Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

A 75-year-old male patient with non-small cell lung cancer in the lower lobe of the left lung underwent left lobectomy, followed by 4 cycles of adjuvant chemotherapy with paclitaxel and platinum, 4 cycles of chemotherapy with docetaxel and cisplatin, and immunotherapy with sintilimab, 13 cycles of immunotherapy with sintilimab monotherapy, and 5 cycles of endostatin and sintilimab, spanning a total of 5 years. Due to disease progression, he received the monotherapy with anlotinib, 42 days later, the patient developed intestinal perforation, and laparoscopic ileal perforation repair surgery was performed. During the operation, a perforation with a diameter of about 1 cm was observed in the ileum, indicating the outflow of digestive fluid and a large amount of purulent fluid accumulation in the pelvic cavity; no signs of gastrointestinal tumor metastasis were observed. After surgery, the patient developed abdominal infection and peritonitis, and was given symptomatic and supportive treatments such as anti-infection for 7 days. However, the patient developed severe complications such as lung infection, heart failure, sepsis, and respiratory failure, and died.

A 75-year-old male patient with non-small cell lung cancer in the lower lobe of the left lung underwent left lobectomy, followed by 4 cycles of adjuvant chemotherapy with paclitaxel and platinum, 4 cycles of chemotherapy with docetaxel and cisplatin, and immunotherapy with sintilimab, 13 cycles of immunotherapy with sintilimab monotherapy, and 5 cycles of endostatin and sintilimab, spanning a total of 5 years. Due to disease progression, he received the monotherapy with anlotinib, 42 days later, the patient developed intestinal perforation, and laparoscopic ileal perforation repair surgery was performed. During the operation, a perforation with a diameter of about 1 cm was observed in the ileum, indicating the outflow of digestive fluid and a large amount of purulent fluid accumulation in the pelvic cavity; no signs of gastrointestinal tumor metastasis were observed. After surgery, the patient developed abdominal infection and peritonitis, and was given symptomatic and supportive treatments such as anti-infection for 7 days. However, the patient developed severe complications such as lung infection, heart failure, sepsis, and respiratory failure, and died.

Objective:To investigate the effect of Stigma Maydis Palysaccharide(SMPS)on ATP synthesis in kidney mitochondria of D-galactose-induced aging mice, and to clarify its possible mechanism.Methods:The aging mouse model was established by subcutaneous injection of D-galactose solution in the back of the neck.The 48 SPF male mice were randomly divided into normal control group(control group), D-galactose model group(D-Gal group), SMPS low-dose group and SMPS high-dose group(n=12 for each). The control group was subcutaneously injected with 150 mg/kg normal saline on the back of the neck every day, while the other three groups were subcutaneously injected with 150 mg/kg of D-gal solution on the back of the neck every day.SMPS-L and-H dose groups were given 30 mg/kg and 60 mg/kg of SMPS solution by gavage at the same day of D-Gal injection.The control group and D-GAL group were given the same volume of normal saline daily by gavage for 42 days.Blood samples were collected from the eyeball under general anesthesia after 42 days of intervention for the detection of serum levels of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px)and MDA.After harvesting the kidney tissue, various tests were used to detect ATP content, the mRNA expression levels and protein expression levels in kidney.Luciferase assay was used to detect ATP content in renal tissue.Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of succinate dehydrogenase(SDH)of complex Ⅱ, cytochrome C reductase(Cycs)of complex Ⅲ, complex Ⅳ(COXⅣ)and ATP5b in ATP synthase in mitochondrial oxidative respiratory chain.Western blot was used to detect the expression levels of mitochondrial fusion protein 2(MFN2), dynamin-related protein1(DRP1)and mitochondrial autophagy related protein P62 in renal tissues of each group.Results:Compared with control group, the activities of serum of SOD(116.53±10.01)U/mg and GSH-Px(127.58±8.74)μmol/L were significantly decreased in D-GAL group(both P< 0.01), and serum MDA content(15.42±0.91)μmol/L increased significantly in D-GAL group( P<0.01). Compared with D-GAL group, the activities of SOD(152.80±9.29)U/mg and GSH-Px(274.07±10.73)μmol/L were significantly increased in SMPS intervention group( P< 0.01), while the MDA content(8.10±0.66)μmol/L decreased significantly in SMPS intervention group( P< 0.01). Compared with control group, the content of ATP(178±4)10 -4 μmol in D-gal group was significantly decreased( P<0.01), the mRNA expression levels of SDH, Cycs and COXⅣ were not significantly changed in D-gal group, and the mRNA expression level of ATP5b(0.67±0.01)was down-regulated in D-gal group( P<0.01), the expression of MFN2 protein(0.29±0.02)was significantly decreased in D-gal group( P<0.01), and the expression of DRP1 and P62 protein(0.31±0.02 and 0.21±0.01)was significantly increased in D-gal group(both P<0.01). Compared with the D-gal group, the ATP content(193±1)10 -4 μmol in the kidney tissue of the mice was significantly increased in SMPS intervention group( P< 0.01), and the ATP5b mRNA expression and MFN2 protein expression(0.87±0.05 and 0.71±0.08)were significantly increased in SMPS intervention group(both P< 0.01), DRP1 and P62 protein expressions(0.20±0.01 and 0.10±0.01)were significantly down-regulated in in SMPS intervention group(both P< 0.01). Conclusions:SMPS can improve the mitochondrial dynamic homeostasis disorder in aging mice by increasing the activity of antioxidant enzymes, up-regulating the expression of ATP5b mRNA and MFN2 protein, down-regulating the expression of DRP1 and P62 protein, and promoting the generation of mitochondrial ATP in D-gal-induced aging mice kidney tissue.

