[Immunogenicity, safety and immune persistence of the sequential booster with the recombinant protein-based COVID-19 vaccine (CHO cell) in healthy people aged 18-84 years].

Objective: To evaluate the immunogenicity, safety, and immune persistence of the sequential booster with the recombinant protein-based COVID-19 vaccine (CHO cell) in healthy people aged 18-84 years. Methods: An open-label, multi-center trial was conducted in October 2021. The eligible healthy individuals, aged 18-84 years who had completed primary immunization with the inactivated COVID-19 vaccine 3 to 9 months before, were recruited from Shangyu district of Shaoxing and Kaihua county of Quzhou, Zhejiang province. All participants were divided into three groups based on the differences in prime-boost intervals: Group A (3-4 months), Group B (5-6 months) and Group C (7-9 months), with 320 persons per group. All participants received the recombinant COVID-19 vaccine (CHO cell). Blood samples were collected before the vaccination and after receiving the booster at 14 days, 30 days, and 180 days for analysis of GMTs, antibody positivity rates, and seroconversion rates. All adverse events were collected within one month and serious adverse events were collected within six months. The incidences of adverse reactions were analyzed after the booster. Results: The age of 960 participants was (52.3±11.5) years old, and 47.4% were males (455). The GMTs of Groups B and C were 65.26 (54.51-78.12) and 60.97 (50.61-73.45) at 14 days after the booster, both higher than Group A's 44.79 (36.94-54.30) (P value<0.05). The GMTs of Groups B and C were 23.95 (20.18-28.42) and 27.98 (23.45-33.39) at 30 days after the booster, both higher than Group A's 15.71 (13.24-18.63) (P value <0.05). At 14 days after the booster, the antibody positivity rates in Groups A, B, and C were 91.69% (276/301), 94.38% (302/320), and 93.95% (295/314), respectively. The seroconversion rates in the three groups were 90.37% (272/301), 93.75% (300/320), and 93.31% (293/314), respectively. There was no significant difference among these rates in the three groups (all P values >0.05). At 30 days after the booster, antibody positivity rates in Groups A, B, and C were 79.60% (238/299), 87.74% (279/318), and 90.48% (285/315), respectively. The seroconversion rates in the three groups were 76.92% (230/299), 85.85% (273/318), and 88.25% (278/315), respectively. There was a significant difference among these rates in the three groups (all P values <0.001). During the sequential booster immunization, the incidence of adverse events in 960 participants was 15.31% (147/960), with rates of about 14.38% (46/320), 17.50% (56/320), and 14.06% (45/320) in Groups A, B, and C, respectively. The incidence of adverse reactions was 8.02% (77/960), with rates of about 7.50% (24/320), 6.88% (22/320), and 9.69% (31/320) in Groups A, B, and C, respectively. No serious adverse events related to the booster were reported. Conclusion: Healthy individuals aged 18-84 years, who had completed primary immunization with the inactivated COVID-19 vaccine 3 to 9 months before, have good immunogenicity and safety profiles following the sequential booster with the recombinant COVID-19 vaccine (CHO cell).

[The expression and function of PD-L1 in CD133(+) human liver cancer stem-like cells].

