1.Comparative analyses of the detection performance of five multiplex polymerase chain reaction nucleic acid detection kits for respiratory pathogens
Fang YUAN ; Lei BI ; Jiajing LIU ; Huanru WANG ; Jun FENG ; Yuan ZHUANG ; Min CHEN ; Zheng TENG
Shanghai Journal of Preventive Medicine 2026;38(2):165-169
ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.
2.The Regulatory Effects and Mechanisms of Piezo1 Channel on Chondrocytes and Bone Metabolic Dysregulation in Osteoarthritis
Yan LI ; Tao LIU ; Yu-Biao GU ; Hui-Qing TIAN ; Lei ZHANG ; Bi-Hui BAI ; Zhi-Jun HE ; Wen CHEN ; Jin-Peng LI ; Fei LI
Progress in Biochemistry and Biophysics 2026;53(3):564-576
Osteoarthritis (OA), a highly prevalent degenerative joint disease worldwide, is defined by articular cartilage degradation, abnormal bone remodeling, and persistent chronic inflammation. It severely compromises patients’ quality of life, and currently, there is no radical cure. Abnormal mechanical stress is widely regarded as a core driver of OA pathogenesis, and the exploration of mechanical signal perception and transduction mechanisms has become crucial for deciphering OA’s pathophysiological processes. Piezo1, a key mechanosensitive cation channel belonging to the Piezo protein family, has recently gained significant attention due to its pivotal role in mediating cellular responses to mechanical stimuli in joint tissues. This review systematically examines Piezo1’s expression patterns, regulatory mechanisms, and pathological functions in OA, with a particular focus on its dual roles in modulating chondrocyte homeostasis and bone metabolism disorders, while also delving into the underlying molecular signaling pathways and potential therapeutic implications. Piezo1, consisting of approximately 2 500 amino acids and forming a unique trimeric propeller-like structure, is widely expressed in chondrocytes, osteocytes, mesenchymal stem cells, and synovial cells. It exhibits permeability to cations such as Ca2+, K+, and Na+, and directly responds to membrane tension changes induced by mechanical stimuli like fluid shear stress and mechanical overload. In OA patients and animal models, Piezo1 expression is significantly upregulated, especially in cartilage regions subjected to abnormal mechanical stress (e.g., human temporomandibular joint cartilage). This overexpression is closely associated with aggravated cartilage degeneration, increased chondrocyte apoptosis, accelerated cellular senescence, and intensified inflammatory responses. Mechanical overload and pro-inflammatory cytokines (e.g., IL-1β) are key inducers of Piezo1 upregulation: IL-1β activates the PI3K/AKT/mTOR signaling pathway to enhance Piezo1 expression, forming a pathogenic positive feedback loop that inhibits chondrocyte autophagy, promotes apoptosis, and further accelerates joint degeneration. Mechanistically, Piezo1 mediates OA progression through multiple interconnected pathways. When activated by mechanical stress, Piezo1 triggers excessive Ca2+ influx, leading to endoplasmic reticulum stress (ERS) and mitochondrial dysfunction, which directly induce chondrocyte apoptosis. This process involves the activation of downstream signaling cascades such as cGAS-STING and YAP-MMP13/ADAMTS5. YAP, a transcriptional regulator, upregulates the expression of matrix metalloproteinase 13 (MMP13) and aggrecanase (ADAMTS5), thereby accelerating cartilage matrix degradation. Additionally, Piezo1-driven Ca2+ overload promotes the accumulation of reactive oxygen species (ROS) and upregulates senescence markers (p16 and p21), accelerating chondrocyte senescence via the p38MAPK and NF-κB pathways. Senescent chondrocytes secrete senescence-associated secretory phenotype (SASP) factors (e.g., IL-6, IL-1β), further amplifying joint inflammation. In terms of bone metabolism, Piezo1 maintains joint homeostasis by promoting the differentiation of fibrocartilage stem cells into chondrocytes and balancing bone formation and resorption through regulating the FoxC1/YAP axis and RANKL/OPG ratio. Therapeutically, targeting Piezo1 shows promising potential. Preclinical studies have demonstrated that Piezo1 inhibitors (e.g., GsMTx4) can reduce joint damage and alleviate pain in OA mice. Simultaneously, siRNA-mediated co-silencing of Piezo1 and TRPV4 (another mechanosensitive channel) decreases intracellular Ca2+ concentration, inhibits chondrocyte apoptosis, and promotes cartilage repair. Conditional knockout of Piezo1 using Gdf5-Cre transgenic mice alleviates cartilage degeneration in post-traumatic OA models by downregulating MMP13 and ADAMTS5 expression. Despite existing challenges, such as off-target effects of inhibitors, inefficient local drug delivery, and interindividual genetic variability, strategies like developing selective Piezo1 antagonists, optimizing targeted nanocarriers, and combining Piezo1-targeted therapy with physical therapy provide viable avenues for clinical translation. The authors propose that Piezo1 serves as a critical therapeutic target for OA, and future research should focus on deciphering its context-dependent regulatory networks, developing tissue-specific intervention strategies, and validating their efficacy and safety in clinical trials to address the unmet medical needs of OA patients.
