanol-formic acid (15 + 15 + 1 + 0.1); and triple-development using automatic multiple development equipment. The chromatograms showed good resolution, and the color images provided more specific HPTLC fingerprints than have been previously published. It was observed that the abundance of triterpenoic acids and consistent fingerprint pattern in Red Lingzhi (fruiting body of G. lucidum) outweighs the other species of Lingzhi.

Chemical pattern-aided classification to simplify the intricacy of morphological taxonomy of Epimedium species using chromatographic fingerprinting.

Epimedium herb (Yinyanghuo), one of the popular Chinese materia medica, is a multiple species colony of Epimedium genus belonging to Berberidaceae. There are five species of Epimedium that have been officially adopted in Chinese Pharmacopoeia under the same crude drug name 'Yinyanghuo' comprising Epimedium brevicornu, E. koreanum, E. sagittatum, E. pubescens, and E. wushanense. In addition, non-official species like E. acuminatum, E. miryanthum and E. leptorrhizum are also mix-used. Frequently, the morphological taxonomical identification is very difficult during on-site inspection for species authentication in the market. Researchers are often bewildered by the multiple species ambiguity when putting this crude drug in use. Referring to the bioactive constituents that are vital for therapeutic efficacy, the key to clarifying the multiple species confusion should rely on analysis of the bioactive composition. It is well known that medicinal Epimedium herbs contain special C-8 prenylated flavonol glycosides which contribute to various bioactivities and the major four, epimedin A (A), epimedin B (B), epimedin C (C) and icariin (I), are unanimously used as bioactive markers for quality control. In this study, HPLC-DAD fingerprinting was performed for investigating the molecular spectrum of various Epimedium species. It was found that the four major flavonoids constitute the middle part of the chromatographic profiles to form a specific region (named as 'ABCI fingerprint region') being dominant in the HPLC profiles of all medicinal Epimedium species, and the five official species express five different 'ABCI' patterns (different peak: peak ratios). Our study found that the convergent tendency of the 'ABCI region' among multiple species of Epimedium could facilitate differentiation of complex commercial samples based on similar bioactive composition should confer similar bioactivities. Merging the different species that possess the same 'ABCI region' pattern into the same group can create a simpler bioactive-fraction-aided classification array by clustering the commercial samples into three bioactive ingredients-based fingerprint patterns - 'E.b. pattern', 'E.k. pattern' and 'extensive E.w. pattern'. This approach offers the feasibility of characterizing and quality-controlling complex samples in the same genus designated under a single herbal drug entity on the premise of possessing the same bioactive ingredients pattern and supported by long-term traditional usage.

Study of the destructive effect to inherent quality of Angelicae dahuricae radix (Baizhi) by sulfur-fumigated process using chromatographic fingerprinting analysis.

The after-harvesting sun-dried process of Angelicae dahuricae radix (Chinese name: Baizhi) was previously the traditional treatment for commodity. Over recent decades the natural drying process for some fleshy roots or rhizomes of Chinese materia medica has been replaced by sulfur-fumigation for curtailing the drying duration and pest control. We used high performance liquid chromatography (HPLC) and high performance thin-layer chromatography (HPTLC) fingerprinting analysis to investigate the potential damaging effect of the sulfur-fumigating process. The experimental conditions were as follows. HPTLC analysis was carried out on pre-coated silica-gel 60 plate, twice development was performed with two solvent systems (mobile phase) A, chloroform-ethyl acetate (10:1) and B, hexane-chloroform-ether (4:1:2); the fluorescent images were observed under UV 365 nm. HPLC was preceeded on Zorbax SB-C(18) column; the linear gradient elution was conducted with mobile phase prepared from methanol-0.5% acetic acid; column temperature was at 25 degrees C; the detection wavelength was 250 nm. We found serious degradation of the majority of coumarins in sulfur-fumigated Baizhi. The destructive effect was manifested by the defaced chromatographic profile and verified by imitating the sulfur dioxide reaction with the constituents in Baizhi in the laboratory. It is suggested that sulfur-fumigation process is an unacceptable approach for processing herbal drugs.

[Study of HPLC-DAD fingerprint on complex traditional Chinese medicine proprietary preparation-Baoji pills].

OBJECTIVE: Based on 'Back-tracking' method, identification and quality evaluation of complex traditional Chinese medicine (TCM) preparation of Baoji pills (BJP) were carried out by HPLC fingerprint analysis. METHOD: HPLC-DAD fingerprint of BJP was conducted with Zorbax SB-C18 column and non-linear elution with the mobile phase consisted of acetonitrile-0.5% glacial acetic acid at column temperature 30 degrees C and detective wavelengths of 250 nm and 283 nm. From the established chromatographic pattern of BJP, track backward to the corresponding crude herbal drugs in the formula, attribution ofmost peaks in the BJP fingerprint can be disclosed. RESULT: The BJP HPLC fingerprint consisted of 44 peaks among which 35 peaks were assigned by parallel comparison with the fingerprint of the 10 corresponding crude drugs in the formula such as pueraria, pummelo peel, and magnolia bark, etc. and 22 peaks we reidentified by comparison with the chemical reference substances. CONCLUSION: The established HPLC fingerprint represents the whole character of BJP, which enhanced the specialty for control and assessment of the product quality. It exemplified much more effective for quality control than selecting any marker for qualitative or quantitative testing target. And the Back-tracking' experimental method extended the study mentality for complex formula TCM products chromatographic fingerprinting analysis.

Study of HPLC-DAD fingerprint on complex traditional Chinese medicine proprietary preparation-Baoji pills / 中国中药杂志

<p><b>OBJECTIVE</b>Based on 'Back-tracking' method, identification and quality evaluation of complex traditional Chinese medicine (TCM) preparation of Baoji pills (BJP) were carried out by HPLC fingerprint analysis.</p><p><b>METHOD</b>HPLC-DAD fingerprint of BJP was conducted with Zorbax SB-C18 column and non-linear elution with the mobile phase consisted of acetonitrile-0.5% glacial acetic acid at column temperature 30 degrees C and detective wavelengths of 250 nm and 283 nm. From the established chromatographic pattern of BJP, track backward to the corresponding crude herbal drugs in the formula, attribution ofmost peaks in the BJP fingerprint can be disclosed.</p><p><b>RESULT</b>The BJP HPLC fingerprint consisted of 44 peaks among which 35 peaks were assigned by parallel comparison with the fingerprint of the 10 corresponding crude drugs in the formula such as pueraria, pummelo peel, and magnolia bark, etc. and 22 peaks we reidentified by comparison with the chemical reference substances.</p><p><b>CONCLUSION</b>The established HPLC fingerprint represents the whole character of BJP, which enhanced the specialty for control and assessment of the product quality. It exemplified much more effective for quality control than selecting any marker for qualitative or quantitative testing target. And the Back-tracking' experimental method extended the study mentality for complex formula TCM products chromatographic fingerprinting analysis.</p>