Objective:To explore the predictive value of very low frequency (VLF) for the efficacy of vitamin D treatment in children with postural orthostatic tachycardia syndrome (POTS).Methods:A retrospective case-control study was conducted. A total of 65 POTS children from the Department of Pediatrics of the Second Hospital of Lanzhou University from November 2023 to May 2024 were selected as the research subjects, while 102 healthy children from the same period were selected as the control group. The 25 (OH) D levels between POTS children and healthy children were compared. The frequency of eight main symptoms, including palpitations, headache, cold sweat, blurred vision, chest tightness, dizziness, fainting, and digestive discomfort in the POTS group were analyzed, the symptom scores of the orthostatic intolerance were calculated, and the parameters related to short-range heart rate variability (HRV) during the upright test, head-up tilt test (HUTT) were collected. The correlation between 25 (OH) D levels and symptom scores of orthostatic intolerance symptoms and HRV parameters in children with POTS were analyzed by Pearson correlation analysis. The total symptom scores of children with POTS were compared for 3 months before treatment and 2 months after treatment with vitamin D 800 U/d. The difference in score before and after treatment was ≥2 as responsive group, and the score difference before and after treatment was <2 as non-responsive group. The demographic characteristics including age, gender, height, weight, body mass index, as well as HRV parameters such as the standard deviation of nn intervals (SDNN), the standard deviation of the average NN intervals (SDANN), the root mean square of successive differences (rMSSD), low-frequency (LF), high-frequency (HF), very-low-frequency (VLF) were compared between the responsive group and non-responsive group. Multivariate Logistic regression was analyzed to screen independent influencing factors of vitamin D treatment efficacy for POTS. The marker molecules related to the efficacy of vitamin D treatment in children with POTS were analyzed through receiver operating characteristic (ROC) curve.Results:A total of 167 children were included in this study. There were 65 children in the POTS group, 38 males and 27 females, with the age of (10.6±2.2) years. There were 102 healthy children in the control group, 53 males and 49 females, with the age of (11.2±1.8) years.There was no statistically significant difference in gender and age between the two groups (both P>0.05). The serum 25 (OH) D level in children in the POTS group was (37±18)nmol/L, lower than that in the control group (61±17)nmol/L ( P<0.05).Pearson correlation analysis showed that serum 25(OH)D was positively correlated with VLF, SDNN and HF ( r=0.43, 0.65, 0.36, respectively, all P<0.05). Among the POTS children, there were 48 cases in responsive group and 17 cases in non-responsive group. Univariate analysis revealed that responsive group exhibited significantly lower values in age, SDNN, SDANN, and VLF compared to the non-responsive group (all P<0.05). Multivariate Logistic regression analysis showed that age ( OR=1.47, 95% CI 1.08-2.01), SDNN ( OR=1.01, 95% CI 1.00-1.22) and VLF ( OR=1.01, 95% CI 1.00-1.15) were independent influencing factors in the effect of vitamin D in POTS children (all P<0.05). ROC curve analysis showed that the area under the curve of VLF predicting response to vitamin D treatment for POTS was 0.72 (95% CI 0.58-0.85, P=0.008). When the cutoff value was 1 272 ms 2, the sensitivity and specificity were both 70%. Conclusion:VLF may help predict the efficacy of vitamin D treatment in children with POTS.