Objective: To investigate the expression of programmed death protein-ligand 1 (PD-L1) in liver cancer stem-like cells (LCSLC) and its effect on the characteristics of tumor stem cells and tumor biological function, to explore the upstream signaling pathway regulating PD-L1 expression in LCSLC and the downstream molecular mechanism of PD-L1 regulating stem cell characteristics, also tumor biological functions. Methods: HepG2 was cultured by sphere-formating method to obtain LCSLC. The expressions of CD133 and other stemness markers were detected by flow cytometry, western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the expressions of stemness markers and PD-L1. The biological functions of the LCSLC were tested by cell function assays, to confirm that the LCSLC has the characteristics of tumor stem cells. LCSLC was treated with cell signaling pathway inhibitors to identify relevant upstream signaling pathways mediating PD-L1 expression changes. The expression of PD-L1 in LCSLC was down regulated by small interfering RNA (siRNA), the expression of stem cell markers, tumor biological functions of LCSLC, and the changes of cell signaling pathways were detected. Results: Compared with HepG2 cells, the expression rate of CD133 in LCSLC was upregulated [(92.78±6.91)% and (1.40±1.77)%, P<0.001], the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were also higher than those in HepG2 cells (P<0.05), the number of sphere-formating cells increased on day 7 [(395.30±54.05) and (124.70±19.30), P=0.001], cell migration rate increased [(35.41±6.78)% and (10.89±4.34)%, P=0.006], the number of transmembrane cells increased [(75.77±10.85) and (20.00±7.94), P=0.002], the number of cloned cells increased [(120.00±29.51) and (62.67±16.77), P=0.043]. Cell cycle experiments showed that LCSLC had significantly more cells in the G(0)/G(1) phase than those in HepG2 [(54.89±3.27) and (32.36±1.50), P<0.001]. The tumor formation experiment of mice showed that the weight of transplanted tumor in LCSLC group was (1.32±0.17)g, the volume is (1 779.0±200.2) mm(3), were higher than those of HepG2 cell [(0.31±0.06)g and (645.6±154.9)mm(3), P<0.001]. The expression level of PD-L1 protein in LCSLC was 1.88±0.52 and mRNA expression level was 2.53±0.62, both of which were higher than those of HepG2 cells (P<0.05). The expression levels of phosphorylation signal transduction and transcription activation factor 3 (p-STAT3) and p-Akt in LCSLC were higher than those in HepG2 cells (P<0.05). After the expression of p-STAT3 and p-Akt was down-regulated by inhibitor treatment, the expression of PD-L1 was also down-regulated (P<0.05). In contrast, the expression level of phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in LCSLC was lower than that in HepG2 cells (P<0.01), there was no significant change in PD-L1 expression after down-regulated by inhibitor treatment (P>0.05). After the expression of PD-L1 was knockdown by siRNA, the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were decreased compared with those of siRNA-negative control (NC) group (P<0.05). The number of sphere-formating cells decreased [(45.33±12.01) and (282.00±29.21), P<0.001], the cell migration rate was lower than that in siRNA-NC group [(20.86±2.74)% and (46.73±15.43)%, P=0.046], the number of transmembrane cells decreased [(39.67±1.53) and (102.70±11.59), P=0.001], the number of cloned cells decreased [(57.67±14.57) and (120.70±15.04), P=0.007], the number of cells in G(0)/G(1) phase decreased [(37.68±2.51) and (57.27±0.92), P<0.001], the number of cells in S phase was more than that in siRNA-NC group [(30.78±0.52) and (15.52±0.83), P<0.001]. Tumor formation in mice showed that the tumor weight of shRNA-PD-L1 group was (0.47±0.12)g, the volume is (761.3±221.4)mm(3), were lower than those of shRNA-NC group [(1.57±0.45)g and (1 829.0±218.3)mm(3), P<0.001]. Meanwhile, the expression levels of p-STAT3 and p-Akt in siRNA-PD-L1 group were decreased (P<0.05), while the expression levels of p-ERK1/2 and ß-catenin did not change significantly (P>0.05). Conclusion: Elevated PD-L1 expression in CD133(+) LCSLC is crucial to maintain stemness and promotes the tumor biological function of LCSLC.

[Association between maternal plasma one-carbon biomarkers during pregnancy and fetal growth in twin pregnancies].

Objective: To characterize the relationship between the levels of plasma methyl donor and related metabolites (including choline, betaine, methionine, dimethylglycine and homocysteine) and fetal growth in twin pregnancies. Methods: A hospital-based cohort study was used to collect clinical data of 92 pregnant women with twin pregnancies and their fetuses who were admitted to Peking University Third Hospital from March 2017 to January 2018. Fasting blood was collected from the pregnant women with twin pregnancies (median gestational age: 18.9 weeks). The levels of methyl donors and related metabolites in plasma were quantitatively analyzed by high-performance liquid chromatography combined with mass spectrometry. The generalized estimation equation was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and neonatal outcomes of twins, and the generalized additive mixed model was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and fetal growth ultrasound indicators. Results: (1) General clinical data: of the 92 women with twin pregnancies, 66 cases (72%) were dichorionic diamniotic (DCDA) twin pregnancies, and 26 cases (28%) were monochorionic diamniotic (MCDA) twin pregnancies. The comparison of the levels of five plasma methyl donors and related metabolites in twin pregnancies with different basic characteristics showed that the median levels of plasma choline and betaine in pregnant women ≥35 years old were higher than those in pregnant women <35 years old, and the differences were statistically significant (all P<0.05). (2) Correlation between plasma methyl donor and related metabolites levels and neonatal growth indicators: after adjusting for confounding factors, plasma homocysteine level in pregnant women with twins was significantly negatively correlated with neonatal birth weight (ß=-47.9, 95%CI:-94.3- -1.6; P=0.043). Elevated methionine level was significantly associated with decreased risks of small for gestational age infants (SGA; OR=0.5, 95%CI: 0.3-0.9; P=0.021) and low birth weight infants (OR=0.6, 95%CI: 0.4-0.9; P=0.020). Increased homocysteine level was associated with increased risks of SGA (OR=1.5, 95%CI: 1.0-2.2; P=0.029) and inconsistent growth in twin fetuses (OR=1.9, 95%CI: 1.0-3.7; P=0.049). (3) Correlation between the levels of plasma methyl donors and related metabolites and intrauterine growth indicators of twins pregnancies: for every 1 standard deviation increase in plasma choline level in pregnant women with twin pregnancies, fetal head circumference, abdominal circumference, femoral length and estimated fetal weight in the second trimester increased by 1.9 mm, 2.6 mm, 0.5 mm and 20.1 g, respectively, and biparietal diameter, abdominal circumference and estimated fetal weight increased by 0.7 mm, 3.0 mm and 38.4 g in the third trimester, respectively, and the differences were statistically significant (all P<0.05). (4) Relationship between plasma methyl donor and related metabolites levels in pregnant women with different chorionicity and neonatal birth weight and length: the negative correlation between plasma homocysteine level and neonatal birth weight was mainly found in DCDA twin pregnancy (ß=-65.9, 95%CI:-110.6- -21.1; P=0.004). The levels of choline, betaine and dimethylglycine in plasma of MCDA twin pregnancy were significantly correlated with the birth weight and length of newborns (all P<0.05). Conclusion: Homocysteine level is associated with low birth weight in twins, methionine is associated with decreased risk of SGA, and choline is associated with fetal growth in the second and third trimesters of pregnancy.

