Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.

Objective:To evaluate the value of 18F-FDG PET/CT for preoperative localization of epileptogenic foci in refractory epilepsy patients with negative MRI. Methods:Clinical data (550 lobes) of 55 epilepsy patients (38 males, 17 females, age (20.0±8.1) years) with negative MRI who underwent preoperative 18F-FDG PET/CT-MRI between January 2014 and June 2020 at the First Affiliated Hospital of Jinan University were retrospectively analyzed. The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of 18F-FDG PET/CT, video electroencephalogram (VEEG), PET/CT+ VEEG and PET/CT-VEEG for localizing epileptogenic foci were calculated using stereoelectroencephalography (SEEG) and the outcomes of at least 1 year of postoperative follow-up as reference standards. χ2 test was used to compare the efficiencies of different examination modalities for unilobar, multilobar and all patients. Results:The correct lateralization rate of epileptogenic foci was 92.6%(25/27) using PET/CT. The sensitivity, specificity, accuracy, PPV and NPV of PET/CT for localization of epileptogenic foci were 65.1%(54/83), 77.9%(364/467), 76.0%(418/550), 34.4%(54/157) and 92.6%(364/393), respectively. The sensitivities of PET/CT-VEEG for localization of epileptogenic foci in all patients and patients with multilobar epilepsy were higher than those of VEEG alone (75.9%(63/83) vs 45.8%(38/83), 68.6%(35/51) vs 31.4%(16/51); χ2 values: 15.80, 14.16, both P<0.001). The specificities of PET/CT+ VEEG for localization of epileptogenic foci in all patients and patients with unilobar epilepsy were higher than those of VEEG alone (97.6%(456/467) vs 94.6%(442/467), 97.9%(282/288) vs 94.1%(271/288); χ2 values: 5.66, 5.48; P values: 0.017, 0.019). The sensitivity of PET/CT-VEEG (PET/CT and VEEG concordance) for localization of epileptogenic foci was higher than that of PET/CT+ VEEG (PET/CT and VEEG discordance) (8/9 vs 28.4%(21/74); χ2=10.40, P=0.001), and its specificity and accuracy were higher than those of PET/CT-VEEG (PET/CT and VEEG discordance) (93.4%(57/61) vs 71.7%(291/406), 92.9%(65/70) vs 72.1%(346/480); χ2 values: 13.23, 13.96; both P<0.001). Conclusions:18F-FDG PET/CT can localize and lateralize epileptogenic foci in patients with negative MRI. The combination of 18F-FDG PET/CT and VEEG improves the sensitivity, specificity, and accuracy for epileptogenic foci detection. 18F-FDG PET/CT is more accurate in detecting epileptogenic foci when it is concordant with VEEG.

Objective:To investigate the impact of the deep-learning-based CT fractional flow reserve (CT-FFR) on clinical decision-making and long-term prognosis in patients with obstructive coronary heart disease.Methods:In this single-center retrospective cohort study, consecutive patients with obstructive coronary heart disease (with at least one stenosis≥50%) on their first coronary computed tomography angiography (CCTA) in Beijing Anzhen Hospital from February 2017 to July 2018 were included. Baseline clinical and CT characteristics were collected. Deep-learning-based CT-FFR and Leiden CCTA risk score were calculated. All patients enrolled were followed up for at least 5 years. The study endpoint was major adverse cardiovascular events (MACE), defined as the composite of cardiac death, nonfatal myocardial infarction, unstable angina requiring hospitalization, and unplanned revascularization. Receiver operating characteristic (ROC) curves were drawn to define the optimal cut-off point of the Leiden score in predicting the 5-year MACE, and survival analysis and Cox regression were performed to explore the related factors of MACE.Results:A total of 622 patients, aged 61 (54, 66) years, with 407 (65.4%) males were included. Diagnostic coronary angiography was performed in 78 patients after their baseline CCTA, with 34 (43.6%) patients had CT-FFR>0.80. During a follow-up time of 2 181 (2 093, 2 355) days, 155 patients (24.9%) suffered from MACE. ROC derived optimal cut-off point of Leiden score for predicting MACE was 15.48. Survival analysis found that male patients, Leiden risk score>15 and CT-FFR≤0.80 had worse prognosis. Multivariate Cox regression analysis identified CT-FFR≤0.80 as an robust and independent predictor of MACE ( HR=4.98, 95% CI 3.15-7.86, P<0.001). Conclusion:Deep-learning-based CT-FFR aids in clinical decision-making and the evaluation of long-term prognosis in patients with obstructive coronary heart disease.