3.Clinical characteristics of patients with brucellosis complicated with epididymo-orchitis
Lei ZOU ; Yao CHENG ; Qingfeng GAO ; Luo SUN ; Ci WANG ; Shuning SUI ; Jingyao LIU ; Baiqiang ZHANG ; Huimei BI
Chinese Journal of Endemiology 2025;44(10):836-840
Objective:To explore the clinical characteristics of patients with brucellosis complicated with epididymo-orchitis (Brucellar epididymo-orchitis, BEO), so as to provide reference for clinical diagnosis and treatment of BEO.Methods:General and clinical data of 293 male patients with acute brucellosis admitted to the Beidahuang Industry Group General Hospital from January 2023 to December 2024 were retrospectively collected. They were divided into a BEO group (30 cases) and a non-BEO group (263 cases) based on the presence or absence of epididymo-orchitis. Epidemic characteristics, clinical manifestations and laboratory examination results were compared and analyzed.Results:Among 293 male patients with acute brucellosis, the incidence of BEO was 10.24% (30/293). Their age was mainly concentrated in 45 - < 60 years old (53.33%, 16/30), and occupation was mostly farmers (63.33%, 19/30). There were no statistically significant differences in the distribution of age, occupation, exposure history and onset season between the BEO group and the non-BEO group ( P > 0.05). The distribution of abdominal pain and urinary frequency/urgency/pain symptoms was compared, and the differences were statistically significant ( P < 0.05). White blood cell count (WBC), neutrophil count (NEUT), and C-reactive protein (CRP) levels in the BEO group were higher than those in the non-BEO group ( P < 0.05), while the levels of cytokines interleukin (IL)-6 and interferon-γ (IFN-γ) were lower than those in the non-BEO group ( P < 0.05). After 6 - 8 weeks of hospitalization, the levels of WBC, NEUT, CRP, albumin, alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transpeptidase, alkaline phosphatase, α-hydroxybutyrate dehydrogenase, lactate dehydrogenase, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor-α, and IFN-γ in patients of the BEO group were significantly different from before treatment ( P < 0.05). Twenty-eight patients were cured, 1 patient underwent orchiectomy, and 1 patient experienced recurrence. Conclusions:Middle-aged male patients with brucellosis are more prone to BEO. Clinical manifestations and laboratory tests have certain diagnostic value for BEO. Suspected BEO patients should be diagnosed and treated as early as possible to reduce the occurrence of adverse prognosis.