Helicobacter pylori (Hp) resides in the epithelium of gastric mucosa and can cause chronic inflammation. Under the conventional white light endoscopy, normal gastric mucosa, current Hp-infected gastric mucosa and past Hp-infected gastric mucosa have different endoscopic features. High-resolution white light endoscopy and various new image-enhanced endoscopy are helpful to accurately diagnose the infection of Hp, and thereby improving the detection rate of early gastric cancer by identifying the high-risk endoscopic manifestations related to gastric cancer. This article reviewed the endoscopic diagnosis of Hp infection and its clinical value.

Objective: Children with autism spectrum disorder(ASD) who participated in the " Play-based Communication and Behavior Intervention(PCBI)" were studied in order to evaluate the impact of caregiver factors on the efficacy of the PCBI very early intervention model systematically. Methods: Seventy-six children with ASD aged 8-30 months were recruited in this study at Brain Hospital Affiliated to Nanjing Medical University form December 2017 to June 2018, and they were trained for PCBI very early intervention.The Autism Treatment Evaluation Checklist (ATEC) and the Gesell Developmental Schedules were used to evaluate the efficacy of the PCBI very early intervention.Parenting Stress Index Short Form, General Self-Efficacy Scale and the self-developed Caregiver Training Course Evaluation Scale and the Homework Completion Level Scale were used to assess the level of parental stress, self-efficacy, curriculum satisfaction and acceptance.A pair of sample t-test was used to analyze the ASD symptoms, developmental level and caregiver factors of ASD children before and after intervention.Multivariate regression analysis was used to analyze the caregiver factors and the effects of PCBI very early intervention. Results: (1) At the end of 12 weeks of PCBI very early intervention, the score of the ATEC (17.36±15.98) was lower than that before intervention (78.43±22.84), and the difference was statistically significant(P<0.05); the adaptive developmental quotient(5.90±15.45), the fine action developmental quotient(5.13±19.89), the language developmental quotient (12.43±20.60) and the personal social developmental quotient(4.79±16.02) scores of the Gesell Developmental Schedules were higher than those before intervention (73.08±20.54, 77.33±23.63, 52.24±19.12, 71.79±20.81), and the differences were statistically significant (all P<0.05). (2) At the end of PCBI very early intervention, there was a significant decrease in the scores of Parenting Stress Index Short Form, and the differences were statistically significant (4.84±14.69 vs.94.05±29.67, P<0.05). (3) Acceptance of the intervention model, and mastery of the skills in the caregiver factors had a positive predictive effect on the effect of PCBI very early intervention (t=3.068, 10.468, all P<0.05). Conclusions PCBI very early intervention can effectively reduce the parenting pressure of the caregivers, and in the PCBI very early intervention, the better the mastery of skills and the completion of homework, the better the efficacy of intervention.

