ObjectiveTo evaluate the clinical efficacy and safety of acupuncture in treating severe Bell's palsy and to explore its potential mechanism by investigating the effect on serum levels of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF）. MethodsA randomized， subject-blinded， sham-acupuncture controlled trial was conducted. A total of 130 patients with severe Bell's palsy were randomly allocated into a treatment group or a control group at a 1∶1 ratio. Both groups received conventional western medicine. In addition， the treatment group received acupuncture， while the control group received sham acupuncture， with each session lasting 30 minutes. The treatment course lasted 8 weeks for both groups， followed by a follow-up assessment at week 12. The primary outcome was the proportion of patients achieving House-Brackmann （H-B） grade Ⅱ or lower at week 8. Secondary outcomes included Sunnybrook facial grading system scores at week 0， 4， 8， and 12， the time to satisfactory recovery（the time required to achieve H-B grade≤Ⅱ）， distribution of H-B grades and facial disability index （FDI） scores including the physical function subscale （FDIP） and social/well-being function subscale （FDIS） scores at week 0， 4， 8， and 12， and serum GDNF and NGF levels at week 0， 4， and 8. Adverse events and participants' self-assessments of treatment efficacy were also recorded. ResultsA total of 122 participants completed the study， including 62 in the treatment group and 60 in the control group. An intention-to-treat （ITT） analysis was performed， and missing data were handled using the last observation carried forward （LOCF） method. The proportion of patients achieving H-B grade ≤grade Ⅱ at week 8 was 78.5% （51/65） in the treatment group， significantly higher than 49.2% （32/65） in the control group （P＜0.05）. The Sunnybrook scores， FDIP and FDIS scores increased， while H-B grades decreased at week 4， 8， and 12 in both groups compared to week 0； moreover， improvements in all outcome measures were significantly greater in the treatment group than in the control group （P＜0.05）. The median time to satisfactory recovery was 6 weeks （95%CI： 5.697-6.303） in the treatment group， significantly shorter than 12 weeks （95%CI： 8.314-15.686） in the control group （P＜0.05）. Serum levels of GDNF and NGF were significantly higher in the treatment group at weeks 4 and 8 （P＜0.05）. No serious acupuncture-related adverse events occurred in either group. Adverse events were reported in 5 patients （7.69%） in the treatment group and 4 patients （6.15%） in the control group， with no statistically significant difference between groups （P＞0.05）. Patients' self-assessment of treatment efficacy after 8 weeks treatment was significantly better in the treatment group （P＜0.05）. ConclusionAcupuncture can effectively improve facial nerve function and shorten recovery time in patients with severe Bell's palsy， with a favorable safety profile. The therapeutic mechanism may be associated with the upregulation of serum GDNF and NGF levels.

To investigate the efficacy of hetrombopag for thrombocytopenia in patients with idiopathic multicentric Castleman disease(iMCD)-TAFRO(thrombocytopenia, anasarca, fever, reticulin fibrosis, renal dysfunction, and organomegaly)subtype.

Objective: To investigate the therapeutic effect of patchouli alcohol on mice with lung-heat syndrome based on the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-kappa B(NF-κB) signaling pathway. Methods: First, network pharmacology was used to predict the potential targets of patchouli alcohol in the treatment of lung-heat syndrome, and a "component-disease-key target" network was constructed for pathway analysis. Then, 40 BALB/c mice were assigned to the normal, lung-heat model, honeysuckle, and low-dose and high-dose patchouli alcohol groups. All groups, except the blank group, were intranasally infected with 50 μL (103 TCID50) of influenza virus solution. After two hours of infection, mice were treated once a day for seven consecutive days. The therapeutic mechanism of patchouli alcohol was explored by measuring pulmonary inflammatory factors, the PI3K/Akt/NF-κB pathway, hypothalamic fever markers (PGE2, cAMP, cGMP levels), rectal temperature, and tissue energy metabolism. Results: Network pharmacology identified 135 target genes related to patchouli alcohol and lung-heat syndrome, with the key targets being STAT3, H1F1A, and NF-κB1. In animal experiments, patchouli alcohol significantly alleviated influenza virus-induced lung inflammatory damage in mice with lung-heat syndrome, inhibited the expression of TNF-α and IL-6 in lung tissues(P<0.01), and suppressed the activation of the PI3K/Akt/NF-κB pathway. It also reduced hypothalamic levels of PGE2 and cAMP(P<0.01), suppressed the increase in rectal temperature, significantly decreased liver glycogen and pyruvate levels(P<0.01), and increased the activities of SDH, LDH, and Na+ -K+ -ATPase in the liver(P<0.01) Conclusion Patchouli alcohol improves the symptoms of lung-heat syndrome in mice by inhibiting the activation of the PI3K/Akt/NF-κB pathway, reducing proinflammatory cytokines and inflammatory damage, and regulating hypothalamic fever markers and energy metabolism.