4.Clinical study on peripheral blood 25-hydroxy vitamin D, TBNK lymphocyte subsets, and cytokines levels in patients with brucellosis
Ci WANG ; Baiqiang ZHANG ; Qingfeng GAO ; Xun ZHOU ; Kun ZHOU ; Yanli LI ; Shuning SUI ; Lei ZOU ; Huimei BI
Chinese Journal of Endemiology 2025;44(10):786-791
Objective:To learn about the levels of 25-hydroxy vitamin D (25-OH VD), TBNK lymphocyte subsets, and cytokines in peripheral blood of patients with brucellosis.Methods:A prospective design was adopted, one hundred patients with brucellosis admitted to the Department of Infectious Diseases, Beidahuang Industry Group General Hospital from May 2024 to February 2025 were selected as the brucellosis group, and one hundred healthy individuals who underwent physical examinations at the hospital during the same period were selected as the control group. The peripheral blood 25-OH VD levels were detected by chemiluminescence method. Further, 100 patients with brucellosis were divided into a brucellosis combined with osteoarthritis group (74 cases) and a brucellosis without osteoarthritis group (26 cases). Flow cytometry was used to detect the counts of peripheral blood TBNK lymphocyte subsets and cytokine levels. Meanwhile, Spearman rank correlation was used to analyze the correlation between peripheral blood 25-OH VD levels and TBNK lymphocyte subsets counts as well as cytokine levels in patients with brucellosis complicated by osteoarthritis.Results:The peripheral blood 25-OH VD level in the brucellosis group [20.31 (15.74, 24.35) ng/ml] was significantly lower than that of the control group [25.18 (21.13, 29.59) ng/ml], and the difference was statistically significant ( Z = - 5.07, P < 0.001). The peripheral blood 25-OH VD level [18.05 (13.79, 23.74) vs 22.43 (19.93, 28.25) ng/ml], CD4 + T cell count [(860 ± 275) vs (1 036 ± 376) cells/μl], and interleukin (IL)-6 levels [4.17 (2.14, 9.41) vs 7.83 (5.97, 11.34) ng/L] in the brucellosis combined with osteoarthritis group were significantly lower than those in the brucellosis without osteoarthritis group ( Z/t = - 2.88, 2.20, - 2.85, P = 0.004, 0.035, 0.004). Correlation analysis showed that the peripheral blood 25-OH VD level in patients with brucellosis complicated by osteoarthritis was positively correlated with the counts of CD45 +, CD3 + T, CD4 + T, CD8 + T, and natural killer cells ( r = 0.31, 0.26, 0.25, 0.25, 0.25, P = 0.007, 0.027, 0.032, 0.031, 0.032), and negatively correlated with IL-17A level ( r = - 0.40, P < 0.001). Conclusion:Patients with brucellosis have insufficient 25-OH VD, and those with osteoarthritis have lower 25-OH VD level, CD4 + T cell count, and IL-6 level than those without osteoarthritis.
5.Comparative efficacy of internal fixation with video thoracoscopy-assisted rib plating and open thoracotomy in the treatment of multiple rib fracture
Lei BI ; Zhengyu CHEN ; Yiping DENG ; Cheng AI ; Fuyu YANG ; Zhongzhu LYU
Chinese Journal of Trauma 2025;41(3):289-296
Objective:To compare the efficacy of internal fixation with video thoracoscopy-assisted rib plating and open thoracotomy in the treatment of multiple rib fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 65 patients with multiple rib fracture who were admitted to Affiliated Bishan Hospital of Chongqing Medical University between May 2021 and May 2023, including 42 males and 23 females, aged 19-75 years [(51.6±7.0)years]. Of all, 33 patients were treated with internal fixation with video thoracoscopy-assisted rib plating (thoracoscopy group), while other 32 patients treated with internal fixation with open thoracotomy (thoracotomy group). Two groups were compared in terms of surgical incision length, intraoperative blood loss, surgical duration, duration of postoperative drainage tube placement, postoperative chest tube drainage, and length of hospital stay. Postoperative pain was assessed using the visual analogue scale (VAS) at 6, 12, 24, 48, and 72 hours postoperatively. Forced vital capacity (FVC), forced expiratory volume in the first second (FEV1), and peak expiratory flow (PEF) were detected preoperatively, at 7 days, 6 months postoperatively and at the last follow-up. The excellent and good rate of fracture healing was evaluated at 6 months postoperatively and at the last follow-up. The incidence of postoperative complications was also assessed.Results:All the patients were followed up for 12-24 months [(15.2±2.2)months]. The