Hydroxyectoine, a vital compatible solute, is widely utilized in cosmetics, food, pharmaceutical industries, and biologics. However, the current microbial fermentation methods for hydroxyectoine production face challenges including insufficient precursor supply and low yields. Therefore, developing engineering microbial strains capable of efficiently synthesizing hydroxyectoine is of great significance. In this study, we first constructed a high-yield ectoine-producing strain ECT04 by multi-copy integration of the ectoine synthesis genes ectABC into the pseudogene loci of Escherichia coli MG1655(DE3), achieving an ectoine titer of 6.03 g/L. Subsequently, we employed plasmids with varying copy numbers to express ectD from Chromohalobacter salexigens to enable the conversion for hydroxyectoine production. We further investigated the effects of promoter, co-substrate ɑ-ketoglutarate, Fe²⁺ concentration, and dissolved oxygen on hydroxyectoine synthesis. Through fed-batch fermentation in a 7-L bioreactor, we significantly enhanced the hydroxyectoine production efficiency, attaining a final titer of 8.58 g/L and a productivity of 0.24 g/(L·h). This work successfully achieved the de novo synthesis of hydroxyectoine in E. coli, laying a foundation for the efficient bioproduction of this compound.
Escherichia coli/genetics* ; Fermentation ; Amino Acids, Diamino/biosynthesis* ; Bioreactors/microbiology* ; Metabolic Engineering/methods* ; Chromohalobacter/genetics* ; Plasmids/genetics*

Objective:To investigate the histopathological subtype and clinical characteristics of vascular malformations in the head and neck.Methods:All patients with head and neck vascular malformations referred to Henan Provincial People′s Hospital between January 2021 and December 2023 were collected, and the clinicopathological features were analyzed by SPSS 26.0 statistical software.Results:A total of 675 patients were included, there were 293 male and 382 female patients with a mean onset age of (26.2±21.8）years and a mean diagnosis age of (31.8±20.6）years. Most vascular malformations were located in oral and maxillofacial region (52.1%, 352/675), with an average diameter of (28.0±19.8）mm. Venous malformations were the most common (71.4%, 482/675). There were 230 patients presented with simultaneous phenomenon, and of them the thrombus was most commonly noted (66.5%, 153/230). Venous malformations more frequently occurred in patients who were older than 20 years, extracranial in site, and mostly involved female patients with complications. Combined vascular malformation more frequently occurred in patients who were under 20 years old, extracranial in site, and mostly involved female patients. Capillary malformations more frequently occurred in patients under 20 years old, extracranial in site, and no complications occurred. Lymphatic vessel malformation more frequently occurred in patients who were under 20 years old, extracranial in site, female and with no complications. Arteriovenous malformations were more common in the intracranial and most of them were without any complications. Two cases (2/21) were associated with PIK3CA gene mutation. There were 109 patients with recurrence after initial treatment.Conclusions:Vascular malformations in the head and neck are more commonly found in the extracranial and maxillofacial regions, and venous malformations are the most common type; while arteriovenous malformations are more common in the intracranial site. Appropriate diagnosis is the premise of appropriate treatment.

Objective To investigate the effects of Huayu Xiaopi Decoction on the proliferation and migration of AGS cells;To explore its mechanism in treating precancerous lesions of gastric cancer.Methods AGS cells were divided into blank group,inhibitor group and Huayu Xiaopi Decoction high-,medium-and low-dosage groups.The blank group was cultured with 15%control serum,the inhibitor group was cultured with 10 μmol/L HIF-1α inhibitor,and the Huayu Xiaopi Decoction high-,medium-and low-dosage groups were cultured with 12%,6%and 3%drug containing serum,respectively.CCK-8 method was used to detect cell survival rate,cell migration ability was detected by scratch test,immunofluorescence was used to detect the expressions of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP)-2 and MMP-9 in AGS cells,the expressions of HIF-1α,COX-2,VEGF,VEGFR2,MMP-2,MMP-9 mRNA and HIF-1α,COX-2,VEGF,VEGFR2 proteins in AGS cells were detected by RT-PCR and Western blot.Results Compared with the blank group,the cell survival rate and migration rate were significantly decreased in the inhibitor group and each dosage of Huayu Xiaopi Decoction groups(P<0.05),the expressions of PCNA,MMP-2 and MMP-9 significantly decreased in the inhibitor group and Huayu Xiaopi Decoction high-and medium-dosage groups(P<0.05),the mRNA expressions of HIF-1α,COX-2,VEGF,VEGFR2,MMP-2,MMP-9 and the protein expression of HIF-1α,COX-2,VEGF,VEGFR2 were significantly decreased(P<0.05).Conclusion Huayu Xiaopi Decoction can inhibit the proliferation and migration of AGS cells,and its mechanism is related to the regulation of the expression of key molecules in HIF-1α/VEGF signaling pathway.

