In vitro anti-Helicobacter pylori activity and the underlining mechanism of an empirical herbal formula - Hezi Qingyou.

Background: Helicobacter pylori (H. pylori) is thought to primarily colonize the human stomach and lead to various gastrointestinal disorders, such as gastritis and gastric cancer. Currently, main eradication treatment is triple or quadruple therapy centered on antibiotics. Due to antibiotic resistance, the eradication rate of H. pylori is decreasing gradually. Therefore, searching for anti-H. pylori drugs from herbal sources has become a strategy for the treatment. Our team proposed a Hezi Qingyou Formula (HZQYF), composed of Chebulae Fructus, Ficus hirta Vahl and Cloves, and studied its anti-H. pylori activity and mechanism. Methods: Chemical components of HZQYF were studied using UHPLC-MS/MS and HPLC. Broth microdilution method and agar dilution method were used to evaluate HZQYF's antibacterial activity. The effects of HZQYF on expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF), and flagellar genes (flaA, flaB) were explored using Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) technology. Effects on morphology and permeability of the extracellular membrane were studied using scanning electron microscopy (SEM) and N-phenylnaphthalen-1-amine (NPN) uptake. Effect on urease activity was studied using a urease kinetics analysis in vitro. Immunofluorescence staining method was used to examine the effect on adhesion. Western blot was used to examine the effect on cagA protein. Results: Minimum inhibitory concentration (MIC) values of the formula against H. pylori clinical strains and standard strains were 80-160 µg/mL, and minimum bactericidal concentration (MBC) values were 160-320 µg/mL. The formula could down-regulate the expression of adhesion genes (alpA, alpB, babA), urease genes (ureE, ureF) and flagellar genes (flaA, flaB), change the morphology of H. pylori, increase its extracellular membrane permeability, and decrease its urease activity. Conclusion: Present studies confirmed that HZQYF had promising in vitro anti-H. pylori activities and demonstrated its possible mechanism of action by down-regulating the bacterial adhesion, urease, and flagellar gene expression, which provided scientific bases for further clinical investigations.

1,3,6-Trigalloylglucose: A Novel Potent Anti-Helicobacter pylori Adhesion Agent Derived from Aqueous Extracts of Terminalia chebula Retz.

1,3,6-Trigalloylglucose is a natural compound that can be extracted from the aqueous extracts of ripe fruit of Terminalia chebula Retz, commonly known as "Haritaki". The potential anti-Helicobacter pylori (HP) activity of this compound has not been extensively studied or confirmed in scientific research. This compound was isolated using a semi-preparative liquid chromatography (LC) system and identified through Ultra-high-performance liquid chromatography-MS/MS (UPLC-MS/MS) and Nuclear Magnetic Resonance (NMR). Its role was evaluated using Minimum inhibitory concentration (MIC) assay and minimum bactericidal concentration (MBC) assay, scanning electron microscope (SEM), inhibiting kinetics curves, urea fast test, Cell Counting Kit-8 (CCK-8) assay, Western blot, and Griess Reagent System. Results showed that this compound effectively inhibits the growth of HP strain ATCC 700392, damages the HP structure, and suppresses the Cytotoxin-associated gene A (Cag A) protein, a crucial factor in HP infection. Importantly, it exhibits selective antimicrobial activity without impacting normal epithelial cells GES-1. In vitro studies have revealed that 1,3,6-Trigalloylglucose acts as an anti-adhesive agent, disrupting the adhesion of HP to host cells, a critical step in HP infection. These findings underscore the potential of 1,3,6-Trigalloylglucose as a targeted therapeutic agent against HP infections.

Effects of combined acupuncture and medication on hyperarousal state and serum copeptin in patients with chronic insomnia / 针灸推拿医学（英文版）