Objective To analyze the expression levels of natural cytotoxicity receptor(NCR)NKp46,NKp30,NKG2A and NKG2D in peripheral blood of non-small cell lung cancer(NSCLC)patients and its clinical diagnostic value.Methods A total of 50 NSCLC patients admitted to Affiliated Shijiazhuang Ping'an Hospital of Hebei University of Chinese Medicine from October 2021 to August 2022 were selected as the study group(NSCLC group).Meanwhile,20 healthy individuals who underwent physical examinations during the same period were selected as the control group.Peripheral blood samples were collected and the positive expression rate of NKp4,NKp30,NKG2A and NKG2D on NK cell surface was detected by flow cytometry.Clinical data from patients was collected and the correlation between the positive expression rate of NKp46,NKp30,NKG2A and NKG2D and pathological type,TNM staging and survival time were analyzed by One-way ANOVA/t test.Results Compared to control group,the positive expression rate of NKp46 on NK cells in peripheral blood of NSCLC patients were significantly elevated(42.09%±18.55%vs 25.92%±14.03%),the difference was statistically significant(t=3.511,P＜0.01).Comparison of the positive expression rates of NKG2A,NKG2D and NKp30 between the two groups and the differences were not statisticany significant(t=0.447,0.536,1.941,all P＞0.05).Subgroup analysis of NK cells revealed a significant increase in the proportion of CD56brightCD16+NK subgroups in NSCLC group compared to control group(3.76%±1.91%vs 2.42%±0.85%),the difference was statistically significant(t=3.017,P＜0.01).The positive expression rate of NKp46 in the CD56brightCD16+NK subgroup and CD56dimCD16+NK subgroup significantly increased(59.64%±21.12%vs 43.91%±16.04%,51.32%±19.84%vs 38.69%±15.12%),the differences were statistically significant(t=2.999,2.562,all P＜0.01).There was no significant difference in the positive expression rate of NKp46,NKG2A,NKG2D and NKp30 among squamous cell carcinoma and adenocarcinoma in lung(t=0.188～0.600,all P＞0.05).As TNM progresses,the positive expression rate of NKp46 on peripheral blood NK cells in NSCLC patients decreases,but the trend is of no statistical difference(F=2.381,P=0.10).Compared to patients with survival period≤12 months,the positive expression rate of NKp46 in patients with survival period>12 months was significantly higher(49.77%±17.52%vs 36.71%±15.41%),the difference was statistically significant(t=2.716,P＜0.01).Conclusion The positive expression rate of NKp46 in peripheral blood NK cells of NSCLC patients is increased,mainly in the CD56dimCD16+NK subgroup and CD56brightCD16+NK subgroup.The expression of NKp46 is related to the survival time of NSCLC patients,but not to the pathological type.Peripheral blood NCR NKp46 can serve as a potential biomarker for the diagnosis and prognostic evaluation of NSCLC.

Objective To analyze the expression levels of natural cytotoxicity receptor(NCR)NKp46,NKp30,NKG2A and NKG2D in peripheral blood of non-small cell lung cancer(NSCLC)patients and its clinical diagnostic value.Methods A total of 50 NSCLC patients admitted to Affiliated Shijiazhuang Ping'an Hospital of Hebei University of Chinese Medicine from October 2021 to August 2022 were selected as the study group(NSCLC group).Meanwhile,20 healthy individuals who underwent physical examinations during the same period were selected as the control group.Peripheral blood samples were collected and the positive expression rate of NKp4,NKp30,NKG2A and NKG2D on NK cell surface was detected by flow cytometry.Clinical data from patients was collected and the correlation between the positive expression rate of NKp46,NKp30,NKG2A and NKG2D and pathological type,TNM staging and survival time were analyzed by One-way ANOVA/t test.Results Compared to control group,the positive expression rate of NKp46 on NK cells in peripheral blood of NSCLC patients were significantly elevated(42.09%±18.55%vs 25.92%±14.03%),the difference was statistically significant(t=3.511,P＜0.01).Comparison of the positive expression rates of NKG2A,NKG2D and NKp30 between the two groups and the differences were not statisticany significant(t=0.447,0.536,1.941,all P＞0.05).Subgroup analysis of NK cells revealed a significant increase in the proportion of CD56brightCD16+NK subgroups in NSCLC group compared to control group(3.76%±1.91%vs 2.42%±0.85%),the difference was statistically significant(t=3.017,P＜0.01).The positive expression rate of NKp46 in the CD56brightCD16+NK subgroup and CD56dimCD16+NK subgroup significantly increased(59.64%±21.12%vs 43.91%±16.04%,51.32%±19.84%vs 38.69%±15.12%),the differences were statistically significant(t=2.999,2.562,all P＜0.01).There was no significant difference in the positive expression rate of NKp46,NKG2A,NKG2D and NKp30 among squamous cell carcinoma and adenocarcinoma in lung(t=0.188～0.600,all P＞0.05).As TNM progresses,the positive expression rate of NKp46 on peripheral blood NK cells in NSCLC patients decreases,but the trend is of no statistical difference(F=2.381,P=0.10).Compared to patients with survival period≤12 months,the positive expression rate of NKp46 in patients with survival period>12 months was significantly higher(49.77%±17.52%vs 36.71%±15.41%),the difference was statistically significant(t=2.716,P＜0.01).Conclusion The positive expression rate of NKp46 in peripheral blood NK cells of NSCLC patients is increased,mainly in the CD56dimCD16+NK subgroup and CD56brightCD16+NK subgroup.The expression of NKp46 is related to the survival time of NSCLC patients,but not to the pathological type.Peripheral blood NCR NKp46 can serve as a potential biomarker for the diagnosis and prognostic evaluation of NSCLC.