Objective:To investigate the clinical manifestations and genetic characteristics of a Chinese Han family with Axenfeld-Rieger syndrome (ARS).Methods:A pedigree study was conducted.Three people from a Chinese Han family with ARS who visited Henan Eye Hospital in January 2024 were included, including 1 patient.Clinical data of the proband and her parents were collected.Comprehensive ophthalmic examination and general physical examination were performed on the proband and her parents.Peripheral blood samples were obtained from family members for DNA extraction.Whole exome sequencing was performed on the proband, and the copy number of the ZBED1P1, ENPEP, PITX2, and FAM241A genes in family members were validated using the real-time fluorescent quantitative PCR.Axenfeld-Rieger syndrome, Axenfeld-Rieger Syndrome, and PITX2 were used as keywords to search across databases such as OMIM, ClinVar, PubMed, CNKI, Wanfang, VIP, DECIPHER, and Google Scholar.The clinical manifestations and microdeletion types of different patients in ARS literature related to PITX2 microdeletions in China population were summarized, and the relationship between genotype and clinical phenotype was analyzed.The study followed the Declaration of Helsinki, and the study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEEC-2024[34]).All subjects understood the purpose of the study and voluntarily signed the informed consent form. Results:The proband was a 25-year-old female, exhibiting diminutive cornea in both eyes, polycoria, deformation and displacement of pupils, a flat mid-face, maxillary dysplasia, tooth loss, and a protruding umbilicus, among other symptoms.Parents of the proband were phenotypically normal.DNA sequencing identified a 1.06 MB microdeletion on chromosome 4q25 in the proband.Real-time quantitative PCR confirmed that this microdeletion encompassed the PITX2 and ENPEP genes, and it was absent in the proband's parents.The ClinGen CNV pathogenicity scoring indicated that the deletion involving the PITX2 gene represented a novel pathogenic copy number variation (CNV).Five studies related to 4q25 microdeletion in Chinese families with Axenfeld-Rieger syndrome was screened, including 13 patients.Clinical manefestations of the 13 patients included corneal disorders (accounting for 100%), umbilical hernia and dental anomalies (accounting for 92%), irregular intraocular pressure (accounting for 62%), iris atrophy (accounting for 46%), and posterior corneal embryotoxon (accounting for 31%). Conclusions:For this Chinese family diagnosed with ARS, a novel pathogenic 4q25 microdeletion variant encompassing the PITX2 gene was found in the proband, which is associated with characteristic phenotypes including microcornea, congenital iris dysplasia, polycoria, tooth loss, and a protruding umbilicus.

[Objective]To summarize the discussion on the bitter in Internal Classic of Huangdi and further explore and develop the theory of the bitter with later generations of medical practitioners,in order to bring in a new perspective for the further study.[Methods]Sorting out the viewpoints of Internal Classic of Huangdi and elaborating the original meaning of bitter and its derivatives by retrieving the database and combining it with the origin text of Internal Classic of Huangdi,it summarizes the cognition of bitter medicine,function,compatibility and contraindication of prescription and relavant contents of the bitter,making relative theory of bitter move clearly.[Results]Internal Classic of Huangdi deduces that bitter mainly encompasses the bitter of the five flavors,mouth bitterness in pathology,exertion bitterness and disease bitterness,and the bitter of the five flavors is the main content,which mainly includes the attributes of Yin and Yang of the bitterness,the efficacy,the contraindications of the combination of the bitter and the relationship with the five viscera.Bitter is considered as Yin,therefore,it can discharge,lower and be firm;the bitter taste is also produced by fire,so it can dry,warm and emit.The compatibility of bitter taste includes the methods of spicy opening bitter lowering,salty cold and sweet bitter,and bitter hot and sour bland.When combined with the other five flavors,it can benefit the heart,drain the lungs,firm the kidneys,help the liver and strengthen the spleen.[Conclusion]Internal Classic of Huangdi's bitter flavour laid the foundation for the theory of five flavors in later generations.Mastering the medicinal properties and effects of bitter flavour can lead to more appropriate and flexible clinical prescriptions.