[Study on formulation and revision of standard limit for formaldehyde in the "Standards for indoor air quality（GB/T 18883-2022）" in China].

Formaldehyde, as an important pollutant in indoor air, has always been of great concern. In the newly issued "Standards for indoor air quality (GB/T 18883-2022)", the standard limit of formaldehyde has been restricted to 0.08 mg/m3. In order to better promote the implementation and application of this new standard, this study reviewed and interpreted the relevant technical content for determining the standard limit, including the indoor concentration and human exposure levels of formaldehyde, the health effects of formaldehyde, and the derivation of safety reference values. It also proposed prospect for the future development and revision of quality standards for formaldehyde in indoor air.

[Research and reflection on the diversified method system of multi-stages and multi-scenarios surveillance and early warning of infectious diseases].

With the outbreak of infectious diseases, more and more attention has been paid to surveillance and early warning work. Timely and accurate monitoring data is the basis of infectious diseases prevention and control. Effective early warning methods for infectious diseases can improve the timeliness and sensitivity of early warning work. This paper briefly introduces the intelligent early warning model of infectious diseases, summarizes the emerging surveillance and early warning methods of infectious diseases, and seeks the possibility of diversified surveillance and early warning in different epidemic stages and different outbreak scenarios of infectious diseases. This paper puts forward the idea of constructing a diversified method system of infectious diseases surveillance and early warning based on multi-stages and multi-scenarios and discusses the future development trend of infectious diseases surveillance and early warning, in order to provide reference for improving the construction level of infectious diseases surveillance and early warning system in China.

[Clinicopathological features of patients with RET fusion-positive non-small cell lung cancer].

Objective: To investigate the clinicopathological features, treatment and prognosis of patients with RET fusion positive non-small cell lung cancer (NSCLC). Methods: A total of 1 089 NSCLCs were retrieved at Affiliated Hospital of Jiangnan University from August 2018 to April 2020. In all cases, multiple gene fusion detection kits (fluorescent PCR method) were used to detect the gene status of RET, EGFR, ALK, ROS1, KRAS, BRAF and HER2; and immunohistochemical method was used to detect the expression of PD-L1 and mismatch repair related proteins. The correlation between RET-fusion and patients' age, gender, smoking history, tumor stage, grade, pathologic type, and PD-L1, mismatch repair related protein expression was analyzed. Results: There were 22 cases (2.02%) detected with RET fusion-positive in 1 089 NSCLC patients, in which 11 males and 11 females; and the median age was 63.5 years. There were 20 adenocarcinomas, including 11 acinar predominant adenocarcinoma (APA), five solid predominant adenocarcinoma (SPA) and four lepidic predominant adenocarcinoma (LPA); There were one case each of squamous cell carcinoma (non-keratinizing type) and sarcomatoid carcinoma (pleomorphic carcinoma). There were 6 and 16 patients with RET fusion-positive who were in stage Ⅰ-Ⅱ and Ⅲ-Ⅳ respectively, and 16 cases with lymph node metastasis, 11 cases with distant metastasis. Among RET fusion-positive cases, one was detected with HER2 co-mutation. The tumor proportion score of PD-L1≥1% in patients with RET fusion positive lung cancer was 54.5% (12/22). Defects in mismatch repair protein expression were not found in patients with RET fusion positive NSCLC. Four patients with RET fusions positive (two cases of APA and two cases of SPA) received pratinib-targeted therapy, and two showed benefits from this targeted therapy. Conclusions: The histological subtypes of RET fusions positive NSCLC are more likely to be APA or SPA. RET fusion-positive NSCLC patients are associated with advanced clinical stage, lymph node metastases, and they may benefit from targeted therapy with RET-specific inhibitors.

[Clinical research progress of selective laser trabeculoplasty].

Selective Laser Trabeculoplasty (SLT) is an effective laser treatment modality for controlling intraocular pressure (IOP) in open-angle glaucoma (OAG). Compared to other laser techniques targeting the trabecular meshwork, SLT has demonstrated favorable efficacy and safety. This article aims to summarize the application and advancements of SLT in Chinese OAG patients, focusing on its effectiveness in reducing IOP and postoperative adverse reactions that may influence OAG. By examining SLT from these perspectives, we aim to provide insights for the standardized promotion of SLT and enhancement of clinical glaucoma management in China.

[One case of adult-onset dystonia presenting with chronic progressive external ophthalmoplegia].