Objective To explore the characteristics of social behaviors in toddlers with high-risk of autism spectrum disorder (HR-ASD),and the correlation to the severity of ASD symptoms.Method 32 high-risk ASD toddlers,aged from 12 months to 24 months,and 34 typical developmental (TD) infants/toddlers,aged from 6 months to 24 months,were recruited.Behavioral video of the Still-Face Paradigm (SFP) was used to measure the frequencies and duration of social behaviors,including non-social smiling,protest,eye contact,social smiling and social positive engagement,during the mother-infant interaction and the still face.Autism Diagnostic Observation Scale was used to evaluate severity of clinical symptoms in high-risk ASD toddler.Gesell Development Scale was used to assess the developmental level of the two groups.Independent-sample t test was used to compare the differences of social behaviors between two groups.Pearson correlation analysis was used to analyze the correlation between the social behaviors of high-risk ASD and the age,the developmental level and the severity of ASD symptoms.Binary logistic regression was used to analyze the power of discrimination of social behaviors between HR-ASD and TD.Results During mother-infant interaction,compared with TD group,HR-ASD had significantly shorter duration ((19.8±23.4) s vs.(61.5±34.0) s,t=-5.75,P＜0.01;(6.5±9.3) s vs.(29.0±25.2) s,t=-4.73,P＜0.01;(3.0±8.8) s vs.(13.2±23.1) s,t=-2.34,P=0.02) and lower frequencies (t=-6.62、-3.03、-2.74,all P＜0.01) of eye contact,social smiling and social positive engagement.During the still face,compared with TD group,HR-ASD had significantly shorter duration (t=-3.32,P＜0.01;t=-2.59,P=0.01) and lower frequencies (t=-6.13、-3.48,all P＜0.01) of eye contact and social positive engagement.According to Pearson correlation analysis,during the still face,in HR-ASD group,the duration of eye contact and social positive engagement had significantly negative influence on the score of the language and communication in ADOS (r=-0.49、-0.52,all P＜0.01);The duration and frequencies of eye contact and social positive engagement had significantly negative influence on the score of mutually social interaction in ADOS (r=-0.57,P＜0.01;r=-0.49,P＜0.01;r=-0.74,P＜0.01;r=-0.44,P=0.01);The duration of eye contact and social positive engagement had significantly negative influence on the score of the playing in ADOS (r=-0.43,P=0.01;r=-0.45,P=0.01).According to binary logistic regression,the duration of social smiling during the still face and the frequencies of eye contact during mother-infant interaction could distinguish between HR-ASD and TD (OR=1.36,95％CI:1.08-1.71,P=0.01;OR=5.99,95％CI:1.15-31.14,P=0.03).Conclusion HR-ASD hasthe lower levels of eye contact,social smiling and social positive engagement,which may suggestthe severity of ASD core symptoms.The duration of social smiling during the still face and the frequencies of eye contact during mother-infant interaction may possiblyfigure out theHR-ASD and could be further developed as a predictablebehavioral maker.

Objective To explore the characteristics of social behaviors in toddlers with high-risk of autism spectrum disorder (HR-ASD),and the correlation to the severity of ASD symptoms.Method 32 high-risk ASD toddlers,aged from 12 months to 24 months,and 34 typical developmental (TD) infants/toddlers,aged from 6 months to 24 months,were recruited.Behavioral video of the Still-Face Paradigm (SFP) was used to measure the frequencies and duration of social behaviors,including non-social smiling,protest,eye contact,social smiling and social positive engagement,during the mother-infant interaction and the still face.Autism Diagnostic Observation Scale was used to evaluate severity of clinical symptoms in high-risk ASD toddler.Gesell Development Scale was used to assess the developmental level of the two groups.Independent-sample t test was used to compare the differences of social behaviors between two groups.Pearson correlation analysis was used to analyze the correlation between the social behaviors of high-risk ASD and the age,the developmental level and the severity of ASD symptoms.Binary logistic regression was used to analyze the power of discrimination of social behaviors between HR-ASD and TD.Results During mother-infant interaction,compared with TD group,HR-ASD had significantly shorter duration ((19.8±23.4) s vs.(61.5±34.0) s,t=-5.75,P＜0.01;(6.5±9.3) s vs.(29.0±25.2) s,t=-4.73,P＜0.01;(3.0±8.8) s vs.(13.2±23.1) s,t=-2.34,P=0.02) and lower frequencies (t=-6.62、-3.03、-2.74,all P＜0.01) of eye contact,social smiling and social positive engagement.During the still face,compared with TD group,HR-ASD had significantly shorter duration (t=-3.32,P＜0.01;t=-2.59,P=0.01) and lower frequencies (t=-6.13、-3.48,all P＜0.01) of eye contact and social positive engagement.According to Pearson correlation analysis,during the still face,in HR-ASD group,the duration of eye contact and social positive engagement had significantly negative influence on the score of the language and communication in ADOS (r=-0.49、-0.52,all P＜0.01);The duration and frequencies of eye contact and social positive engagement had significantly negative influence on the score of mutually social interaction in ADOS (r=-0.57,P＜0.01;r=-0.49,P＜0.01;r=-0.74,P＜0.01;r=-0.44,P=0.01);The duration of eye contact and social positive engagement had significantly negative influence on the score of the playing in ADOS (r=-0.43,P=0.01;r=-0.45,P=0.01).According to binary logistic regression,the duration of social smiling during the still face and the frequencies of eye contact during mother-infant interaction could distinguish between HR-ASD and TD (OR=1.36,95％CI:1.08-1.71,P=0.01;OR=5.99,95％CI:1.15-31.14,P=0.03).Conclusion HR-ASD hasthe lower levels of eye contact,social smiling and social positive engagement,which may suggestthe severity of ASD core symptoms.The duration of social smiling during the still face and the frequencies of eye contact during mother-infant interaction may possiblyfigure out theHR-ASD and could be further developed as a predictablebehavioral maker.