Objective:To investigate the clinical manifestations and genetic characteristics of a Chinese Han family with Axenfeld-Rieger syndrome (ARS).Methods:A pedigree study was conducted.Three people from a Chinese Han family with ARS who visited Henan Eye Hospital in January 2024 were included, including 1 patient.Clinical data of the proband and her parents were collected.Comprehensive ophthalmic examination and general physical examination were performed on the proband and her parents.Peripheral blood samples were obtained from family members for DNA extraction.Whole exome sequencing was performed on the proband, and the copy number of the ZBED1P1, ENPEP, PITX2, and FAM241A genes in family members were validated using the real-time fluorescent quantitative PCR.Axenfeld-Rieger syndrome, Axenfeld-Rieger Syndrome, and PITX2 were used as keywords to search across databases such as OMIM, ClinVar, PubMed, CNKI, Wanfang, VIP, DECIPHER, and Google Scholar.The clinical manifestations and microdeletion types of different patients in ARS literature related to PITX2 microdeletions in China population were summarized, and the relationship between genotype and clinical phenotype was analyzed.The study followed the Declaration of Helsinki, and the study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEEC-2024[34]).All subjects understood the purpose of the study and voluntarily signed the informed consent form. Results:The proband was a 25-year-old female, exhibiting diminutive cornea in both eyes, polycoria, deformation and displacement of pupils, a flat mid-face, maxillary dysplasia, tooth loss, and a protruding umbilicus, among other symptoms.Parents of the proband were phenotypically normal.DNA sequencing identified a 1.06 MB microdeletion on chromosome 4q25 in the proband.Real-time quantitative PCR confirmed that this microdeletion encompassed the PITX2 and ENPEP genes, and it was absent in the proband's parents.The ClinGen CNV pathogenicity scoring indicated that the deletion involving the PITX2 gene represented a novel pathogenic copy number variation (CNV).Five studies related to 4q25 microdeletion in Chinese families with Axenfeld-Rieger syndrome was screened, including 13 patients.Clinical manefestations of the 13 patients included corneal disorders (accounting for 100%), umbilical hernia and dental anomalies (accounting for 92%), irregular intraocular pressure (accounting for 62%), iris atrophy (accounting for 46%), and posterior corneal embryotoxon (accounting for 31%). Conclusions:For this Chinese family diagnosed with ARS, a novel pathogenic 4q25 microdeletion variant encompassing the PITX2 gene was found in the proband, which is associated with characteristic phenotypes including microcornea, congenital iris dysplasia, polycoria, tooth loss, and a protruding umbilicus.