Objective:To investigate the histopathological subtype and clinical characteristics of vascular malformations in the head and neck.Methods:All patients with head and neck vascular malformations referred to Henan Provincial People′s Hospital between January 2021 and December 2023 were collected, and the clinicopathological features were analyzed by SPSS 26.0 statistical software.Results:A total of 675 patients were included, there were 293 male and 382 female patients with a mean onset age of (26.2±21.8）years and a mean diagnosis age of (31.8±20.6）years. Most vascular malformations were located in oral and maxillofacial region (52.1%, 352/675), with an average diameter of (28.0±19.8）mm. Venous malformations were the most common (71.4%, 482/675). There were 230 patients presented with simultaneous phenomenon, and of them the thrombus was most commonly noted (66.5%, 153/230). Venous malformations more frequently occurred in patients who were older than 20 years, extracranial in site, and mostly involved female patients with complications. Combined vascular malformation more frequently occurred in patients who were under 20 years old, extracranial in site, and mostly involved female patients. Capillary malformations more frequently occurred in patients under 20 years old, extracranial in site, and no complications occurred. Lymphatic vessel malformation more frequently occurred in patients who were under 20 years old, extracranial in site, female and with no complications. Arteriovenous malformations were more common in the intracranial and most of them were without any complications. Two cases (2/21) were associated with PIK3CA gene mutation. There were 109 patients with recurrence after initial treatment.Conclusions:Vascular malformations in the head and neck are more commonly found in the extracranial and maxillofacial regions, and venous malformations are the most common type; while arteriovenous malformations are more common in the intracranial site. Appropriate diagnosis is the premise of appropriate treatment.

Objective To investigate the effects of Huayu Xiaopi Decoction on the proliferation and migration of AGS cells;To explore its mechanism in treating precancerous lesions of gastric cancer.Methods AGS cells were divided into blank group,inhibitor group and Huayu Xiaopi Decoction high-,medium-and low-dosage groups.The blank group was cultured with 15%control serum,the inhibitor group was cultured with 10 μmol/L HIF-1α inhibitor,and the Huayu Xiaopi Decoction high-,medium-and low-dosage groups were cultured with 12%,6%and 3%drug containing serum,respectively.CCK-8 method was used to detect cell survival rate,cell migration ability was detected by scratch test,immunofluorescence was used to detect the expressions of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP)-2 and MMP-9 in AGS cells,the expressions of HIF-1α,COX-2,VEGF,VEGFR2,MMP-2,MMP-9 mRNA and HIF-1α,COX-2,VEGF,VEGFR2 proteins in AGS cells were detected by RT-PCR and Western blot.Results Compared with the blank group,the cell survival rate and migration rate were significantly decreased in the inhibitor group and each dosage of Huayu Xiaopi Decoction groups(P<0.05),the expressions of PCNA,MMP-2 and MMP-9 significantly decreased in the inhibitor group and Huayu Xiaopi Decoction high-and medium-dosage groups(P<0.05),the mRNA expressions of HIF-1α,COX-2,VEGF,VEGFR2,MMP-2,MMP-9 and the protein expression of HIF-1α,COX-2,VEGF,VEGFR2 were significantly decreased(P<0.05).Conclusion Huayu Xiaopi Decoction can inhibit the proliferation and migration of AGS cells,and its mechanism is related to the regulation of the expression of key molecules in HIF-1α/VEGF signaling pathway.