surgical incision length, intraoperative blood loss, and surgical duration were (4.3±1.5)cm, (65.2±15.0)ml, and (68.8±13.1)minutes in the thoracoscopy group, shorter or less than (7.2±1.7)cm, (93.3±16.3)ml, and (93.7±15.9)minutes in the thoracotomy group ( P<0.01). The duration of drainage tube placement, postoperative chest tube drainage volume and length of hospital stay were (3.8±1.5)days, (357.3±38.6)ml and (12.3±1.7)days in the thoracoscopy group, shorter or less than (5.9±1.8)days, (424.9±45.4)ml, and (18.6±2.5)days in the thoracotomy group ( P<0.01). At 6, 12, 24, and 48 hours postoperatively, the VAS scores in the thoracoscopy group were (5.1±1.6)points, (4.7±1.5)points, (4.2±1.5)points, and (3.9±1.3)points, significantly lower than those in the thoracotomy group [(8.4±1.8)points, (7.3±1.5)points, (6.3±1.3)points, and (5.2±1.2)points] ( P<0.01). There was no statistically significant difference in the VAS scores between the two groups at 72 hours postoperatively ( P>0.05). There were no statistically significant differences in FVC, FEV1 and PEF between the two groups preoperatively, at 6 months postoperatively and at the last follow-up ( P>0.05). At 7 days postoperatively, FVC, FEV1 and PEF were (4.17±0.25)L, (2.24±0.24)L, and (5.53±0.50)L/s in the thoracoscopy group, significantly higher than those in the thoracotomy group [(4.01±0.23)L, (2.12±0.21)L, and (5.23±0.42)L/s] ( P<0.05). At 6 months postoperatively, the excellent and good rate was 94% (31/33) in the thoracoscopy group and 97% (31/32) in the thoracotomy group ( P>0.05). At the last follow-up, the excellent and good rate in both groups were 100% ( P>0.05). The incidence of complications was 15% (5/33) in the thoracoscopy group, lower than 41% (13/32) in the thoracotomy group ( P<0.05). Conclusion:Compared with internal fixation with open thoracotomy in the treatment of multiple rib fracture, the internal fixation with video thoracoscopy-assisted rib plating has the advantages of less surgical trauma, milder pain at the early stage after surgery, earlier postoperative recovery of pulmonary function and fewer complications.
6.Impact of Polygonum cuspidatum and polydatin on lipid deposition in adipose tissue of obese mice
Bi-lin XU ; Lu-guang SHENG ; Dan-dan LIU ; Wei-bin LIU ; Tao LEI ; Qing-guang CHEN ; Hao LU
Chinese Traditional Patent Medicine 2025;47(9):2912-2917
AIM To investigate the effects of Polygonum cuspidatum and polydatin on lipid deposition in adipose tissue of high-fat diet-induced obese mice.METHODS Forty male C57BL/6J mice were randomly assigned to either a control group(10 mice)fed standard chow or a diet-induced obesity(DIO)group(30 mice)fed a high-fat diet for 8 weeks.The successful mouse models were randomly assigned to the model group,the polydatin group(250 mg/kg)and the P.cuspidatum group(4.5 g/kg),with 8 mice in each group,to resume their high-fat diet during the following 8 weeks corresponding drug administration by gavage.Weekly body weight measurements were recorded for all mice.Serum TG,TC and LDL levels were quantified post-treatment.Histopathological assessment of adipose tissue was performed using HE staining.The mRNA expressions of AMPK,SREBP-1c and FAS in adipose tissue were analyzed by RT-qPCR.The protein expressions of p-AMPK,SREBP-1c and FAS in adipose tissue was detected by Western blot.RESULTS Compared to the control group,the model group displayed significantly higher body weight,inguinal fat weight and epididymal fat weight(P<0.05);elevated serum TG,TC and LDL levels(P<0.05);markedly enlarged volumes of inguinal and epididymal adipocytes(P<0.01);reduced p-AMPK protein expression in inguinal adipose tissue(P<0.01);and upregulated mRNA and protein expressions of SREBP-1c and FAS(P<0.05,P<0.01).Compared to the model group,both the P.cuspidatum group and polygonin group exhibited significantly reduced body weight and inguinal fat weight(P<0.05);decreased serum TG and TC levels(P<0.05);reduced inguinal adipocyte size(P<0.01);elevated p-AMPK protein expression in inguinal adipose tissue(P<0.01);and downregulated mRNA and protein expressions of SREBP-1c and FAS(P<0.05,P<0.01).CONCLUSION P.cuspidatum and polydatin significantly increases p-AMPK expression while decreasing SREBP-1c and FAS levels in adipose tissue.This regulatory effect likely contributes to reduction of body weight in obese mice through suppression of lipogenesis.