We report a case of adult-onset dystonia presenting with chronic progressive external ophthalmoplegia. The patient had ptosis in both eyes, particularly the left eye, for no obvious reason since the age of 10, which was progressively aggravated. The clinical diagnosis was chronic progressive external ophthalmoplegia. However, whole gene sequencing revealed the mitochondrial A3796G missense mutation, so the patient was clearly diagnosed as adult-onset dystonia and given treatment to reduce blood glucose and improve muscle metabolism. The A3796G mutation in the ND1 subunit of the mitochondrial complex leading to ophthalmoplegia is relatively rare, requiring a combination with genetic testing for confirmation of diagnosis.

[Long-term outcome of transcatheter repair of paravalvular leak post surgical mitral valve replacement].

Objective: To explore the long-term clinical efficacy of transcatheter repair of mitral paravalvular leak (PVL) post surgical mitral valve replacement. Methods: This study is a retrospective study. Patients who completed transcatheter repair of paravalvular leak after surgical mitral valve replacement at Shanghai Chest Hospital, Shanghai Jiaotong University School of Medicine from March 2010 to December 2018 were included. Technical success was defined as the occluder being stably implanted in the paravalvular leak site without affecting the function of the mitral valve and surrounding tissues; and there were no intervention-related complications, such as new hemolysis or aggravated hemolysis, and echocardiography confirmed mitral paravalvular regurgitation reduced by more than 1 grade. Patients were followed up at 30 days, 1, and 3 years after the intervention. The main endpoints were all-cause death and re-surgery due to interventional failure or serious complications. The occurrence of occluder-mediated hemolysis and chronic renal insufficiency was recorded, and patients were monitored with echocardiography during follow up. Results: A total of 75 patients were included, aged (54.3±22.9) years old, and 38 patients were males. All patients had decreased cardiac function and/or hemolysis before intervention. Procedural success was achieved in 54 patients (72.0%). Incidence of device-mediated hemolysis was 18.7% (14/75). During the follow-up period, all-cause death occurred in 7 patients (9.3%), and 3 were cardiac deaths.The 3-year event-free survival rate was 81.3% (61/75). The need for cardiac surgery was 9.3% (7/75): 3 cases due to severe device-mediated hemolysis, 2 cases due to prosthetic valve failure and 2 cases due to moderate to severe residual regurgitation. The echocardiography follow-up results showed that the position of the occluder was stable, there was no impact on the artificial valve function and surrounding structures, and the residual regurgitation was stable without progressive increase in event-free patients. Compared with pre-intervention, the left ventricular end systolic diameter ((33.9±7.4)mm vs. (38.3±8.9) mm, P=0.036), end diastolic diameter ((53.7±8.3) mm vs. (58.4±9.1) mm, P=0.045) and left atrial diameter (59.3 (44.5, 90.7) mm vs. 64.3 (44.8, 96.6) mm, P=0.049) were significantly reduced, pulmonary artery systolic pressure was also significantly decreased ((36.5±15.8) mmHg vs. (46.3±14.9) mmHg, P=0.022, 1 mmHg=0.133 kPa). There was no significant difference between 3 years and 1 year after transcatheter repair of mitral paravalvular leak post surgical mitral valve replacement (all P>0.05). Conclusion: Transcatheter repair of mitral paravalvular leak post surgical mitral valve replacement is an effective treatment option in selective patients.

Features and clinical significance of tertiary lymphoid structure in cutaneous squamous cell carcinoma.

BACKGROUND: Tertiary lymphoid structure (TLS) plays an important role in antitumour immunity, largely reflecting the prognosis. However, its clinical implication in cutaneous squamous cell carcinoma (cSCC) remains unknown. OBJECTIVES: To explore the features of TLS in cSCC and its association with clinicopathological characteristics. METHODS: Two independent RNA-seq data of cSCC were used to investigate the tumour immune microenvironment, as well as TLS-related chemokines and cytokines. The density and location of TLSs were assessed in a total of 82 cSCC patients, and the clinicopathologic association was examined. RESULTS: Bioinformatics analysis showed that a large amount of immune cell infiltration and significant up-regulation of TLS-related chemokines were observed in cSCC. Histologically, TLSs appeared as highly organized structures in 72 (87.8%) cases with different levels of density and maturation, among which 14 cases were in low-density group and 58 cases were in high-density group. Clinically, the presence of TLS was prominently associated with better degree of histopathological grades and higher level of sun exposure. Furthermore, the presence of intratumoral TLS was associated with lower lymphovascular invasion. CONCLUSIONS: TLS is highly organized in cSCC, and the presence of TLS is a positive prognostic factor for cSCC, which will provide a theoretical basis for the future diagnostic and therapeutic value in cSCC.

Physical activity and sedentary behaviour among children and adolescents with intellectual disabilities during the COVID-19 lockdown in China.