Objective:To analyze the effect of nucleotide sequence variants in the 5′ untranslated region (5′UTR) of Helicobacter pylori (Hp) cagA on mRNA secondary structure, as well as its regulatory role in cytotoxin-associated gene A (CagA) protein expression and bacterial virulence. Methods:The upstream nucleotide sequence of cagA was amplified by polymerase chain reaction (PCR) from 37 Hp strains, and the PCR products were sequenced. MEGA 5.0 software and RNAfold prediction software were used to analyze the nucleotide sequence variants of cagA 5′UTR and the changes in mRNA secondary structure of this region, respectively. Western blot was used to detect the expression level of CagA protein in Hp strains, and the regulatory effect of cagA 5′UTR variants on the difference in CagA protein expression was analyzed. An Hp-infected AGS cell model was established to evaluate bacterial adhesion rate; quantitative PCR (qPCR) was used to analyze the mRNA transcription levels of interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α); enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion levels of IL-8 and TNF-α proteins. Results:Nucleotide sequence alignment of cagA 5′UTR from 37 Hp strains showed that sequence differences were mainly concentrated in the -53motif, -10motif, + 34motif, and + 86motif regions. mRNA secondary structure prediction analysis revealed three types based on the StemB stem-loop structure: type Ⅰ (no StemB stem-loop), type ⅡA (StemB stem-loop with 3-4 base partial pairing), and type ⅡB (StemB stem-loop with 5 base full pairing). Western blot analysis showed that the CagA protein expression level was the highest in type Ⅰ Hp strains (1.72±0.29) and the lowest in type ⅡB strains (0.81±0.26), with a statistically significant difference between the two types ( P=0.030). The adhesion rate of type Ⅰ Hp strains to AGS cells was (52.90±11.17)%, which was higher than that of type Ⅱ strains [(21.27±6.16)%]. qPCR results showed that the mRNA transcription levels of IL-8 and TNF-α in AGS cells induced by type Ⅰ Hp strains were higher than those induced by type Ⅱ strains (140.23±24.47 vs 76.16±8.76, P=0.069; 55.20±9.04 vs 21.26±6.16, P=0.036). ELISA analysis further indicated that the secretion levels of IL-8 and TNF-α proteins in AGS cells induced by type Ⅰ Hp strains were also higher than those induced by type Ⅱ strains [(344.66±62.62)pg/ml vs (302.13±66.27)pg/ml, P=0.665; (131.04±4.94)pg/ml vs (79.17±11.32)pg/ml, P=0.014]. Conclusions:The cagA 5′UTR region of Hp strains exhibits significant nucleotide sequence variants. Hp strains with no StemB stem-loop (type Ⅰ) in the mRNA secondary structure show significantly increased CagA protein expression and higher bacterial pathogenic potential.

Objective:To analyze the effect of nucleotide sequence variants in the 5′ untranslated region (5′UTR) of Helicobacter pylori (Hp) cagA on mRNA secondary structure, as well as its regulatory role in cytotoxin-associated gene A (CagA) protein expression and bacterial virulence. Methods:The upstream nucleotide sequence of cagA was amplified by polymerase chain reaction (PCR) from 37 Hp strains, and the PCR products were sequenced. MEGA 5.0 software and RNAfold prediction software were used to analyze the nucleotide sequence variants of cagA 5′UTR and the changes in mRNA secondary structure of this region, respectively. Western blot was used to detect the expression level of CagA protein in Hp strains, and the regulatory effect of cagA 5′UTR variants on the difference in CagA protein expression was analyzed. An Hp-infected AGS cell model was established to evaluate bacterial adhesion rate; quantitative PCR (qPCR) was used to analyze the mRNA transcription levels of interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α); enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion levels of IL-8 and TNF-α proteins. Results:Nucleotide sequence alignment of cagA 5′UTR from 37 Hp strains showed that sequence differences were mainly concentrated in the -53motif, -10motif, + 34motif, and + 86motif regions. mRNA secondary structure prediction analysis revealed three types based on the StemB stem-loop structure: type Ⅰ (no StemB stem-loop), type ⅡA (StemB stem-loop with 3-4 base partial pairing), and type ⅡB (StemB stem-loop with 5 base full pairing). Western blot analysis showed that the CagA protein expression level was the highest in type Ⅰ Hp strains (1.72±0.29) and the lowest in type ⅡB strains (0.81±0.26), with a statistically significant difference between the two types ( P=0.030). The adhesion rate of type Ⅰ Hp strains to AGS cells was (52.90±11.17)%, which was higher than that of type Ⅱ strains [(21.27±6.16)%]. qPCR results showed that the mRNA transcription levels of IL-8 and TNF-α in AGS cells induced by type Ⅰ Hp strains were higher than those induced by type Ⅱ strains (140.23±24.47 vs 76.16±8.76, P=0.069; 55.20±9.04 vs 21.26±6.16, P=0.036). ELISA analysis further indicated that the secretion levels of IL-8 and TNF-α proteins in AGS cells induced by type Ⅰ Hp strains were also higher than those induced by type Ⅱ strains [(344.66±62.62)pg/ml vs (302.13±66.27)pg/ml, P=0.665; (131.04±4.94)pg/ml vs (79.17±11.32)pg/ml, P=0.014]. Conclusions:The cagA 5′UTR region of Hp strains exhibits significant nucleotide sequence variants. Hp strains with no StemB stem-loop (type Ⅰ) in the mRNA secondary structure show significantly increased CagA protein expression and higher bacterial pathogenic potential.

