This study investigated the specific immune response of BALB/c mice that was induced by a circular RNA (circRNA) vaccine expressing the herpes simplex virus type II (HSV-2) glycoprotein D (gD). The aim was to evaluate the immunological potential of this vaccine and lay a foundation for developing an mRNA vaccine against HSV-2. PCR and homologous recombination were employed to integrate the gD gene obtained from the pT7AMP-gD ectodomain plasmid into pUC57 to generate the recombinant plasmid pUC57-circ-gD, which was then sequenced and characterized. In vitro transcription and cyclization were performed on the template DNA to generate pUC57-circ-gD mRNA. To validate the formation of circular RNA, we cleaved the pUC57-circ-gD mRNA with RNase R and employed RT-PCR to validate the cyclization. The pUC57-circ-gD mRNA was then transfected into 293T cells. After 72 h, the cell supernatant was collected, and Western blotting was employed to measure the protein level of gD. Subsequently, the mRNA was encapsulated in lipid nanoparticles (LNPs) by microfluidic encapsulation. BALB/c mice were administrated with the encapsulated mRNA, and blood was collected from the fundus venous plexus after 21 and 35 days, and from the enucleated eyeballs after 49 days. Enzyme-linked immunosorbent assay was employed to measure the titers of antibodies, including virus-neutralizing antibodies. After 49 days, spleens were harvested and assessed for secretion of interferon-gamma (IFN-γ) by solid-phase enzyme-linked immunospot. The results showed successful construction and sequencing of the recombinant plasmid. RNase R digestion confirmed the presence of circular RNAs. Western blotting of the 293T cells transfected with the mRNA showed clear specific bands. The quality of the vaccine was tested by size exclusion chromatography-high performance liquid chromatography, which showed that the purity of the vaccine was about 90%. The mRNA-LNP showcased the particle size of 82.76 nm and an encapsulation rate of approximately 98%. Following three-dose vaccination, all immunized mice exhibited steady weight gain with 100% survival rate throughout the 28-day observation period, indicating no significant acute toxicity associated with the vaccine formulation. The immunized mice showed dose-dependent increases in serum IgG antibody titer and IFN-γ secretion by splenocytes and they were resistant to virus attacks. These findings indicate good immunogenicity and persistence of the pUC57-circ-gD mRNA vaccine, providing a reference for further studies on circRNA vaccines.
Animals ; Mice, Inbred BALB C ; RNA, Circular ; Mice ; Humans ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Antibodies, Viral/blood* ; HEK293 Cells ; Female ; Nanoparticles ; Plasmids

ObjectiveTo understand the epidemic characteristics of influenza in Beijing from 2023 to 2024, and to provide a scientific basis for the prevention and control of influenza. MethodsData on influenza-like illness (ILI) from secondary level and above hospitals, etiology surveillance data, and influenza clusters outbreaks data from 2023‒2024 were used to analyze the epidemic trend and pathogenic characteristics of influenza. Furthermore, an influenza comprehensive index was used to categorize the epidemic intensity at the severity level. ResultsA total of 2 065 857 ILI cases were reported in 2023‒2024 epidemic season, and the percentage of ILI was 3.67%. The age group of 5‒14 years accounted for the highest proportion of ILI (30.48%). A total of 41 766 throat swabs from ILI were detected, with a positive rate of 17.28%.A (H3N2) (51.86%) and B Victoria (41.93%) were the most prevalent subtypes of influenza virus. Clustered influenza outbreaks occurred mainly in primary schools (57.78%) and middle schools (35.55%), mainly caused by the influenza A (H3N2) subtype (85.93%). According to the influenza comprehensive index (I), the period of influenza activity and above (I>0.5) lasted for a total of 37 weeks, accounting for 71.15% of the entire influenza season. ConclusionCompared with previous years, the epidemic level of influenza in Beijing was increased in 2023‒2024, and the peak time became earlier. The comprehensive index method can objectively evaluate the level of influenza epidemic and provide suggestions for the future prevention and control of influenza in Beijing.

