Objective: To evaluate repeated testing on blood screening platforms in confirmation of non-discriminatory reactive (NDR) samples in nucleic acid testing (NAT). Methods: A total of 102 HBsAg-negative/NAT NDR samples were collected from voluntary blood donors at Dalian Blood Center between January 2021 and December 2023. Repeated testing was performed using two NAT platforms (Cobas s201 and Panther). For the first round of repeated testing, all samples were tested 12 times on each system; for the second round, the samples which were non-reactive or only reactive once in the first round were tested an additional 8 times. Anti-HBc and anti-HBs was detected using electrochemiluminescence assay (ECA). Meanwhile, blood donors were followed up. Results: The proportion of anti-HBc+ in 102 NDR samples was 88.2%. Forty-one samples (40.2%, 41/102) and 7 samples were confirmed HBV DNA+ in first-round and second-round repeated testing, respectively. The cumulative confirmation rate of HBV DNA+ was 47.1% (48/102) after repeated testing. Extra five blood donors detected HBV DNA+ in follow-up were identified as anti-HBc+ occult hepatitis B virus infection (OBI), while no window period infection was observed. Ultimately, there were 53 HBV infected donors confirmed, 46 HBV infection-unconfirmed, and 3 HBV uninfected. No significant difference was observed between the confirmation rate of the first-round testing and the cumulative confirmation rate after the second-round testing (P>0.05). The proportion of anti-HBc+ donors was quite high in both HBV infection-confirmed (98.1%) and unconfirmed group (82.6%), and donors with seronegative and anti-HBs-only occupied a high proportion in the latter (P<0.05). Conclusion: Numerous repeated testing of NDR samples using NAT platforms cannot achieve complete confirmation of HBV infection. Supplementary anti-HBc testing can minimize potential OBI risk among NDR donors, and is low-cost and efficient.

ObjectiveTo investigate the epidemiological characteristics of human brucellosis (hereinafter referred to as brucellosis) in Baoshan City, Yunnan Province, and to provide scientific evidence for adjusting prevention and control strategies. MethodsBased on the surveillance data of reported brucellosis cases in Baoshan City from 2020 to 2023 and the information collected through individual epidemiological questionnaire surveys, the epidemic status and clinical characteristics of brucellosis in Baoshan City were analyzed using descriptive epidemiological methods. ResultsA total of 85 brucellosis cases were reported in Baoshan City from 2020 to 2023, and detailed individual information was obtained for 83 of them. Brucellosis in Baoshan City showed a clear seasonal pattern, with peak incidence from May to September. The average annual incidence rate was 0.80/100 000, with a male-to-female ratio of 7.5∶1. And 82.35% of the cases aged 30 to 60 years, with farmers being the predominant affected group. The main clinical manifestations of the cases were myalgia and arthralgia. Regarding transmission routes, 87.95% of the cases had a contact history with cattle, with livestock rearing and grazing being the main exposure modes. Most infections occurred at home. ConclusionFrom 2020 to 2023, the incidence of brucellosis in Baoshan City exhibited a fluctuating upward trend, with a peak period from May to September. Males and farmers were identified as the primary affected populations. It is recommended to strengthen livestock surveillance and control, and to enhance both awareness and self-protection capacity among high-risk groups.

ObjectiveTo investigate the distribution characteristics and correlation of paired oral and esophageal microbiota in patients with esophageal squamous cell carcinoma （ESCC）. MethodsA total of 46 pathologically confirmed ESCC patients were enrolled， and oral swabs and cancerous tissue samples were collected. The microbial composition was analyzed using 16S rRNA gene sequencing， and bioinformatics methods were employed to compare the microbiota diversity， compositional differences， and correlations between the two sites. ResultsThe Alpha diversity of the oral microbiota in ESCC patients was significantly higher than that of their esophageal microbiota， though the overall community structure was similar between the two sites. LEfSe analysis revealed that Spirochaetes， Prevotella， Catonella， Enterococcus， Fusobacterium periodonticum， Veillonella parvula， Capnocytophaga sputigena， and Neisseria subflava were significantly enriched in ESCC tumor tissues. In contrast， Actinobacteria， Proteobacteria， Streptococcus， Neisseria， Haemophilus， Streptococcus mitis， Haemophilus parainfluenzae， Porphyromonas gingivalis， Haemophilus haemolyticus， and Prevotella tannerae were significantly enriched in the oral cavity. Correlation analysis demonstrated significant associations between the two sites at taxonomic levels such as Spirochaetes， Treponema， Granulicatella elegans， Neisseria macacae， Treponema amylovorum， Fusobacterium periodonticum， and Neisseria meningitidis （P<0.05）. ConclusionThe oral and esophageal microbiota in ESCC patients exhibit both distinct differences and similarities. Oral microbiota may influence the esophageal microenvironment through migration or local immune modulation.