BACKGROUND: In the wake of the COVID-19 pandemic, preliminary research has reported a significant decline in physical activity (PA) and an increase in sedentary behaviour (SB) among typically developed children and adolescents. Limited research has looked at the current situation of PA and SB during this pandemic among children and adolescents with intellectual disabilities (ID). This study investigated the situations about PA and SB among school-aged children and adolescents with ID on China's mainland during the COVID-19 outbreak. METHODS: In total, 837 parents of children and adolescents (ages 6-18 years) with ID from 15 special education schools of Shandong Province in China were recruited through convenience sampling in the study. Parents reported PA and SB among children and adolescents with ID through the Children's Leisure Activities Study Survey-Chinese version (CLASS-C) online questionnaires. RESULTS: From parents' reports, Chinese children and adolescents with ID during the COVID-19 pandemic participated in approximately 10 min of moderate-to-vigorous physical activity, and engaged in approximately 530 min of SB every day. Meanwhile, only 17.4% of children and adolescents with ID were able to achieve the recommendation of 60 min of daily moderate-to-vigorous physical activity and 76.1% of children and adolescents with ID spent more than 2 h on SB per day. Additionally, the problems of decrease PA and excessive SB were more prominent in older adolescents with ID compared with younger children with ID. CONCLUSION: In China, the low level of PA and high level of SB is particularly evident in children and adolescents with ID during the outbreak of COVID-19. The great majority of children and adolescents with ID did not meet the recommended amount of PA while undergoing excessive SB under the long-term home quarantine environment. Therefore, immediate attention and great effort should be made to deal with this severe situation among this vulnerable population in the mainland of China.

[Influence of Helicobacter pylori infection and its eradication treatment on small intestinal bacterial overgrowth].

Objectives: To analyze the effects of Helicobacter pylori (H. pylori) infection and eradication therapy on small intestinal bacterial overgrowth (SIBO). Methods: From September to December 2021, patients with abdominal symptoms who received 13C urea breath test at the Department of Gastroenterology in Peking University First Hospital were enrolled.13C urea breath test was used to detect H. pylori infection and patients were divided into H. pylori positive and negative groups accordingly. Lactulose hydrogen methane breath test was performed to determine SIBO. H. pylori positive patients were treated with quadruple therapy including amoxicillin, metronidazole, rabeprazole and bismuth potassium citrate. 13C urea breath test and lactulose hydrogen methane breath test were reexamined 6 weeks after the treatment. Results: A total of 102 patients (49 males and 53 females) were enrolled, with a mean age of (42.1±9.9) years. Among them, 49 patients were H. pylori negative and 53 were H. pylori positive. Moreover, 47 patients were SIBO positive and 55 were SIBO negative. There was no significant difference in age, sex, body mass index, abdominal symptoms and the diagnosis of chronic atrophic gastritis between H. pylori positive and negative patients at the enrollment (all P>0.05). The detection rate of SIBO in H. pylori infected patients was higher than that in uninfected patients, and the difference was statistically significant (60.4% vs 30.6%, P=0.003). Patients with SIBO had significantly more frequent abdominal distension (36.2% vs 10.9%, P=0.002) and constipation (27.7% vs 1.8%, P<0.001) than patients without SIBO. The rate of SIBO after H. pylori eradication treatment was significantly lower than that before treatment (20.8% vs 60.4%, P<0.001). The remission rate of SIBO after eradication therapy was 66.7% (20/30). Besides, patients had obvious improvement of constipation (6.0% vs 18.9%, P=0.008), and the incidence of other abdominal symptoms decreased to various degrees including diarrhea, abdominal pain, abdominal distention and poor appetite. Conclusion: H. pylori infection increases the risk of SIBO, and the quadruple regimen containing amoxicillin and metronidazole has a therapeutic effect for patients with H.pylori infection and concomitant SIBO.

[Efficacy of dienogest versus gonadotropin-releasing hormone agonist combined with dienogest sequential therapy in the treatment of adenomyosis].

Objective: To investigate the efficacy and safety of dienogest (DNG) alone and gonadotropin-releasing hormone agonist (GnRH-a) combined with DNG sequential treatment to adenomyosis. Methods: The clinical data of 110 patients with adenomyosis attending the First Affiliated Hospital of Nanjing Medical University from December 2019 to March 2022 were retrospectively analyzed, including 40 patients treated with DNG (2 mg/day) alone (DNG group) and 70 patients treated with sequential DNG (2 mg/day) after 3-6 injections of GnRH-a (GnRH-a+DNG group). The clinical data before and after treatment were compared between the two groups. Results: (1) The dysmenorrhea visual analogue scale (VAS) scores, cancer antigen 125 (CA125) and cancer antigen 19-9 (CA19-9) levels at different time periods after treatment were significantly lower than before treatment in both groups (median before treatment: DNG group 70.0 mm, 68.55 kU/L, 22.45 kU/L respectively, GnRH-a+DNG group 80.0 mm, 151.50 kU/L, 20.44 kU/L respectively; all P<0.001). (2) The hemoglobin (Hb) levels of patients in both groups at different time periods after treatment were significantly higher than those before treatment (median: DNG group 102.00 g/L, GnRH-a+DNG group 94.00 g/L; all P<0.001). (3) Treatment with DNG alone did not have a significant effect on uterine volume in patients of DNG group (P>0.05), and uterine volume decreased significantly in the 15th-24th months of GnRH-a+DNG group compared with that before treatment (median: 167.76 vs 227.77 cm3; P<0.05). (4) There were no significant differences in hepatic and renal function and coagulation indexes between the two groups before and after treatment (all P>0.05), and no significant abnormal lesions were observed in breast tissue during the follow-up period. (5) The incidence of amenorrhea of GnRH-a+DNG group was higher than that of DNG group, and the incidences of irregular spotting bleeding and breakthrough hemorrhage were lower than those in DNG group. Conclusions: Whether DNG is used alone or in combination with GnRH-a in sequence, it could significantly relieve dysmenorrhea symptoms, improve the level of Hb, reduce the levels of CA125 and CA19-9 in patients with adenomyosis, with no adverse effects on coagulation and hepatic or renal function. GnRH-a sequential DNG therapy is superior to DNG alone in improving uterine bleeding patterns and controlling the growth of uterine volume in patients with adenomyosis.