Objective:To investigate the relationship between thyroid hormone levels during pregnancy and neonatal thyroid function.Methods:Forty pregnant women with abnormal thyroid stimulating hormone (TSH) level during pregnancy and their newborns who received treatment in Yongkang First People's Hospital, China between July 2019 and August 2020 were included in the observation group. An additional 38 healthy pregnant women and their newborns who concurrently received health examination were included in the control group. The clinical data in the two groups were retrospectively analyzed. The levels of thyroid hormones [(triiodothyronine (T 3), tetraiodothyronine (T 4), TSH, free T 3 (FT 3), free T 4 (FT 4)] in pregnant women were compared between the two groups. TSH level in newborns was compared between the two groups. Thyroid dysfunction in newborns was assessed in each group. The correlation between thyroid hormone levels during pregnancy and neonatal thyroid function was analyzed. Results:TSH level during pregnancy in the observation group was significantly higher than that in the control group [(2.89 ± 0.44) mU/L vs. (2.13 ± 0.22) mU/L, t = 9.570, P < 0.001]. T 3, T 4, FT 3 and FT 4 in the observation group were (0.45 ± 0.07) μg/L, (90.87 ± 8.93) μg/L, (1.08 ± 0.19) ng/L and (10.45 ± 1.73) ng/L, respectively, which were significantly lower than those in the control group [(2.13 ± 0.22) μg/L, (1.31 ± 0.21) μg/L, (2.16 ± 0.34) ng/L, (15.31 ± 21) ng/L, t = 24.514, 9.254, 17.432, 10.845, all P < 0.001]. TSH level in newborns in the observation group was significantly higher than that in the control group ( t = 37.041, P < 0.05). The incidence of thyroid dysfunction in the observation group was significantly higher than that in the control group ( χ2 = 4.780, P < 0.05). TSH level in pregnant women was positively correlated with that in newborns ( r = 0.819, P < 0.05). T 3, T 4, FT 3 and FT 4 levels in pregnant women were negatively correlated with TSH level in newborns ( r = -0.773, -0.802, -0.794, -0.824, all P < 0.05). Conclusion:Compared with healthy pregnant women, pregnant women with abnormal thyroid hormone levels have higher TSH levels and lower T 3, T 4, FT 3 and FT 4 levels. The newborns of pregnant women with abnormal thyroid hormone levels have higher TSH levels and a greater risk of thyroid dysfunction than the newborns of healthy pregnant women. The level of thyroid hormone during pregnancy is related to the thyroid function of newborns. This study is scientific and innovative.

Objective:To summarize experience of nursing in radiofrequency ablation (RFA) for adrenocortical adenoma with Cushing′s syndrome combined steroid-induced diabetes mellitus.Methods:The key of preoperative nursing was precise glucose control by using insulin pump. Focus on patients psychological intervention. It was important for closely monitor the change of blood pressure, cortisol levels and biochemical index to prevent acute complications such as adrenal crisis during RFA. Pay attention to postoperative nutrition management for reduce blood glucose fluctuation and adverse reactions of hydrocortisone at postoperative.Results:No severe complications occurred during this procedure. After twenty-day nursing and treatment, the patient′s blood glucose control was stable, and the symptoms related to Cushing′s syndrome gradually disappeared.Conclusion:The ultrasound guided radiofrequency ablation in adrenocortical adenoma with Cushing′s syndrome combined with steroid-induced diabetes mellitus was a feasible, safe, and minimally invasive treatment. The key to effectively reduce postoperative complications and ensure the success of the operation. Are adequate preoperative preparation and strict monitor.

Objective:Assess the relationship between elevated antiphospholipid antibodies and thrombosis in hospitalized patients.Methods:Case control study. A total of 385 patients (149 males and 236 females, aged from 1 to 105 years, with a median age of 37 years) who were hospitalized in Peking University First Hospital from January 2015 to December 2019 and tested positive for any one of the anti-phospholipid antibodies were included in the study. All subjects were divided into thrombotic group and non-thrombotic group according to whether thrombus was detected by imaging examination during hospitalization. In thrombosis group, there were 66 males and 36 females, aged from 3 to 105 years, with a median age of 58 years. In non-thrombosis group, there were 83 males and 200 females, aged from 1 to 94 years, with a median age of 31 years. Clinical data and laboratory data of patients were recorded. ACL-IgM/IgG and anti-β2GPI-IgM/IgG were detected by ELISA and LA was detected by dRVVT and SCT on automatic coagulation analyzer. The rates of age, gender, smoking, obesity, hypertension, hyperlipidemia, diabetes and the median level of antiphospholipid antibodies were compared between two groups. Logistic multivariate regression analysis was used to determine the risk factors for thrombotic events. The mid-to-high titer value of aCL was established by the χ 2-trend test and verified by logistic regression. Results:The median age (58 years) and the rates of male (64.7%), smoking (16.7%), hypertension (63.7%) and diabetes (28.4%) in thrombus group were significantly higher than those in non-thrombus group ( Z=7.685, χ2=38.077, 16.312, 37.769, 24.749 respectively; P<0.01). The positive rate of anti-β2GPI-IgG and dRVVT in thrombosis group (11.8% and 78.4%) was significantly higher than that in non-thrombosis group (5.3% and 60.1%), as well as the median level of dRVVT (1.29 RU/ml vs 1.23 RU/ml) (χ2=3.864 and 10.309, Z=3.539; P<0.05). The median level of aCL-IgM was higher in non-thrombosis group (2.3 MPL vs 2.0 MPL). The positive rate of aCL-IgG was slightly higher in thrombosis group (18.6% vs 10.6%). Logistic regression analysis showed that men, hypertension, diabetes, advanced age, elevated dRVVT, and elevated anti-β2GPI-IgG are risk factors for thrombosis. Taking 36 GPL as the medium-to-high titer value of aCL-IgG, the risk of thrombosis increased by 2.45 times. Conclusions:In the anti-phospholipid antibody profile, LA detected by dRVVT method, anti-β2GPI-IgG and aCL-IgG may be valuable laboratory indicators for inpatient thrombotic events. The mid-to-high titer value of aCL-IgG is set at 36 GPL to distinguish the risk of thrombosis.