ObjectiveTo investigate the effect of Hezhongxiaopi granules （HZXPG） on methylnitronitrosoguanidine （MNNG）-mediated gastric mucosal cell apoptosis and the regulatory effects on the Nicalin （Ncln）-C/EBP homologous protein （CHOP） axis and endoplasmic reticulum stress （ERS）. MethodsThe cell counting kit-8 （CCK-8） assay was employed to examine the inhibitory effect of MNNG on the proliferation of GES-1 cells， and the optimal effective dose of MNNG was selected. Human gastric mucosal GES-1 cells were allocated into the control， model （MNNG， 1.562 5 mmol·L^-1）， and low/high-dose HZXPG （5% and 10% HZXPG-containing serum） groups. Phalloidin staining was employed to detect MNNG-induced morphological changes in human gastric mucosal cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was adopted to assess cell apoptosis. Annexin Ⅴ/propidium iodide （Annexin Ⅴ/PI） flow cytometry was utilized to measure the alterations in the apoptotic cycle of human gastric mucosal cells. Western blot was performed to determine the expression levels of proteins related to apoptosis， ERS， and the Ncln/CHOP axis. ResultsCompared with the control group， the model group exhibited a reduction in the surface area of GES-1 cells， endoplasmic reticulum （ER） dilation， increased early and late apoptosis rates of gastric mucosal cells， upregulated protein levels of truncated Bid （tBid）， Bcl-2-associated X protein （Bax）， cleaved-cysteinyl aspartate-specific proteinase （Caspase）-3， and cleaved-Caspase-8， and downregulated protein level of B-cell lymphoma-2 （Bcl-2） （P<0.01）. In addition， the expression levels of binding immunoglobulin protein （BiP）， Ncln， and CHOP were upregulated （P<0.01）， with enhanced Ncln expression in the ER and elevated CHOP expression in the nucleus. Compared with the model group， the high-dose HZXPG group showed a significant increase in the surface area of gastric mucosal cells. Both dose groups exhibited inhibited ER dilation and reduced early and late apoptosis rates of gastric mucosal cells （P<0.01）. Low-dose HZXPG decreased the expression of tBid and Bax （P<0.05， P<0.01） and promoted Bcl-2 expression （P<0.01）. High-dose HZXPG reduced the expression levels of tBid， Bax， cleaved-Caspase-3， and cleaved-Caspase-8 （P<0.05， P<0.01） and increased the Bcl-2 expression level （P<0.05）. Furthermore， high-dose HZXPG downregulated the expression levels of BiP， Ncln， and CHOP （P<0.05， P<0.01）， while decreasing the nuclear translocation of CHOP. ConclusionHZXPG alleviates MNNG-induced ERR-mediated apoptosis of gastric mucosal cells by inhibiting the Ncln-CHOP axis， thereby relieving gastric mucosal cell damage caused by precancerous gastric lesions.