[Objective]To summarize the discussion on the bitter in Internal Classic of Huangdi and further explore and develop the theory of the bitter with later generations of medical practitioners,in order to bring in a new perspective for the further study.[Methods]Sorting out the viewpoints of Internal Classic of Huangdi and elaborating the original meaning of bitter and its derivatives by retrieving the database and combining it with the origin text of Internal Classic of Huangdi,it summarizes the cognition of bitter medicine,function,compatibility and contraindication of prescription and relavant contents of the bitter,making relative theory of bitter move clearly.[Results]Internal Classic of Huangdi deduces that bitter mainly encompasses the bitter of the five flavors,mouth bitterness in pathology,exertion bitterness and disease bitterness,and the bitter of the five flavors is the main content,which mainly includes the attributes of Yin and Yang of the bitterness,the efficacy,the contraindications of the combination of the bitter and the relationship with the five viscera.Bitter is considered as Yin,therefore,it can discharge,lower and be firm;the bitter taste is also produced by fire,so it can dry,warm and emit.The compatibility of bitter taste includes the methods of spicy opening bitter lowering,salty cold and sweet bitter,and bitter hot and sour bland.When combined with the other five flavors,it can benefit the heart,drain the lungs,firm the kidneys,help the liver and strengthen the spleen.[Conclusion]Internal Classic of Huangdi's bitter flavour laid the foundation for the theory of five flavors in later generations.Mastering the medicinal properties and effects of bitter flavour can lead to more appropriate and flexible clinical prescriptions.

Objective:To analyze the clinical phenotypes and pathogenic gene of a Han Chinese family with enhanced S-cone syndrome (ESCS).Methods:The method of pedigree investigation was adopted.A suspected ESCS Han Chinese family including 8 members of 3 generations was recruited in Henan Eye Hospital from June to September 2021.There was one patient in the family.A thorough ophthalmic examination of the proband was carried out to evaluate the phenotypes, including visual acuity, degree of strabismus, anterior segment and fundus, autofluorescence imaging, fluorescein fundus angiography, full-field electroretinogram (ERG), multifocal ERG, optical coherence tomography.DNA was extracted from peripheral blood samples from the proband and family members.The pathogenic gene and variation were screened by whole exome sequencing (WES).The variation and co-segregation were verified by Sanger sequencing.The deleteriousness of the variation was analyzed by SIFT, Polyphen2 and MutationTaster.The pathogenicity of the variation was evaluated in accordance with the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines.The analysis of amino acid sequence conservation was performed by SIFT.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2017[6]).Written informed consent was obtained from each subject.Results:This pedigree was consistent with autosomal recessive inheritance.The proband had clinical features such as night blindness, hyperopia, accommodative esotropia, peripheral retinal pigmentation, retinoschisis, and photopic ERG responses dominated by large-amplitude waves.Variations including a compound heterozygous variation, c.671C>T: p.S224L on exon 5 and c. 955G>A: p.E319K on exon 6 of NR2E3 were identified by WES.The variations were confirmed to be consistent with co-segregation.The both loci were missense variations, the variation frequency of which was 0 in the East Asian population via the gnomAD database.The variations were predicted to be deleterious by SIFT, Polyphen2 and MutationTaster.The c.671C>T variation was recorded with unknown significance in ClinVar database, and the c.955G>A variation was an unreported new locus.According to the ACMG Standards and Guidelines, the both variations were labeled as with uncertain clinical significance, and the corresponding amino acid sequences were highly conservative across multiple species. Conclusions:This family has the clinical characteristics of ESCS and meets the genetic diagnosis criteria.Two novel variations in NR2E3 gene, c.671C>T: p.S224L and 955G>A: p.E319K, are found.