Objective To investigate the safety of using a self-made new type non-inflatable retractor for transoral endoscopic thyroidectomy via submental and vestibular approach(TOETSMVA).Methods A retrospective analysis was conducted on the surgical records of 119 cases of unilateral thyroid lobe tumor from January 2021 to June 2022.Among them,37 cases underwent non-inflatable TOETSMVA by using a new type non-inflatable retractor(non-inflatable group),37 cases underwent traditional inflatable TOETSMVA(inflatable group),and 45 cases underwent traditional open anterior cervical thyroid surgery(open group).The differences in surgical indicators,postoperative complications,and patient satisfaction with incision were compared among the three groups.Results Compared with the open group,the non-inflatable group had longer operation time[(131.0±20.1)min vs.(81.1±15.7)min,P=0.000],but less intraoperative blood loss[19(15,27)ml vs.30(25,37)ml,P=0.000],lower pain score on the first day after surgery[(4.8±2.1)points vs.(6.4±1.9)points,P=0.000],and higher patient's satisfaction with the incision(the number of cases of very dissatisfied,dissatisfied,satisfied,comparatively satisfied,and very satisfied were 0,0,6,16,15 vs.4,3,19,17,2,P=0.000).Compared with the inflatable group,the non-inflatable group had shorter surgical time[(131.0±20.1)min vs.(141.8±22.9)min,P=0.019],and there were no statistically significant differences in intraoperative bleeding volume,pain score on the first day after surgery,and patient satisfaction with the incision(P＞0.05).There were no statistically significant differences in the number of lymph node dissection,total postoperative drainage volume,and parathyroid hormone(PTH)levels on the first day after surgery among the three groups(P＞0.05).Postoperative bleeding occurred in 1 case in the open group,and 1 case of hoarseness and 1 case of subcutaneous ecchymosis of the neck were noted in the non-inflatable group.No tracheoesophageal injury,severe subcutaneous emphysema,hypercapnia,or gas embolism happened among the three groups.Conclusions The new type non-inflatable retractor can effectively maintain space during TOETSMVA surgery.Compared with traditional open anterior cervical thyroid surgery and conventional inflatable TOETSMVA,it has better surgical safety.

Objective:To develope a titanium specimen with good osteogenic activity through fabrication of a composite hydroxyapatite coating on ordered micro-/nanotextured titanium surface.Methods:An ordered micro-/nanotextured structure was prepared on the surface of titanium (the control), and then hydroxyapatite was deposited on the as-prepared ordered micro-/nanotextured structure by alternative loop immersion method. The ordered micro-/nanotextured structures before and after hydroxyapatite deposition were denoted as HA and MN, respectively. Surface morphology was observed using a scanning electron microscope. Bone marrow mesenchymal stem cells (BMMSC) were seeded on the surface of three different materials. Cell morphology was observed with a scanning electron microscope. Cell adhesion and cell proliferation were evaluated using 4', 6-diamidino-2-phenylindole staining and cell counting kit-8 assay, respectively. Extracellular matrix mineralization and the expression levels of osteogenesis-related genes were evaluated by alizarin red staining and real-time quantitative PCR, respectively. Each group has three samples in every experiment.Results:After alternative loop immersing, the MN's original microholes (20 μm in diameter) were retained, and the uniform petal-like hydroxyapatite was deposited on the MN's original titania nanotubes (70 nm in diameter). Compared with the control, BMMSC on MN and HA elongated further and intersected along the micron structure with noticeable pseudopodia and pseudoplates, and the trend was more pronounced especially on HA. The number of early adherent cells on HA was remarkably larger than that on the control and MN at each time point ( P<0.05). On day 1, the A value of cell proliferation on HA was significantly higher than that on the control and MN ( P<0.05). The A value of cell proliferation on HA was significantly lower than that on the control and MN on day 3 ( P<0.05). On day 7, the A value of cell proliferation on HA was significantly lower than that on MN ( P<0.05), but there was no statistically significant difference in the A value of cell proliferation between HA and the control on day 7 ( P>0.05). The Avalue of extracellular matrix mineralization on HA (0.607±0.011) was significantly higher than that on the control and MN (0.268±0.025 and 0.522±0.022, respectively) ( t=-0.25, P<0.001; t=-0.34, P<0.001). The expression levels of bone related genes on HA were significantly higher than those on the control and MN ( P<0.05). Conclusions:HA could promote the BMMSC adhesion and osteogenic differentiation, support BMMSC proliferation, and demonstrate good osteogenic activity.