7.Phase II study of radiotherapy combined with anlotinib in the treatment of inoperable non-small cell lung cancer
Haiyuan LI ; Yupei YUAN ; Tao ZHANG ; Lei DENG ; Wenyang LIU ; Wenqing WANG ; Xin WANG ; Jima LYU ; Zongmei ZHOU ; Qinfu FENG ; Zefen XIAO ; Nan BI ; Jianyang WANG
Chinese Journal of Radiation Oncology 2025;34(4):334-339
Objective:To analyze the safety and short-term efficacy of thoracic radiotherapy combined with anlotinib in the treatment of inoperable non-small cell lung cancer (NSCLC).Methods:A prospective study was conducted on patients with unresectable locally advanced NSCLC who were intolerant to concurrent chemoradiotherapy and treated at the Department of Radiation Oncology, Cancer Hospital, Chinese Academy of Medical Sciences, from October 2020 to September 2023. Anlotinib was administered orally concurrently with radiotherapy (days 1-14, 21 days per cycle, for 3 cycles). Adverse effects and short-term tumor recurrence were observed from the beginning of radiotherapy to the 3-month post-radiotherapy. Kaplan-Meier method was used to calculate progression-free survival (PFS) and overall survival (OS) rates from the date of initial treatment (induction therapy), and intergroup comparisons were performed using the log-rank test.Results:The median age was 62 years (range:42-76 years), with a male predominance ( n=36, 88%) of the included 41 patients. The incidence of grade 3-4 acute hematologic adverse events was 20% (8 cases); the incidence of grade 3 hemoptysis was 2% (1 case), with no grade 4 hemoptysis; the incidence of grade 3-4 radiation pneumonitis was 10% (4 cases). No grade 5 adverse events were observed in the entire cohort. With a median follow-up of 19.7 months (range: 7.1-50.1 months), 19 patients (46%) experienced recurrence, including 4 patients (10%) with local recurrence, 6 patients (15%) with regional lymph node recurrence, and 11 patients (27%) with distant metastases. The 1-year PFS rate was 78.3%. 8 patients (20%) died, including 3 patients died from COVID-19 infection during the follow-up period, 1 patient who died from hypostatic pneumonia due to prolonged bed rest after cerebral infarction, and 4 patients died from tumor-related causes. The 1-year OS rate was 78.0%. Conclusions:Thoracic radiotherapy combined with anlotinib demonstrates good safety, manageable adverse events, and favorable short-term efficacy in NSCNC patients intolerant to concurrent chemoradiotherapy.