OBJECTIVE: To determine the association between carotid atherosclerosis and renal function in hypertensive patients. METHODS: A total of 2 809 hypertensive patients aged (56.59±10.79) years were enrolled. Carotid intima-media thickness (cIMT) was derived via B-mode ultrasonography and chronic kidney disease (CKD) was evaluated by the estimated glomerular filtration rate (eGFR) with Cockcroft- Gault method. The patients were divided into 3 groups: a normal group, a thick group, and a plaque group according to the results of carotid ultrasonography. RESULTS: The eGFR of the normal group was (111.09±25.61) mL/(min.1.73m(2)), that of the thick group and the plaque group was (94.45±27.14) mL/(min.1.73m(2)) and (85.98±26.92) mL/ (min.1.73m(2)). Binary logistic analysis showed that age (OR=3.590), smoking status (OR=1.543), systolic blood pressure (OR=1.018), diastolic blood pressure (OR=0.977), fasting plasma glucose (OR=1.132), triglyceride (OR=0.873) and eGFR (OR=0.986) were significantly correlated with cIMT. Subgroup analyses on different genders showed that eGFR was a significant independent risk factor in men (OR=0.991) but not in women. CONCLUSION The thicker the cIMT, the lower the eGFR in hypertensive patients. With the development of cIMT, eGFR gradually decreases and contributes to the occurrence and development of early-stage atherosclerosis in hypertensive patients.
Aged ; Atherosclerosis ; Blood Pressure ; Carotid Arteries ; diagnostic imaging ; pathology ; Carotid Artery Diseases ; Carotid Intima-Media Thickness ; Female ; Glomerular Filtration Rate ; Humans ; Hypertension ; physiopathology ; Male ; Middle Aged ; Renal Insufficiency, Chronic ; Risk Factors ; Smoking

Objective To explore the clinical nursing effect of the atomization inhalation therapy on pediatric asthmatic suffocating pneumonia .Methods Ninety-three patients with pediatric asthmatic suffocating pneumonia were chosen and randomly divided into the observation group ( n=47 ) and the control group ( n=46).The control group received the routine treatment (controlling infection through antibiotics , keeping calm and reducing of asthma , relieving cough and sputum , etc.) and conventional nursing measures ( health education, diet nursing, psychological nursing, etc.), and the observation group received the atomization inhalation therapy on the basis of the control group .The nursing and therapeutic effects were compared between the two groups .Results The total effective rate of treatment was 93 .6%( 44/47 ) in the observation group , which was higher than 67.4%( 31/46 ) in the control group , and the difference was statistically significant (χ2 =10.244, P<0.05).The average time of the disappear of asthma and moist rales and the hospital stay were (6.4 ±1.5), (7.0 ±1.7), (8.1 ±2.1) d in the observation group, which were shorter than (9.1 ± 1.4), (10.2 ±2.0), (10.9 ±1.9) d in the control group, and the differences were statistically significant (t=10.541, 13.293, 10.941, respectively; P <0.01).Conclusions Application of the atomization inhalation therapy on pediatric asthmatic suffocating pneumonia based on the routine treatment and nursing can effectively improve the therapeutic effect , relieve the pain of patients and promote the rehabilitation of patients .