[Changes of coagulation function in patients with adenomyosis].

Objective: To explore the different coagulation state in patients with adenomyosis and its clinical significance. Methods: Clinical data of the patients admitted to the First Affiliated Hospital of Nanjing Medical University from January 2017 to December 2021 were retrospectively analyzed. (1) Differential coagulation state between 25 healthy women and 25 patients with adenomyosis were compared during menstrual and non-menstrual periods. (2) The coagulation indexes of 145 patients with adenomyosis (observation group 1) and 129 patients with cervical intraepithelial neoplasia grade Ⅲ (control group 1) who underwent hysterectomy in non-menstrual period were compared. (3) The coagulation indexes of 154 patients with adenomyosis (observation group 2) and 147 women without myometrial lesions (control group 2) who underwent endometrial curettage during uterine bleeding period were compared. (4) Correlations of coagulation index with cancer antigen 125 (CA125), cancer antigen 19-9 (CA19-9) and uterine volume in patients with adenomyosis were analyzed. Results: (1) The coagulation state of each health women during the menstrual and non-menstrual period showed no significant differences (all P>0.05). For the 25 patients with adenomyosis, fibrinogen [FIB; 2.61 g/L(2.50-3.10 g/L)] and D-dimer [0.60 mg/L (0.40-1.00 mg/L)] in the menstrual period were significantly higher than those in the non-menstrual period [2.25 g/L (1.90-2.70 g/L) and 0.27 mg/L (0.20-0.40 mg/L), respectively; both P<0.01], while thrombin time [TT; 16.70 s (16.10-17.40 s)] in the menstrual period was significantly lower than that in the non-menstrual period [17.95 s (17.20-18.40 s); P<0.01]. (2) In the non-bleeding period, D-dimer [0.26 mg/L (0.20-0.40 mg/L)] and platelet count [257.0×109/L (212.0×109/L-308.5×109/L)] of observation group 1 were significantly higher than those of control group 1 (all P<0.01). Besides, FIB (r=0.237, P=0.004) and D-dimer (r=0.373, P<0.001) were positively correlated with CA125, while prothrombin time (PT; r=-0.208, P=0.012) and internationalized normalized ratio of plasma prothrombin time (PT-INR; r=-0.201, P=0.015) were negatively correlated with CA19-9. (3) In the bleeding period, PT [10.70 s (10.10-11.20 s)] and PT-INR [0.93 (0.90-1.00)] of observation group 2 were significantly lower than those of control group 2 (all P<0.01), while D-dimer [0.41 mg/L (0.20-0.80 mg/L)] was significantly higher than that in the control group 2 (P<0.001). Furthermore, FIB (r=0.252, P=0.038) and D-dimer (r=0.321, P=0.008) were positively correlated with uterine volume, while activated partial thromboplastin time (APTT; r=-0.190, P=0.018) and TT (r=-0.304, P=0.012) were negatively correlated with uterine volume. (4) During non-menstrual period and uterine bleeding period, APTT and TT in patients of observation group 1 and 2 combined with anemia were significantly lower than those of non-anemia patients (all P<0.05). Conclusion: Patients with adenomyosis have a tendency to hypercoagulability in both the uterine bleeding and non-bleeding periods, which may be related to enlarged uterine volume, increased serum CA125 and anemia.

[Evaluation of safety and immunogenicity of hepatitis E vaccine in maintenance hemodialysis patients].

Objective: To evaluate the safety and immunogenicity of hepatitis E vaccine(HEV)in Maintenance hemodialysis(MHD)patients. Methods: Based on an open-labeled controlled trial, from May 2016 to March 2018, 35 eligible MHD patients were recruited in the Hemodialysis Center of Zhongshan Hospital Affiliated to Xiamen University as the experimental group, and 70 MHD patients with matched age, gender and underlying diseases as the control group. The experimental group received HEV at 0, 1 and 6 months according to the standard vaccination procedures, while the control group received routine diagnosis and treatment without vaccine and placebo injection to observe the safety and immunogenicity of the vaccine. The safety of vaccine in MHD population was evaluated by the incidence of adverse reactions/events in the experimental and control groups. The immunogenicity of HEV in MHD patients was evaluated by comparing the data from the phase Ⅲ clinical trial. Results: The overall incidence of adverse reactions/events was 17.1% (18/105), and there were no grade 3-4 adverse reactions/events related to vaccination. In the experimental group, the incidence of local adverse reactions/events was 20.0% (7/35), and the incidence of systemic adverse reactions/events was 17.1% (6/35).There was no significant difference in the incidence of systemic adverse reactions/events between the experimental group and the control group (P>0.05). There were 23 patients receiving 3 doses with the standard schedule. The positive rate of HEV-IgG antibody was 100% and the GMC was 14.47(95%CI:13.14-15.80) WU/ml, which showed no significant difference compared with the 46 patients in Phase Ⅲ clinical trial (t=-1.04, P>0.05). Conclusion: Recombinant HEV has good safety and immunogenicity in MHD patients.