Based on the"You Gu Wu Yun"theory in traditional Chinese medicine(TCM),this paper believes that"Gu"in"You Gu Wu Yun"is extended to"state"from the perspective of"TCM state".In order to avoid the adverse reactions of TCM,the macro,meso,and micro three views should be used together,and macro,meso,and micro parameters should be integrated.We should also carefully identify the physiological characteristics,pathological characteristics,constitution,syndrome,and disease of human body by combining qualitative and quantitative method,highlighting the relationship between the prescription and the"state".The correspondence between prescription and the"state"will reduce the risk of adverse reactions of TCM.In this paper,we hope to focus on the guiding role of the"You Gu Wu Yun"theory in TCM research,to give full play to the characteristics and advantages of TCM,and to dialectically treat the role of TCM.

In the context of the complex and ever-changing spectrum of diseases, the traditional Chinese medicine compatibility of prescription and drugs is no longer able to fully meet the needs of clinical diagnosis and treatment. Therefore, this article is based on the diagnosis and treatment model of the combination of disease, syndrome, and symptom, combined with the development achievements of Western medicine, and explores the principles of formulating traditional prescriptions based on the combination of chief, deputy, assistant, and envoy. This article proposes a formulation principle of composing prescriptions with the diagnosis of syndrome as the chief, the diagnosis of disease as the deputy, the treatment of symptoms as the assistant, and the harmonization of medicine as the envoy. This forms a treatment plan with the core link of syndrome differentiation and treatment, disease differentiation and treatment, symptomatic treatment, detoxification, and efficacy enhancement. The purpose of this article is to address the current clinical challenges such as an increasing disease spectrum and the complexity of syndrome patterns and symptom clusters. It aims to provide new insights into traditional Chinese medicine clinical treatment plans and herbal formulation strategies, with the ultimate goal of improving the clinical effectiveness of traditional Chinese medicine.

The correspondence between prescription and syndrome is the advantage and characteristic of traditional Chinese medicine(TCM)treatment.However,the pathogenesis of clinical diseases is complex and the condition is changeable,and the clinical application is difficult to achieve the maximum effect under the existing cognition of the correspondence between prescription and syndrome.In this paper,the five categories of physiological characteristics,pathological characteristics,constitution,syndrome,and disease of the longitudinal classification of"TCM state"are introduced into the correspondence of prescription and syndrome.Under the vertical perspective of"TCM state",the theoretical connotation of the correspondence between prescription and syndrome is interpreted as"correspondence between prescription and state",namely correspondence of"prescription-physiological characteristics",correspondence of"prescription-pathological characteristics",correspondence of"prescription-constitution",correspondence of"prescription-syndrome",and correspondence of"prescription-disease".It is hoped to accurately grasp the corresponding connotation of the correspondence between prescription and syndrome,in order to deepen the clinical thinking mode of TCM.