BACKGROUND:Needle-knife release of the ligamentum flavum can effectively improve symptoms in patients with lumbar degeneration,and ultrasound guidance can increase the precision of needle-knife release;however,the specific effects of needle-knife release of the ligamentum flavum on the degenerated intervertebral discs and the possible mechanisms remain to be clarified. OBJECTIVE:To investigate the effect of ultrasound-guided needle-knife release of the ligamentum flavum. METHODS:Twenty-four New Zealand rabbits were randomized into control(n=6)and model(n=18)groups.A rabbit model of lumbar disc degeneration model was established in the model group by cutting the supraspinous and interspinous ligaments of the L5/6 and L6/7 segments to maintain a standing posture and apply axial load to the lumbar spine.After successful modeling,the model rabbits were subdivided into a control group,a model group,an ultrasonic needle-knife group,and a sham needle-knife group according to a random number table method,with six animals in each group.The ultrasonic needle-knife group underwent ultrasound-guided needle-knife release of the right yellow ligament of L7/S1,once every week,for a total of four times.The needle-knife approach in the sham needle-knife group was the same as that in the ultrasound needle-knife group,but the ligamentum flavum was not released.At 30 days after the intervention,MRI was used to observe the changes in the signal intensity of the nucleus pulposus within the L7/S1 segment.Hematoxylin-eosin staining was used to observe the morphological changes of the L7/S1 segment.Immunohistochemical staining was used to detect the expression of type I and II collagen in the nucleus pulposus of the L7/S1 segment.RT-PCR and western blot were used to detect the expression of integrin α5 and β1,p38,and nuclear factor κB in the L7/S1 segment. RESULTS AND CONCLUSION:MRI findings indicated that the nucleus pulposus of the intervertebral disc of rabbits in the model group was gray-black in color,and the gray value of the nucleus pulposus was significantly lower than that of the control group(P<0.01).The brightness of the nucleus pulposus of the intervertebral disc of the rabbits in the ultrasonic needle-knife group was elevated compared with that of the model group,and the gray value of the nucleus pulposus was higher than that of the model group(P<0.01).Results from hematoxylin-eosin staining showed that in the model group,the shape of the nucleus pulposus was irregular,the number of nucleus pulposus cells was reduced,the extracellular matrix was compressed,the fibrous ring was ruptured,the structure and boundary of the end plate were unclear,and the chondrocytes were arranged disorderly.Compared with the model group,the ultrasonic needle-knife group showed an increase in the number of the nucleus pulposus,an improvement in the rupture of the fibrous ring,and more regular arrangement of cartilage endplate cells.Results from immunohistochemical staining showed an increase in positive expression of type I collagen(P<0.01)and a decrease in positive expression of type II collagen in the nucleus pulposus of the model group compared with the control group as well as a decrease in positive expression of type I collagen and an increase in positive expression of type II collagen in the nucleus pulposus of the ultrasonic needle-knife group compared with the model group(P<0.01).RT-PCR and western blot assays showed that the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the model group were increased compared with that in the control group(P<0.01);the mRNA and protein expression of integrin α5,integrin β1,p38,and nuclear factor κB in the intervertebral discs of rabbits in the ultrasonic needle-knife group was decreased compared with that in the model group(P<0.01).To conclude,ultrasound-guided needle-knife release of the ligamentum flavum can improve the degree of lumbar disc degeneration in rabbits,which may be related to the inhibition of p38 and nuclear factor-κB expression by modulating integrin α5 and β1 expression.

Based on the traditional Chinese medicine theory that "all pain， itching， and sores are related to the heart"， this paper proposes treating recurrent aphthous ulcers from the perspective of the heart. It suggests that excessive heart fire and tissue erosion due to flaming fire in the heart meridian constitute the core pathogenesis of this condition. Hyperactive heart fire is identified as the key pathogenic factor， while heart yin deficiency， obstruction of the heart collaterals， and malnourishment of the heart spirit are considered significant contributing factors. Clinically， the treatment follows the principle of clearing heart fire as the main strategy， supplemented by nourishing yin， activating collaterals， and calming the spirit. The self-formulated Qingxin Yuchuang Formulation （清心愈疮方） serves as the base prescription， with flexible modifications incorporating the Yuyin Formulation （育阴方）， Huoxue Formulation （活血方）， and Yu'an Formulation （郁安方） to address specific syndromes involving heart yin deficiency， collateral blockage， and emotional disturbance.

Abstract Esophageal cancer is one of the most common malignant tumors of the digestive system in China , with e-sophageal squamous cell carcinoma (ESCC) being the predominant pathological type. The exact etiology of ESCC remains incompletely understood. With advances in 16S rRNA gene sequencing and metagenomics , microbial dys- biosis has been suggested to play a significant role in the pathogenesis of ESCC. Currently , research on the rela- tionship between microorganisms and ESCC is still in its early stages. This review summarizes the association be- tween oral and esophageal microbiota and ESCC , factors influencing microbial composition , and microbial commu- nities linked to ESCC prognosis , which may contribute to early detection and optimized treatment strategies for ES- CC.