Objective:Assess the relationship between elevated antiphospholipid antibodies and thrombosis in hospitalized patients.Methods:Case control study. A total of 385 patients (149 males and 236 females, aged from 1 to 105 years, with a median age of 37 years) who were hospitalized in Peking University First Hospital from January 2015 to December 2019 and tested positive for any one of the anti-phospholipid antibodies were included in the study. All subjects were divided into thrombotic group and non-thrombotic group according to whether thrombus was detected by imaging examination during hospitalization. In thrombosis group, there were 66 males and 36 females, aged from 3 to 105 years, with a median age of 58 years. In non-thrombosis group, there were 83 males and 200 females, aged from 1 to 94 years, with a median age of 31 years. Clinical data and laboratory data of patients were recorded. ACL-IgM/IgG and anti-β2GPI-IgM/IgG were detected by ELISA and LA was detected by dRVVT and SCT on automatic coagulation analyzer. The rates of age, gender, smoking, obesity, hypertension, hyperlipidemia, diabetes and the median level of antiphospholipid antibodies were compared between two groups. Logistic multivariate regression analysis was used to determine the risk factors for thrombotic events. The mid-to-high titer value of aCL was established by the χ 2-trend test and verified by logistic regression. Results:The median age (58 years) and the rates of male (64.7%), smoking (16.7%), hypertension (63.7%) and diabetes (28.4%) in thrombus group were significantly higher than those in non-thrombus group ( Z=7.685, χ2=38.077, 16.312, 37.769, 24.749 respectively; P<0.01). The positive rate of anti-β2GPI-IgG and dRVVT in thrombosis group (11.8% and 78.4%) was significantly higher than that in non-thrombosis group (5.3% and 60.1%), as well as the median level of dRVVT (1.29 RU/ml vs 1.23 RU/ml) (χ2=3.864 and 10.309, Z=3.539; P<0.05). The median level of aCL-IgM was higher in non-thrombosis group (2.3 MPL vs 2.0 MPL). The positive rate of aCL-IgG was slightly higher in thrombosis group (18.6% vs 10.6%). Logistic regression analysis showed that men, hypertension, diabetes, advanced age, elevated dRVVT, and elevated anti-β2GPI-IgG are risk factors for thrombosis. Taking 36 GPL as the medium-to-high titer value of aCL-IgG, the risk of thrombosis increased by 2.45 times. Conclusions:In the anti-phospholipid antibody profile, LA detected by dRVVT method, anti-β2GPI-IgG and aCL-IgG may be valuable laboratory indicators for inpatient thrombotic events. The mid-to-high titer value of aCL-IgG is set at 36 GPL to distinguish the risk of thrombosis.

Objective: To investigate the role of adenovirus type 36 (Ad36) in the browning of 3T3-L1 cells. Methods: BODIPY staining was performed on the 0, 2nd, 4th, 6th and 8th days of the cocktail induction (control) group and the cocktail plus Ad36 induction (experimental) group to observe the adipogenesis of 3T3-L1 cells.The mRNA and protein expressions of uncoupling protein-1(Ucp1), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit (Atp5o), cytochrome c oxidase subunit 5B(Cox5b), and perilipin were detected by real-time PCR and Western-blot. Results: The results of BODIPY staining showed that the lipid droplets in the control group gradually became larger with the differentiation of the cells, while the lipid droplets in the experimental group firstly became larger, and then appeared smaller after Ad36 was added on the fourth day. Compared with the control group, the mRNA and protein expression levels of Ucp1, Atp5o, and Cox5b in the experimental group were significantly increased while the expression level of perilipin was significantly decreased (all P<0.05). Conclusion During the differentiation of 3T3-L1 cells, Ad36 promotes its browning via increasing the expressions of Ucp1, Atp5o, and Cox5b.