Objective:To observe the effect of combined acupuncture and medication on hyperarousal state and serum copeptin(CPT)in patients with chronic insomnia(CI),and to explore its possible mechanism of action.Methods:A total of 70 CI patients meeting the inclusion criteria were divided into an observation group and a control group by the random number table method,with 35 cases in each group.The control group was given estazolam tablets before bedtime,1 mg/time,once a day.The observation group was treated with additional Yi Nao An Shen acupuncture therapy(acupuncture for benefiting the brain and tranquillization)on the basis of the medication treatment,4 times a week.After 4 weeks of treatment,the Pittsburgh sleep quality index(PSQI)score,insomnia severity index(ISI)score,pre-sleep arousal scale(PSAS)score,hyperarousal scale(HAS)score,and the change in serum CPT level were compared between the two groups.Results:During the study,there were 2 dropout cases in the observation group and 1 dropout case in the control group.After treatment,the PSQI,ISI,PSAS,and HAS scores and the serum CPT level in both groups decreased compared with the same group before treatment,and the intra-group differences were statistically significant(P<0.05).After treatment,changes in each above scale score and the serum CPT level in the observation group were much more significant and were statistically different from those in the control group(P<0.05).Conclusion:Acupuncture plus medication can improve sleep quality,reduce the degree of insomnia,and regulate hyperarousal state in patients with CI,and its mechanism of action may be related to the down-regulation of serum CPT level.

Analysis on acupoint selection rules for impotence treated with acupuncture and moxibustion in ancient based on data mining technology / 中国针灸

Based on the data mining technology, the rules of acupoint selection and prescription were analyzed for impotence treated with acupuncture and moxibustion in ancient recorded in

Clinical observation on filiform fire-needling plus continuous passive motion therapy for frozen shoulder / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of filiform fire-needling plus continuous passive motion (CPM) therapy for frozen shoulder. Methods: A total of 72 patients were randomized into an observation group and a control group by the random number table method, with 36 cases in each group. Patients in the control group received celecoxib capsule plus CPM, while those in the observation group received filiform fire-needling plus CPM. The whole course of treatment lasted for 2 weeks in both groups. The visual analog scale (VAS), Constant-Murley score (CMS) and range of motion of shoulder joint were measured for therapeutic efficacy evaluation. Results: The total effective rate was 91.7％ in the observation group, higher than 72.2％ in the control group, and the between-group comparison showed statistical significance (P<0.05). After treatment, the VAS scores in the two groups dropped significantly, the CMS as well as the range of motion including abduction, forward flexion and extension were all increased significantly, and the intra-group comparisons showed statistical significance (all P<0.05). The betweengroup comparisons showed the improvements in these items in the observation group were more significant than those in the control group (all P<0.05). Conclusion: Filiform fire-needling plus CPM can produce more significant efficacy than celecoxib capsule plus CPM for frozen shoulder; it can alleviate pain, improve shoulder function and restore joint range of motion in such patients.

acupuncture and massage academic school with "the combination of five meridians" for post-stroke urinary incontinence / 中国针灸

The theory of "combination of five meridians" focuses on meridians and acupoints as well as compatibility and interrelationship of generation and restriction of five elements. By regulating corresponding meridians to diseased, the relationship of unbalancedis balanced, which can regulate body function and effectively treat diseases. Under the guide of theory "the combination of five meridians", according to etiology and pathogenesis of post-stroke urinary incontinence, wind, fire and phlegm, from three aspects of deficiency, excess and deficient root and excessive surface, under the treatment principle of "nourishing the mother if it is deficient, reducing the son if it is excessive", the original meridian, son meridian, mother meridian, conquering meridian and conquered meridian are selected while the methods of nourishing the mother, reducing the son, reinforcing the suppression and developing the weak are applied, so as to discuss the acupuncture treatment for post-stroke urinary incontinence, which can provide new clinical thoughts of acupuncture treatment for post-stroke urinary incontinence.

Prevalence of neutralizing antibodies against liver-tropic adeno-associated virus serotype vectors in 100 healthy Chinese and its potential relation to body constitutions / 中西医结合学报

Recombinant adeno-associated virus (rAAV) serotype 2, 3 and 8 vectors are the most promising liver-tropic AAV serotype vectors. Liver diseases are significant problems in China. However, to date, few studies on AAV neutralizing antibodies (Nabs) were working with the Chinese population or with the rAAV3 vectors. The present study aimed to determine the prevalence of Nabs in Chinese population against wild-type AAV2, AAV3 and AAV8 capsids as well as additional two AAV3 variants. In addition, we performed a preliminary analysis to investigate the potential influence of traditional Chinese medicine body constitutions on AAV Nabs. Our work demonstrated that the majority of healthy Chinese subjects were positive for AAV Nabs, with the order of AAV2>AAV3=AAVLK03>AAV8. There was no difference between: 1) AAV3 and its variants; 2) male and female subjects; and 3) different age cohorts (≤35, 36-50, and ≥51 years old). People in the Qi-deficiency constitution had significantly increased AAV8 Nabs than people in the Gentleness constitution. Our studies may have impact on the future clinical design of AAV-based gene therapy in the Chinese population.