8.Prognostic value of single PET-CT after chemotherapy combined with immunotherapy in patients with non-small cell lung cancer treated with radiotherapy
Zhenghui MA ; Yuqi WU ; Guangqian JI ; Zongmei ZHOU ; Qinfu FENG ; Zefen XIAO ; Jima LYU ; Xin WANG ; Jianyang WANG ; Wenyang LIU ; Lei DENG ; Wenqing WANG ; Nan BI ; Junlin YI ; Tao ZHANG
Chinese Journal of Radiation Oncology 2025;34(11):1111-1116
Objective:To evaluate the role of a single PET-CT scan in predicting survival and prognosis in patients with non-small cell lung cancer (NSCLC) who did not undergo surgery but received radiotherapy after neoadjuvant chemotherapy combined with immunotherapy.Methods:A retrospective analysis was conducted on the data of 23 NSCLC patients treated at the Cancer Hospital of the Chinese Academy of Medical Sciences from May 2022 to June 2024. All patients were pathologically confirmed, received neoadjuvant chemotherapy combined with immunotherapy, did not undergo surgery for various reasons, and instead received radiotherapy. Each patient underwent only one PET-CT scan after neoadjuvant chemotherapy combined with immunotherapy and before radiotherapy. According to the maximum standardized uptake value (SUV max) on PET-CT, patients were divided into the low-uptake group (SUV max < 8, n=12) and high-uptake group (SUV max ≥ 8, n=11). Survival analysis was performed using the Kaplan-Meier method with survival curves plotted. Univariate analysis of influencing factors of survival was conducted using the Cox proportional hazards regression model. Clinical characteristics and survival outcomes of the two groups were compared, including progression-free survival (PFS) and overall survival (OS). Results:The 1-year PFS rates were 100% in the low-uptake group, 54.5% in the high-uptake group. This difference was statistically significant ( P=0.007). The 1-year and 2-year OS rates were both 100% in the low-uptake group, the 1-year and 2-year OS rates were both 90.9% in the high-uptake group, with no statistically significant difference ( P=0.394). Univariate Cox analysis identified age as an independent factor affecting PFS. Conclusions:For NSCLC patients who did not undergo surgical resection but received radiotherapy after neoadjuvant chemotherapy combined with immunotherapy, a single PET-CT scan before radiotherapy has potential value in predicting PFS. However, clinical studies with larger sample size and longer follow-up are required to evaluate its predictive value for OS.
9.Systemic inflammatory score predicts survival of patient with unresectable stage Ⅲ non-small cell lung cancer treated by definitive chemoradiotherapy combined with consolidation immunotherapy
Shihong LUO ; Yupei YUAN ; Yu WANG ; Yin YANG ; Tao ZHANG ; Lei DENG ; Wenyang LIU ; Wenqing WANG ; Xin WANG ; Jima LYU ; Zongmei ZHOU ; Jianyang WANG ; Nan BI
Chinese Journal of Radiation Oncology 2025;34(10):993-1000
Objective:To analyze the prognostic value of systemic inflammatory score (SIS) in patients with unresectable stage Ⅲ non-small cell lung cancer (NSCLC) treated by definitive chemoradiotherapy (dCRT) combined with or without consolidation immunotherapy with immune checkpoint inhibitor (ICI).Methods:The medical record data of 229 patients who received dCRT from January 2014 to December 2017 and 183 patients who received dCRT combined with any form of ICI (induction, concurrent, consolidation or combination) from August 2018 to August 2022 in the Cancer Hospital, Chinese Academy of Medical Sciences were retrospectively analyzed. Upon admission, 1 and 3 months after treatment (efficacy evaluation) and upon tumor recurrence, peripheral blood count was collected, and neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and SIS were calculated, respectively. The SIS before, 1 and 3 months after treatment was defined as SIS 0, SIS 1 and SIS 3, respectively. Overall survival (OS) was considered as the primary endpoint. All patients were divided into dCRT group and dCRT+ICI group according to whether received immunotherapy, and then divided into different subgroups based on the cutoff value of SIS determined by X-Tile software. The prognostic value of SIS was evaluated by Kaplan-Meier survival analysis. The area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the predictive efficiency. The predictive value of SIS was compared with inflammatory indexes (NLR, PLR) and independent prognostic factors. Results:In the dCRT group, the optimal cutoff value of SIS 0 was 590×10 9 and 530×10 9 in the dCRT+ICIs group. Univariate and multivariate analyses indicated that SIS 0 was an independent predictive factor of OS, progression - free survival (PFS), local - recurrence free survival (LRFS) and distant metastasis free survival (DMFS) in the dCRT group, but not associated with DMFS in the dCRT+ICI group. In the dCRT group, SIS 1>970×10 9 (optimal cutoff value) predicted poor OS ( HR=2.512, 95% CI=1.622-3.198, P<0.001), PFS ( HR=1.726, 95% CI=1.187-2.509, P=0.004), and DMFS ( HR=1.625, 95% CI=1.029-2.564, P=0.037). In the dCRT+ICI group, SIS 3>1570×10 9 (optimal cutoff value) indicated poor OS ( HR=5.107, 95% CI=1.731-15.069, P=0.003). In both groups, the AUC of SIS was higher than NLR, PLR and other traditional clinicopathological predictive indexes except T stage. Conclusions:SIS before treatment can be considered as an independent, dependable and easily acquired prognostic marker in patients with unresectable stage Ⅲ NSCLC treated by dCRT or dCRT+ICI. In the dCRT+ICI group, the optimal time point of post-radiotherapy SIS (3 months after treatment) is postponed than that (1 month after treatment) in the dCRT group.