Objective To investigate the role of LncRNA ROR in Ad36-induced browning of human adipose-derived stem cells(hADSC). Methods After hADSC was induced by cocktail and Ad36 for 2,4,6,and 8 days,Oil red O staining was performed for observing the adipogenic status. The mRNA expressions of LncRNA ROR, uncoupling protein 1(UCP1),and PRDM16 were detected by real-time PCR and the protein expressions of UCP1 and PRDM16 were detected by Western-blot. After LncRNA ROR was knocked down by siRNA, UCP1 and PRDM16 mRNA and protein expression levels in the process of Ad36-induced adipocyte differentiation were detected by real-time PCR and Western-blot. Results Oil red O staining showed that fat droplets in the cocktail-induced group were larger than those in the Ad36-induced group. Compared with the cocktail group,the mRNA expressions of LncRNA ROR, UCP1 and PRDM16, and the protein expressions of UCP1 and PRDM16 in Ad36 group were significantly increased(P<0.05). The mRNA and protein expressions of UCP1 and PRDM16 in LncRNA ROR knockdown group were significantly lower than those in control group(P<0.05). Conclusion In the process of Ad36-induced hADSC differentiation,the up-regulation of LncRNA ROR may stimulate UCP1 and PRDM16 expression,and thus promote the browning of hADSC.

Described in the paper is the reform made at public hospitals in Zhejiang province,with analysis of its problems and causes. Based on such,the authors proposed the following actions:to accelerate the reform of public hospitals centering on health promotion, and encourage such hospitals to shoulder the health care responsibility on behalf of the government; to proceed with hierarchical medical system and contract-based service focusing on capacity building of general practitioners; to attract private capital into building the healthcare system; and to motivate non-governmental organizations to cater to diversified healthcare needs of the people.

Objective To analyze of the effect of artificial lengthening femoral head replacement in elderly patients with stage Ⅰ of unstable femoral intertrochanteric fracture.Methods 203 patients with stage Ⅰ of unstable femoral intertrochanteric fracture were selected as the research object,and they were taken artificial lengthening femoral head replacement,among which 65caese were male,female in 138 cases.The Harris scoring,SF-36,VAS pain scores on admission,2 weeks after operation,postoperative follow-up limb were counted,and the pain of the affected limb and the hip scores were compared amond 3 time periods.Results All 203 cases of senile patients with follow-up,average operation time was 83.64 minutes,the intraoperative blood loss was 355.41mL.The curative effect was evaluated according to the Harris score,SF-36 and VAS pain scoring criteria,and the Harris scores of the affected limbs at admission,at 2 weeks after the operation and after the follow-up were (28.26 ± 5.50) points,(68.26 ±5.50) points,(93.13 ± 5.31) points,respectively,the differences were statistically significant (t =-71.27,-1 397.55,-46.07,all P ＜ 0.01);The VAS pain scores were (8.19 ± 0.48) points,(3.53 ± 0.71) points,(0.23 ± 0.42) points,respectively,the differences were statistically significant (t =88.06,324.17,60.84,all P ＜ 0.01).The sf-36 scores:physiological [(8.35 ± 1.24) points,(15.23 ± 2.17) points,(19.21 ± 2.12) points],social/family [(7.01 ±1.13) points,(14.12 ± 2.12) points,(19.85 ± 2.24) points],emotional [(4.83 ± 1.01) points,(10.12 ±1.22)points,(14.87 ± 1.32) points],function [(6.35 ± 1.21) points,(13.67 ± 1.87) points,(16.81 ±2.12) points],additional focus [(8.85 ± 1.45) points,(16.38 ± 2.12) points,(20.21 ± 2.42) points],total quality of life [(47.35 ± 4.76) points,(74.69 ± 5.87) points,(89.21 ± 6.12) points],the differences were statistically significant(-39.77,-62.92,-20.21,-44.87,-71.89,-26.79,-45.04,-89.01,-38.25,-45.79,-63.41,-15.29,-45.20,-60.39,-17.54,-52.12,-76.49,-22.58,all P＜0.O1).Conclusion Artificial lengthening femoral head replacement in elderly patients with stage Ⅰ of unstable femoral intertrochanteric fracture has good clinical effect,intraoperative high safety,less postoperative complications,postoperative limb functional recovery is good,and it is worthy of clinical promotion and application.