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder in women of childbearing age, which can cause metabolic disorders, cardiovascular disease, ovarian cancer, uterine cancer and other complications, seriously endangering the health of the body. China has become one of the countries with the fastest increasing prevalence of PCOS, but its complex pathogenesis leads to highly heterogeneous clinical manifestations, making it difficult to completely cure. Therefore, clarifying the potential pathogenesis of PCOS is of great significance for early clinical screening, diagnosis, treatment, and prognosis. Recent studies have shown that long noncoding RNA (lncRNA) plays a dual role in the pathogenesis of PCOS and is a potential novel biomarker and intervention target. The characteristics of multi-component, multi-target, and multi-pathway action in Traditional Chinese Medicine (TCM) are consistent with the biological properties of lncRNA, which have diverse types, dual roles, and diverse locations. However, research on lncRNA mediated PCOS and how TCM can improve PCOS by regulating lncRNA is relatively scattered, which is not conducive to the recognition of its clinical value. Therefore, this article provides a systematic review of the dual regulatory mechanism, clinical value, and TCM intervention research of lncRNA in the occurrence and development of PCOS, aiming to clarify how lncRNA affects the occurrence and development of PCOS and potential treatment strategies, in order to provide new ideas for the clinical prevention and treatment of PCOS.

This article summarized Professor Liu Qiquan's clinical experience in the treatment of acne from the perspective of"internal retention and stagnation of heat pathogen".Professor Liu believes that the core pathogenesis of acne lies in the internal retention and stagnation of heat pathogen,which manifests as external skin inflammation,with the primary focus being on the heat pathogen.With the pathogen inside the body,the progress of disease and syndrome is related to"toxin,depression,blood stasis and deficiency".At the same time,the disease is also closely related to the dysfunction of the five-zang organs.Professor Liu points out that"to resist the outside,one must first settle the inside",and treats acne from the five internal organs.Based on the idea of"internal retention and stagnation of heat pathogen",according to the symptoms of the patients,the comprehensive use of methods to guide the treatment,such as penetrating turbidity,removing pathogenic factors and detoxifying heat,promoting qi and dispersing stagnant heat,cooling collaterals to control blood and breaking stasis heat,and restraining qi,softening yin and supporting deficiency heat,has been proved to have a good clinical effect.One medical case was attached as evidence.

Objective:To develop and validate a nomogram prediction model for major adverse cardiovascular events (MACE) in elderly patients with chronic heart failure (CHF) based on the ratio of homocysteine to free triiodothyronine (HCY/FT3).Methods:This research was a prognostic study. A total of 1 301 elderly patients with CHF admitted to the Department of General Practice of Shanghai Fifth People′s Hospital Affiliated to Fudan University from January 2018 to January 2023 were enrolled and divided into MACE group ( n=564) and non-MACE group ( n=737) according to whether MACE occurred at the end of the follow-up. Baseline clinical data was collected. The follow-up period was 18 months, with the follow-up deadline being June 30, 2024. The primary endpoint was the occurrence of MACE, including death from any cause or re-hospitalization due to heart failure. Multivariate logistic regression analysis was used to determine the independent risk factors for MACE of elderly patients with CHF ( P<0.05). All patients included were randomly allocated into a training cohort ( n=913) and a validation cohort ( n=388) in a 7∶3 ratio. A nomogram prediction model was developed. The model was internally validated by bootstrapping. The discriminative ability of the model was assessed by calibration curves and receiver operating characteristic (ROC) curves. Results:Multivariable logistic regression analysis demonstrated that atrial fibrillation history, lower diastolic blood pressure, elevated uric acid, elevated free thyroxine (FT4), higher HCY/FT3 ratio, and lower left ventricular ejection fraction (LVEF) were independent predictors of MACE in elderly CHF patients ( P<0.05). A nomogram model incorporating these factors was developed. Internal validation using bootstrapping showed good calibration, as both training and validation cohort calibration curves closely approximated the ideal line. ROC curve analysis indicated an area under the curve of 0.721 (95% CI: 0.661-0.782) for the nomogram in predicting MACE. Conclusions:We developed and internally validated a nomogram incorporating the HCY/FT3 ratio to predict MACE risk in elderly CHF patients. This model demonstrated acceptable discrimination and good calibration, suggesting potential clinical utility for risk stratification.