Objective:To explore the predictive value of very low frequency (VLF) for the efficacy of vitamin D treatment in children with postural orthostatic tachycardia syndrome (POTS).Methods:A retrospective case-control study was conducted. A total of 65 POTS children from the Department of Pediatrics of the Second Hospital of Lanzhou University from November 2023 to May 2024 were selected as the research subjects, while 102 healthy children from the same period were selected as the control group. The 25 (OH) D levels between POTS children and healthy children were compared. The frequency of eight main symptoms, including palpitations, headache, cold sweat, blurred vision, chest tightness, dizziness, fainting, and digestive discomfort in the POTS group were analyzed, the symptom scores of the orthostatic intolerance were calculated, and the parameters related to short-range heart rate variability (HRV) during the upright test, head-up tilt test (HUTT) were collected. The correlation between 25 (OH) D levels and symptom scores of orthostatic intolerance symptoms and HRV parameters in children with POTS were analyzed by Pearson correlation analysis. The total symptom scores of children with POTS were compared for 3 months before treatment and 2 months after treatment with vitamin D 800 U/d. The difference in score before and after treatment was ≥2 as responsive group, and the score difference before and after treatment was <2 as non-responsive group. The demographic characteristics including age, gender, height, weight, body mass index, as well as HRV parameters such as the standard deviation of nn intervals (SDNN), the standard deviation of the average NN intervals (SDANN), the root mean square of successive differences (rMSSD), low-frequency (LF), high-frequency (HF), very-low-frequency (VLF) were compared between the responsive group and non-responsive group. Multivariate Logistic regression was analyzed to screen independent influencing factors of vitamin D treatment efficacy for POTS. The marker molecules related to the efficacy of vitamin D treatment in children with POTS were analyzed through receiver operating characteristic (ROC) curve.Results:A total of 167 children were included in this study. There were 65 children in the POTS group, 38 males and 27 females, with the age of (10.6±2.2) years. There were 102 healthy children in the control group, 53 males and 49 females, with the age of (11.2±1.8) years.There was no statistically significant difference in gender and age between the two groups (both P>0.05). The serum 25 (OH) D level in children in the POTS group was (37±18)nmol/L, lower than that in the control group (61±17)nmol/L ( P<0.05).Pearson correlation analysis showed that serum 25(OH)D was positively correlated with VLF, SDNN and HF ( r=0.43, 0.65, 0.36, respectively, all P<0.05). Among the POTS children, there were 48 cases in responsive group and 17 cases in non-responsive group. Univariate analysis revealed that responsive group exhibited significantly lower values in age, SDNN, SDANN, and VLF compared to the non-responsive group (all P<0.05). Multivariate Logistic regression analysis showed that age ( OR=1.47, 95% CI 1.08-2.01), SDNN ( OR=1.01, 95% CI 1.00-1.22) and VLF ( OR=1.01, 95% CI 1.00-1.15) were independent influencing factors in the effect of vitamin D in POTS children (all P<0.05). ROC curve analysis showed that the area under the curve of VLF predicting response to vitamin D treatment for POTS was 0.72 (95% CI 0.58-0.85, P=0.008). When the cutoff value was 1 272 ms 2, the sensitivity and specificity were both 70%. Conclusion:VLF may help predict the efficacy of vitamin D treatment in children with POTS.