10.Establishment and application of triple qRT-PCR for detection of hepatitis A virus, genus Paslahepevirus and genus Rocahepevirus
Jingchen BI ; Wenjiao YIN ; Lei YANG
Chinese Journal of Experimental and Clinical Virology 2025;39(1):115-121
Objective:In order to make up for the missed detection of the genus Rocahepevirus (HEV-C) infection in the current clinical test method and to quickly differentiate the infection from the hepatitis A virus (HAV) infection, a triple real-time fluorescence quantitative RT-PCR(qRT-PCR)detection method was established to simultaneously detect HAV, genus Paslahepevirus (HEV-A), and HEV-C. Methods:Three pairs of amplification primers and three TaqMan probes were selected to optimize the reaction system and amplification temperature to construct a triple qRT-PCR method ; standard curves were established by plasmid and the sensitivity of the method was evaluated; specificity was evaluated by other viruses; stability was evaluated by nucleic acid of HAV, HEV-A and HEV-C; samples from suspected hepatitis A or E cases were detected and compared with HAV (or HEV) IgM antibody and nested RT-PCR (nRT-PCR) detection result.Results:The correlation coefficient R2 of three standard curves of triple qRT-PCR were all greater than 0.99, and the slopes were close to -3.3. The minimum detection limits of HAV, HEV-A and HEV-C plasmids were 8 copies/μl, 5 copies/μl and 8 copies/μl, respectively. There was no cross reaction with hepatitis B virus and hepatitis C virus, etc. The coefficients of variation of intra-and inter-batch tests were less than 5%. The positive rates of HAV IgM antibody test, HAV nRT-PCR and triple qRT-PCR were 95.1% (58/61), 83.6% (51/61) and 83.6% (51/61) for serum from suspected hepatitis A cases, respectively. The coincidence rate of HAV IgM antibody test and HAV nRT-PCR (or triple qRT-PCR) was 85.2% (52/61), the difference was statistically significant ( χ2=4.00, P=0.039). The positive rates of HEV IgM antibody test, HEV nRT-PCR and triple qRT-PCR were 89.7%(61/68), 69.1%(47/68) and 72.1%(49/68) for serum from suspected hepatitis E cases, respectively. The coincidence rate of HEV IgM antibody test and triple qRT-PCR was 79.4% (54/68), difference was statistically significant ( χ2=8.64, P=0.002). The positive rates of HEV nRT-PCR and triple qRT-PCR for stool samples from suspected hepatitis E cases were 80.0% (20/25) and 76.0% (19/25), respectively; the coincidence rate was 96% (24/25), with no significant difference ( χ2=0, P>0.05). Conclusions:This study established a triple qRT-PCR detection method that can simultaneously and rapidly detect hepatitis A virus, genus Paslahepevirus or genus Rocahepevirus. It has high sensitivity, specificity and stability, and is suitable for rapid detection of specimens from patients recently infected with hepatitis A virus or hepatitis E virus.

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