Acupoints selection rules analysis of ancient acupuncture for urinary incontinence based on data mining technology / 中国针灸

Based on ancient literature of acupuncture in Canon of Chinese Medicine (4th edition), the articles regarding acupuncture for urinary incontinence were retrieved and collected to establish a database. By Weka data mining software, the multi-level association rules analysis method was applied to analyze the acupoints selection characteristics and rules of ancient acupuncture for treatment of urinary incontinence. Totally 356 articles of acupuncture for urinary incontinence were collected, involving 41 acupoints with a total frequency of 364. As a result, (1) the acupoints in the yin-meridian of hand and foot were highly valued, as the frequency of acupoints in yin-meridians was 2.6 times than that in yang-meridians, and the frequency of acupoints selected was the most in the liver meridian of foot-jueyin; (2) the acupoints in bladder meridian of foot-taiyang were also highly valued, and among three yang-meridians of foot, the frequency of acupoints in the bladder meridian of foot-taiyang was 54, accounting for 65.85% (54/82); (3) more acupoints selected were located in the lower limbs and abdomen; (4) specific acupoints in above meridians were mostly selected, presenting 73.2% (30/41) to the ratio of number and 79.4% (289/364) to the frequency, respectively; (5) Zhongji (CV 3), the front-mu point of bladder meridian, was seldom selected in the ancient acupuncture literature, which was different from modern literature reports. The results show that urinary incontinence belongs to external genitalia diseases, which should be treated from yin, indicating more yin-meridians be used and special acupoints be focused on. It is essential to focus inheritance and innovation in TCM clinical treatment, and applying data mining technology to ancient literature of acupuncture could provide classic theory basis for TCM clinical treatment.

Present study on antagonistic effect of acupoints compatibility / 中国针灸

Compatibility of acupoints is an important component of acupuncture prescription, and it is the key point to achieve the therapeutic effect of acupuncture. Antagonism and synergism are held as two possible results of acupoints compatibility. Recent literatures are summarized and analyzed in this article. And clinical trials and laboratorial studies indicate that antagonism is a result of acupoints compatibility. However, since the relative studies are not enough, extensive attentions are not aroused among researchers. Therefore, in the future studies, based on specificity of acupoints, interdisciplinary research methods should be adopted to explore the effect of antagonism of acupoint compatibility on enhancing the clinical therapeutic effect and expound the mechanism of acupuncture and moxibustion. It is considered to be a significant topic which is worth to be deeply studied.

Amplification of human IgG Fc gene and its secretory expression in eukaryotic cells / 中国免疫学杂志

Objective:The immunogenicity of DNA vaccine immunogenicity can be improved by fusing antigenic genes to IgG Fc fragment.It is critical for enhancing immunogenicity of DNA vaccine to construct the secreting eukaryotic expressing vector.The purpose of this study is to construct a secreting eukaryotic expressing vector ligated with IgG Fc gene by amplifying human IgG Fc fragment and fusing the fragment with human CD5 signal peptide sequence for highly efficient expression.Methods:Human lymphocytes were isolated from the tonsil obtained by removal surgery.The total RNA of lymphocytes was extracted using Trizol reagent.Human IgG Fc cDNA was amplified by RT-PCR from lymphocytes and then inserted into pMD-18T vector.The Fc encoding sequence fused with CD5 signal peptide(sp) sequence was constructed by cross PCR using Fc fragment and CD5sp sequence and cloned in pcDNA-CD5 plasmid as the template.CD5sp-Fc fragment was inserted into pcDNA3.1 expressing vector to construct the pcDNA-CD5sp-Fc plasmid.The plasmids were transfected into HEK293T cells mediated by calcium phosphate.Western blot was used to detect expressed Fc protein in cultural supermatant of the cells.Results:The amplified sequence of human IgG Fc was consistent with that previously published.The secretory expression of Fc in HEK293T cells was achieved and the expressing level reached 50 ?g/106 cells at 48 h culture after transfection.Conclusion:The human IgG Fc gene is amplified by RT-PCR methods and the secretory expression of Fc gene mediated by CD5 signal peptide in 293T cells is achieved.The results provide a experimental basis for further construction of antigen genes fused to IgG Fc fragment and investigation on DNA vaccines and Fc as a biological adjuvant.