Objective:To explore the effect of NOD-like receptor thermal protein 3 ( NLRP3) knockout in γ-aminobutyric acid (GABA)-ergic neurons in the hippocampal CA1 area on improving cognitive dysfunction in mice after traumatic brain injury (TBI). Methods:Forty-eight healthy male NLRP3 flox/flox mice weighing 25-28 g were randomly divided into 4 groups ( n=12): sham-operated+control virus group (SV group), sham-operated+ NLRP3 specific knockout group (SG group), TBI+control virus group (TV group), TBI+ NLRP3 specific knockout group (TG group). TBI in the TV and TG groups was established by free-fall method, while surgical procedures such as scalp incision and cranial window opening without impact were given to the SV and SG groups. Adenovirus was injected into the hippocampal CA1 area of SG and TG groups 21 d before TBI to induce NLRP3 specific knockout in GABA-ergic neurons in the hippocampal CA1 area; empty virus was injected into the CA1 area of SV and TV groups. Cognitive function was evaluated using novel object recognition test 30 and 31 d after TBI, and learning and memory functions were assessed using Morris water maze test 32-36 d after TBI. Field potentials in the hippocampal CA1 area were recorded during novel object recognition 31 d after TBI. After behavioral tests, these mice were sacrificed. Immunofluorescent staining was used to detect the fluorescent intensity of microtubule-associated protein2 (MAP2), glutamic acid decarboxylase 67 (GAD67), and postsynaptic density protein 95 (PSD95) in the hippocampal CA1 area, as well as percentage of pyroptosis-associated inflammatory factor interleukin-18 (IL-18)/GAD67 double-positive neurons in total GAD67 positive neurons. Results:Compared with the SV and SG groups, the TV and TG groups had decreased novel object recognition index, decreased number of platform crossings during the experimental period, increased escape latency on day 3 and day 4 of the training period in Morris water maze test, decreased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, decreased fluorescent intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, increased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Compared with the TV group, the TG group had increased novel object recognition index, increased number of platform crossings in Morris water maze test, decreased escape latency during the training period, increased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, increased fluorescence intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, decreased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Conclusion:Specific inhibition of NLRP3 expression in GABA-ergic neurons in the hippocampal CA1 area can improve cognitive dysfunction in mice after TBI, whose mechanism may be related to inhibited GABA-ergic neuronal pyroptosis in the hippocampal CA1 area.

Objective To investigate the association of proton pump inhibitors(PPIs)use with short-term and long-term mortality risk in patients with severe ischemic stroke.Methods This retrospective study based on the U.S.Medical Information Mark for Intensive Care Ⅲ(MIMIC-Ⅲ)database,ICU patients aged ≥18 years with the first ICU admission and a diagnosis of ischemic stroke were finally included in the study.All enrolled subjects were divided into PPIs group and non-PPIs group according to whether they had used PPIs(pantoprazole,lansoprazole and omeprazole)during hospitalization.Kaplan-Meier survival analyses and Cox regression models were used to analyze the association between the use of PPIs and the risk of ICU death,30 d risk of death,90 d risk of death in patients with severe ischemic stroke.Results A total of 1 015 patients were included,402 cases in the PPIs group and 613 in the non-PPIs group.The ICU-mortality,30 d and 90 d mortality were 15.37％,13.60％and 20.10％,respectively.Kaplan-Meier survival analyses illustrated that the PPIs group survived better than non-PPIs group in ICU mortality analysis(P=0.002).In Cox regression analysis,after adjustment for potential confounders,the hazard ratio(HR)for ICU mortality in the PPIs group relative to the non-PPIs group was 0.671 9(95％CI 0.478 8 to 0.942 8,P=0.021),but there was no significant difference between 30 d and 90 d mortality(P＞0.05).Conclusion In patients with severe ischemic stroke,the use of PPIs may be effective in reducing the risk of ICU death,but does not improve 30 d and 90 d risk of death in patients.