[Identification and analysis of 2 Corynebacterium diphtheria strains in Guangdong Province].

Objective: To identify and analyze two strains of C. diphtheriae in Guangdong Province by combining whole genome sequencing with traditional detection methods. Methods: The C. diphtheriae was isolated from Guangzhou in 2010 and Zhuhai in 2020 respectively. Isolates were identified by API Coryne strips and MALDI-TOF-MS. Genomic DNA was sequenced by using Illumina. The assembly was performed for each strain using CLC software. J Species WS online tool was used for average nucleoside homology identification, then narKGHIJ and tox gene were detected by NCBI online analysis tool BLSATN. MEGA-X was used to build a wgSNP phylogenetic tree. Results: GD-Guangzhou-2010 was Belfanti and GD-Zuhai-2020 was Gravis. ANIb between GD-Guangzhou-2010 and C. belfantii was 99.61%. ANI between GD-Zhuhai-2020 and C. diphtheriae was 97.64%. BLASTN results showed that the nitrate reduction gene narKGHIJ and tox gene of GD-Guangzhou-2010 was negative, while GD-Zhuhai-2020 nitrate reduction gene narKGHIJ was positive. There were two obvious clades in wgSNP phylogenetic tree. The first clades included all Mitis and Gravis types strains as well as GD-Zhuhai-2020. The second clades contained all isolates of C.belfantii, C.diphtheriae subsp. lausannense and GD-guangzhou-2010. Conclusion: Two non-toxic C. diphtheriae strains are successfully isolated and identified. The phylogenetic tree suggests that GD-Guangzhou-2010 and GD-Zhuhai-2020 are located in two different evolutionary branches.

Two polyurethanases PueA and PueB are major extracellular lipases partly secreted by the mediation of their cognate ABC exporter AprDEF in Pseudomonas protegens Pf-5.

Two polyurethanases PueA and PueB from Pseudomonas protegens Pf-5 have been reported to have hydrolytic activity against synthetic p-nitrophenyl palmitate of lipase substrate, and PueA may play a more effective role in this activity. However, it is still unknown whether PueA and PueB play similar parts in the lipase activity against natural acylglycerols and achieve the extracellular secretion via their cognate ABC exporter AprDEF. In this study, we investigated these questions through the construction of four markerless deletion mutants in Pf5139 (Δupp derivative of Pf-5), two heterologous co-expression strains and their three control strains in lipase-free Escherichia coli BL21(DE3), and detected their lipase activities by the tributyrin plate assay and the liquid culture assay. The results showed that PueA and PueB, classified as subfamily I.3 lipases, are major extracellular lipases involved in the uptake of oil in Pf-5, and PueA plays a leading role in extracellular lipase activity. In addition, the extracellular secretion of PueA and PueB can be partly mediated via AprDEF in Pf-5 and BL21(DE3). Finally, PueA and PueB are also able to achieve the extracellular secretion without the assistance of AprDEF in Pf-5 and BL21(DE3).

Prevalence of overweight and obesity in children and adolescents with intellectual disabilities in China.

BACKGROUND: Overweight and obesity in children and adolescents has become a worldwide epidemic. There are several studies that have concentrated on the prevalence rate of children and adolescents with intellectual disabilities (ID), whereas data on such a population on the mainland of China remain unclear. The purpose of this study was to investigate the prevalence rate of overweight and obesity among school-aged children and adolescents with ID on China's mainland. METHODS: This study employed a cross-sectional design to examine the body weight status of 1873 children and adolescents (ages 6-18 years old) with ID in 35 special education schools. Body mass index was calculated, and the concepts of overweight and obesity were defined according to the standard of the Working Group for Obesity in China. RESULTS: Data indicated that 18.2% (95% CI: 16.5%-20.0%) of children and adolescents with ID were overweight and 14.4% (95% CI: 12.8%-16.0%) were obese. Boys with ID were more likely to be overweight than girls with ID (OR = 1.48[95% CI: 1.13-1.94], P < 0.05). Children and adolescents with Down syndrome or autism spectrum disorder had a trend to be classified as overweight (OR = 1.76[95% CI: 1.22-2.54], P < 0.05; OR = 1.57[95% CI: 1.17-2.09], P < 0.05, respectively) or obesity (OR = 1.82[95% CI: 1.23-2.69], P < 0.05; OR = 1.40 [95% CI: 1.02-1.93], P < 0.05, respectively) compared with those with ID without these conditions. Moreover, children and teenagers with ID living in urban areas had a predisposition to be overweight (OR = 2.16[95% CI: 1.14-4.09], P < 0.05) or obese (OR = 3.25[95% CI: 1.41-7.50], P < 0.05) relative to those who lived in rural areas. CONCLUSION: Results indicated that in China, the prevalence rate of overweight and obesity among school-aged children and adolescents with ID was remarkably high. Therefore, future research should make every effort to focus on reducing and preventing overweight and obesity of this population in China.