Objective:To investigate the endoscopic combined serological diagnosis strategy for G1 and G2 gastric neuroendocrine neoplasms (G-NENs), and to evaluate the safety, short-term, and long-term efficacy of two endoscopic treatment procedures: endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD).Methods:This study retrospectively analyzed the clinical data of 100 consecutive patients with G-NENs who were hospitalized at the Cancer Hospital of the Chinese Academy of Medical Sciences from January 2011 to October 2023. These patients underwent endoscopic treatment, and propensity score matching (PSM) was used to compare clinicopathological characteristics, as well as short-term and long-term efficacy of lesions in the EMR group and ESD group before and after treatment.Results:Among the 100 patients with G-NENs, the median age was 54 years old. Before surgery, 29 cases underwent endoscopic combined serological examination, and 24 of them (82.2%) had abnormally elevated plasma chromogranin A. The combined diagnostic strategy for autoimmune atrophic gastritis (AIG) achieved a diagnostic accuracy of 100%(22/22). A total of 235 G-NEN lesions were included, with 84 in the ESD group and 151 in the EMR group. The median size of the lesions in the ESD group (5.0 mm) was significantly larger than that in the EMR group (2.0 mm, P＜0.001). Additionally, the ESD group had significantly more lesions with pathological grade G2[23.8%(20/84) vs. 1.3%(2/151), P＜0.001], infiltration depth reaching the submucosal layer [78.6%(66/84) vs. 51.0%(77/151), P＜0.001], and more T2 stage compared to the EMR group[15.5%(13/84) vs. 0.7%(1/151), P＜0.001]. After PSM, 49 pairs of lesions were successfully matched between the two groups. Following PSM, there were no significant differences in the en bloc resection rate [100.0%(49/49) vs. 100.0%(49/49)], complete resection rate [93.9%(46/49) vs. 100.0%(49/49)], and complication rate [0(0/49) vs. 4.1%(2/49)] between the two groups. During the follow-up period, no recurrence or distant metastasis was observed in any of the lesions in both groups. Conclusions:The combination of endoscopy and serology diagnostic strategy has the potential to enhance the accuracy of diagnosing G1 and G2 stage G-NENs and their background mucosa. Endoscopic resection surgery (EMR, ESD) is a proven and safe treatment approach for G1 and G2 stage G-NENs.

Objectives:To develop and validate predictive models for esophageal squamous cell carcinoma (ESCC) using circulating cell-free DNA (cfDNA) terminal motif analysis. The goal was to improve the non-invasive detection of early-stage ESCC and its precancerous lesions.Methods:Between August 2021 and November 2022, we prospectively collected plasma samples from 448 individuals at the Department of Endoscopy, Cancer Hospital, Chinese Academy of Medical Sciences for cfDNA extraction, library construction, and sequencing. We analyzed 201 cases of ESCC, 46 high-grade intraepithelial neoplasia (HGIN), 46 low-grade intraepithelial neoplasia (LGIN), 176 benign esophageal lesions, and 29 healthy controls. Participants, including ESCC patients and control subjects, were randomly assigned to a training set ( n=284) and a validation set ( n=122). The training cohort underwent z-score normalization of cfDNA terminal motif matrices and a selection of distinctive features differentiated ESCC cases from controls. The random forest classifier, Motif-1 (M1), was then developed through principal component analysis, ten-fold cross-validation, and recursive feature elimination. M1's efficacy was then validated in the validation and precancerous lesion sets. Subsequently, individuals with precancerous lesions were included in the dataset and participants were randomly allocated to newly formed training ( n=243), validation ( n=105), and test ( n=150) cohorts. Using the same procedure as M1, we trained the Motif-2 (M2) random forest model with the training cohort. The M2 model's accuracy was then confirmed in the validation cohort to establish the optimal threshold and further tested by performing validation in the test cohort. Results:We developed two cfDNA terminal motif-based predictive models for ESCC and associated precancerous conditions. The first model, M1, achieved a sensitivity of 90.0%, a specificity of 77.4%, and an area under the curve (AUC) of 0.884 in the validation cohort. For LGIN, HGIN, and T1aN0 stage ESCC, M1's sensitivities were 76.1%, 80.4%, and 91.2% respectively. Notably, the sensitivity for jointly predicting HGIN and T1aN0 ESCC reached 85.0%. Both the predictive accuracy and sensitivity increased in line with the cancer's progression ( P＜0.001). The second model, M2, exhibited a sensitivity of 87.5%, a specificity of 77.4%, and an AUC of 0.857 in the test cohort. M2's sensitivities for detecting precancerous lesions and ESCC were 80.0% and 89.7%, respectively, and it showed a combined sensitivity of 89.4% for HGIN and T1aN0 stage ESCC. Conclusions:Two predictive models based on cfDNA terminal motif analysis for ESCC and its precancerous lesions are developed. They both show high sensitivity and specificity in identifying ESCC and its precancerous stages, indicating its potential for early ESCC detection.