Objective:To evaluate the role of astrocytic NOD-like receptor protein 3 (NLRP3) in the lateral hypothalamus (LHA) in anxiety-like behaviors after hemorrhagic shock and resuscitation in mice.Methods:Forty-eight clean-grade male C57BL/6 mice, aged 10 weeks, weighing 25-30 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group C), hemorrhagic shock and resuscitation group (group H), hemorrhagic shock and resuscitation + adeno-associated virus group (group HI), and hemorrhagic shock and resuscitation + control virus group (group HIV). The model of hemorrhagic shock and resuscitation was developed by bleeding and re-transfusion through the femoral vein in H, HI and HIV groups. At 21 days before developing the model, AAV-GfaABC1D-EGFP-Cre was injected into bilateral LHA in group HI, and AAV-GfaABC1D-EGFP was administered as a control in group HIV. Anxiety-like behaviors were evaluated by EPM-maze and bead-burying tests at 14 days after resuscitation. Mice were immediately sacrificed at the end of behavioral tests, and LHA-containing brain tissues were obtained for determination of co-localization of NLRP3 with glial fibrillary acidic protein (GFAP), the fluorescence intensity of Wisteria floribunda agglutinin was measured using immunofluorescent staining to reflect the expression of extracellular matrix in the LHA, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells in total cells was calculated. Results:Compared with group C, the number of buried beads and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was down-regulated, and the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was increased in H, HI and HIV groups, and the co-localization coefficient of NLRP3 and GFAP was significantly decreased in group HI ( P<0.01). Compared with group H, the number of buried particles and percentage of time of staying at the open arm were significantly decreased, the expression of extracellular matrix in the LHA was up-regulated, the co-location coefficient of NLRP3 and GFAP was decreased, the percentage of cleaved caspase-1/GFAP and IL-18/GFAP positive cells was decreased ( P<0.01), and no significant change was found in the parameters mentioned above in group HIV ( P>0.05). Conclusions:Anxiety-like behaviors after hemorrhagic shock and resuscitation is associated with astrocytic NLRP3-induced pyroptosis in the LHA and reduction of extracellular matrix in mice.

Objective:To analyze the prevalence and genotypes of human pegivirus-1 (HPgV-1) among HIV-1-infected men who have sex with men (MSM) in Zhuhai, aiming to elucidate the impact of HPgV-1 on the progression of AIDS.Methods:This study collected 934 serum specimens positive for antibodies against HIV-1 for viral RNA extraction from MSM in Zhuhai from 2012 to 2020. HPgV-1 5′UTR was amplified by nested PCR and then E gene was amplified by nested PCR and sequenced in the 5′UTR-positive specimens. A phylogenetic tree was constructed to analyze genotype distribution. The influence of HPgV-1 infection on the progression of AIDS was evaluated through analyzing HIV-1 viral load and CD4 + cell counts in patients in the early stage of AIDS before antiviral treatment. Results:The positive rate of HPgV-1 in MSM with HIV-1 infection in Zhuhai was 31.05%. A total of 273 valid sequences were obtained after amplification. The main genotype of HPgV-1 was G3 (252, 92.31%), which was highly homologous to the epidemic strains in China and Japan in recent years, followed G2 (21, 7.69%), which was highly homologous to the epidemic strains in France and America. HPgV-1 strains of G1, G4, G5, G6 and G7 genotypes were not detected. There was no significant difference in HIV-1 virus load or CD4 + cell counts between patients with HIV-1 infection alone and those with HIV-1 and HPgV-1 (G3 or G2 genotype) co-infection. Conclusions:According to the data of this study, HPgV-1 infection could not delay the progression of AIDS in MSM in the early stage of AIDS before antiviral therapy. The widespread HPgV-1 of G3 genotype in China did not have a significant impact on the progression of AIDS. Therefore, a systematic in-depth research on various genotypes of HPgV-1 and further study on the pathogenic mechanism of HPgV-1, especially in patients with HPgV-1 and HIV co-infection, were needed to understanding the interaction mechanism between different genotypes of HPgV-1 and HIV-1.