[Establishment of model to predict lateral neck recurrence of central lymph node metastasis in papillary thyroid carcinoma].

Objective: To explore the risk factors for lateral neck recurrence of central lymph node metastasis (CLMN) in papillary thyroid cancer (PTC), and to construct a model to predict the recurrence. Methods: The records of 245 consecutive PTC patients with CLMN underwent surgical treatment from 1996 to 2009 in our department were retrospectively reviewed. The threshold value of CLNM number is determined by ROC curve. The risk factors for lateral neck recurrence were determined by using Cox regression model. The identified risk factors were incorporated into a nomogram model to predict the risk of lateral neck recurrence. Results: A total of 245 patients were enrolled in the study, among them, 32 cases occurred lateral neck lymph node recurrence and 4 cases were dead of thyroid carcinoma. Multivariate analysis revealed that primary tumor size, extrathyroidal extension, the number of metastatic CLNM >3 were independent risk factors of lateral neck recurrence (P<0.05), lateral neck recurrence was a risk factor of disease-free survival(P<0.05). The nomogram model of predicting the lateral neck recurrence was further established based on the above 3 independent risk factors, the area under the receiver operating characteristic (ROC) curve of which was 0.790. Conclusions: The nomogram model based on the independent risk factors of LN recurrence can be helpful to screen the papillary thyroid carcinoma patients with high risk of lateral neck recurrence, and provide more guidance for clinical treatment.

[Prediction and investigation of the potential risk factors for the upper mediastinal metastasis of papillary thyroid carcinoma].

Objective: To predict and investigate the potential risk factors for the upper mediastinal metastasis of papillary thyroid carcinoma (PTC). Methods: This study was a prospective cohort study. The admission criteria were patients with untreated thyroid cancer diagnosed in Cancer Hospital, Chinese Academy of Medical Sciences from December 2013 to December 2015, and positive lymph node (cN1, including cN1a and cN1b) was diagnosed by ultrasound. All patients underwent neck to thorax enhanced Computed Tomography (CT) examination preoperatively. All patients with suspected upper mediastinal lymph node metastasis experienced suspicious regional dissection, and those who had not undergone surgery and whose postoperative pathology was non-papillary thyroid carcinoma were excluded. Kaplan-Meier method was selected for survival analysis and all the factors were analyzed by multivariate Logistic regression. Results: Of the 248 patients, 54 were prompted by postoperative pathology for upper mediastinal lymph node metastasis, 86 cases were phase T1, 94 cases were phase T2, 17 cases were phase T3 and 51 cases were phase T4, 21 cases were N1a phase and 227cases were N1b phase. There was a statistically significant difference in the T-phase and N-phase between the upper mediastinal lymph node metastasis group and no upper mediastinal lymph node metastasis group(P<0.05). Univariate analysis showed that among the preoperative relevant factors, ultrasound tumor length> 2 cm, ultrasound tumor bilaterally, CT double neck lymph node metastasis, increased thyroglobulin (Tg), and increased anti-thyroglobulin antibody (ATG) were all risk factors for upper mediastinal lymph node metastasis(all P<0.05). Among the postoperative factors, bilateral tumor, double neck lymph node metastasis, tumor invasion of the recurrent laryngeal nerve, trachea, esophagus or larynx, T3 staging, T4 staging, total number of metastatic lymph nodes>10, the number of metastatic lymph nodes in level Ⅵ>3 and >6, the proportion of metastatic lymph nodes in level Ⅵ>1/2, the number of metastatic lymph nodes in level Ⅳ> 5 and metastatic proportion >1/3 are risk factors for metastasis of upper mediastinal lymph node(all P<0.05). Multivariate analysis showed that CT indicated double neck lymph node metastasis, increased Tg, increased ATG, the proportion of metastatic lymph nodes in level Ⅵ >1/2, and in level Ⅳ>1/3 are independent risk factors for upper mediastinum lymph node metastasis(all P<0.05). The 5-year recurrence-free survival rates of the upper mediastinal lymph node metastasis group and the no upper mediastinal lymph node metastasis group were 92.3% and 94.8% respectively, and the difference was not statistically significant(P=0.307). Conclusions: For preoperative ultrasound considering the presence of lymph node metastases, enhanced neck to thorax CT should be performed routinely. When bilateral cervical lymph node metastasis is determined by CT, or endocrine tests suggest abnormally increased antibodies, attention should be paid to the upper mediastinal lymph nodes metastasis. In the course of neck dissection, if more lymph node metastases in level Ⅵ and level Ⅳ were detected, surgeons should be vigilant of the upper mediastinal metastasis. The prognosis of patients underwent complete mediastinal dissection is not significantly different from that of patients without mediastinal metastasis.