Objective:To investigate the curative effect of the Pavlik harness and polyester material casting bandage for the early treatment of congenital knee dislocation (CDK).Methods:The clinical data of 12 patients (13 knees) with CDK treated in Provincial Hospital Affiliated to Shandong First Medical University from December 2011 and January 2019 were studied retrospectively.There were 3 males and 9 females.The median age at the initial visit was 3 days ( 1-29 days). Eleven patients had unilateral CDK, and 1 patient had bilateral CDK.There were 5 cases(5 knees) with dislocation and 7 cases(8 knees) with subluxation.Associated musculoskeletal anomalies were observed in 7 patients, which included developmental dysplasia of the hip (DDH) in 6 patients, bilateral DDH and right clubfoot in 1 patient.The Pavlik harness was applied to keep the knee in position for 1.5-2.0 months if passive flexion achieved 90°or more.If passive flexion was below 90°, the polyester material casting bandage was employed to increase knee flexion, and the bandage was changed once a week.After passive flexion reached 90°, the patient was treated with the Pavlik harness for 1.5-2.0 months.Cases with DDH who worn the Pavlik harness were evaluated by ultrasonography in the follow-up period, and cases with clubfoot were treated with Ponseti polyester material casting bandages.Results:Three children (4 knees) were directly fixed with Pavlik harness for 1.5-2.0 months, 9 children (9 knees) were fixed with Pavlik harness for 1.5-2.0 months after 1-8 times (average 2.9 times) of plaster fixation. No plaster and Pavlik harness complications were found during the treatment.Twelve patients were followed up for 29.8 months on average (12.0 to 65.0 months). Ten patients (11 knees) were rated excellent, and 2 patients (2 knees) were rated good.Conclusions:The Pavlik harness and polyester material casting bandage for early treatment of CDK is simple, effective and safe.Associated musculoskeletal anomalies can also be corrected in the treatment process.The knee joint dislocation should be treated at first.

Objective To investigate the preventive and adverse effects of postnatal inhalation of Budesonide in early stage on bronchopulmonary dysplasia (BPD) in very low birth weight infants.Methods A total of 105 cases of high risk premature infants with BPD,who were born in the Neonatal Intensive Care Unit (NICU) from Shenzhen Maternity and Child Healthcare Hospital from July 15,2015 to December 25,2016,and their gestational age ≥ 27 weeks and ＜ 32 weeks or birth weight ≥ 1 000 g and ＜ 1 500 g were collected for a prospective randomized controlled trial,and were randomly divided into 3 groups:early inhalation group(34 cases),late inhalation group(34 cases) and non-inhalation group(37 cases).The oxygen time,and the incidence of BPD,periventricular-intraventricular hemorrhage (IVH),retinopathy of prematurity (ROP),necrotizing enterocolitis of the newborns (NEC),patent ductus arteriosus in preterm infants (PDA),sepsis and hyperglycemia of infants in 3 groups were compared.Results The average oxygen time in early inhalation group was 9 days,while in late inhalation group and the non-inhalation group was 15 days and 18 days,respectively.The average oxygen time in early inhalation group was significantly lower than that in the late inhalation group and the non-inhalation group,with the difference being statistically significant (H =6.09,P ＜ 0.05).The noninvasive ventilation time in early inhalation group was 3 days,while both the late inhalation group and non-inhalation group were 6 days.The noninvasive ventilation time in early inhalation group was significantly lower than that in the late inhalation group and non-inhalation group,with the difference being statistically significant (H =6.17,P ＜0.05).The incidence of BPD in the early inhalation group,late inhalation group and non-inhalation group were 14.7％ (5/34 cases),20.6％ (7/34 cases) and 37.8％ (14/37 cases),respectively.The incidence of BPD in non-inhalation group was significantly higher than that in the early inhalation group and late inhalation group,with the difference being statistically significant (x2 =12.017,P ＜ 0.05).There were no significant differences in IVH,ROP,NEC,PDA,sepsis and hyperglycemia among the 3 groups (all P ＞ 0.05).Conclusions Postnatal inhalation of Budesonide in early stage in high risk very low birth weight infants can reduce the incidence of BPD and the oxygen time,and the adverse reactions are not obvious.