Background and Aims:The repair of giant incisional hernia is challenging,as closing the significant defect in the abdominal wall can lead to life-threatening complications like abdominal compartment syndrome(ACS).Botulinum toxin type A(BTA)can temporarily relax the abdominal wall muscles,facilitating defect repair,while preoperative progressive pneumoperitoneum(PPP)can increase intra-abdominal volume,reducing intra-abdominal pressure caused by hernia content reintegration.Combining BTA with PPP for the preoperative preparation of giant incisional hernia repair may have a complementary effect.This study was conducted to evaluate the clinical value of combining BTA and PPP in the repair of giant abdominal incisional hernia. Methods:The clinical data of 213 patients with giant abdominal incisional hernia treated at the Sixth Affiliated Hospital of Sun Yat-sen University from December 2015 to December 2019 were retrospectively analyzed.Two weeks after receiving combined BTA and PPP treatment,changes in bilateral abdominal wall muscle,intra-abdominal adhesions,abdominal circumference,abdominal cavity volume,and hernia sac volume ratio were assessed using CT.Intraoperative details,incidence of complications,and postoperative follow-up outcomes were recorded. Results:Following combined BTA and PPP treatment,CT scan showed a significant extension of bilateral lateral abdominal wall muscles towards the midline in all 213 patients,with an average increase of 2.45(1.53-3.29)cm on the left side and 2.54(1.68-3.40)cm on the right side;muscle thickness was reduced by an average of 0.84(0.64-1.00)cm on the left and 0.82(0.62-1.05)cm on the right,the average distance between viscera and the abdominal wall increased to(7.52±1.78)cm,with a mean increase of 6.1(4.2-6.9)cm;the mean increase in abdominal cavity volume was 1 802(1 494.98-2 316.26)mL,and the hernia sac volume ratio decreased by an average of 9％(6％-12％),all changes were statistically significant(P＜0.05).Post-PPP CT scan revealed no abdominal adhesions in 18 patients(8.45％),while 195 patients(91.55％)had varying degrees of adhesions,including 39 cases(18.31％)of sheet adhesions and 156 cases(73.24％)of mixed adhesions.Adhesions mainly consisted of omentum and intestinal tissues in 59.15％of cases.There were 43 cases(20.19％)of grade Ⅰ complications during the BTA-PPP process,including abdominal pain(28 cases),shoulder pain(9 cases),subcutaneous emphysema(6 cases),and dyspnea(3 cases).Dyspnea improved with oxygen therapy,while other complications required no special intervention.All 213 patients successfully underwent laparoscopic incisional hernia repair without conversion to open surgery or organ resection for volume reduction.Fascial closure was achieved in 209 cases(98.12％),with 4 cases(1.88％)having incomplete defect closure.The average time for adhesiolysis was 28(11.00-44.50)min,with a total operative time of 178.0(132.50-255.00)min and an average blood loss of 20(10-30)mL.The median intra-abdominal pressure(IAP)after operation was between 10 mmHg(9.00-12.00 mmHg),Among them,47 cases(22.07％)had IAP exceeding 12 mmHg,and after implementing proactive measures such as diuresis and diachoresis to reduce intra-abdominal contents,the IAP in these patients decreased to below 12 mmHg.No severe complications such as skin flap necrosis or ACS were observed.There were no deaths within postoperative 30 d,and during a follow-up period of 26(16.50-33.00)months,13 cases(6.10％)had surgical site events,including infections in 5 cases(2.35％),seromas in 7 cases(3.29％),and hematoma in 1 case(0.47％),with no hernia recurrence. Conclusion:The combination of BTA and PPP not only aids in identifying abdominal wall adhesion areas,improving preoperative surgical planning and enhancing surgical safety,but also significantly increases abdominal cavity volume and extends lateral abdominal wall muscles,facilitating the closure of giant incisional hernia defects and reducing the incidence of severe postoperative complications like ACS.This approach is worthy of clinical promotion.

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.

OBJECTIVE: To present on a prenatally diagnosed case with complex structural rearrangements of chromosome 8. METHODS: Chromosome karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) were carried out for a fetus with increased nuchal thickness. RESULTS: The karyotype of the amniotic fluid sample showed extra materials on 8p. FISH revealed a centromeric signal at the terminal of 8p with absence of telomeric signal. CMA revealed partial deletion of 8p23.3 [(208049_2256732)×1], partial duplication of 8p23.3p23.2 [(2259519_3016818)×3], and partial duplication of 8q [8q11.1q12.2(45951900_60989083)×3]. CONCLUSION The complex structural rearrangements of chromosome 8 in this case has differed from the commonly seen inv dup del(8p).
Female ; Pregnancy ; Humans ; Chromosomes, Human, Pair 8/genetics* ; In Situ Hybridization, Fluorescence ; Gene Rearrangement ; Prenatal Diagnosis ; Centromere

Sepsis is a life-threatening condition for patients. Biomarkers can be used for the diagnosis, treatment, and prognostic assessment of sepsis. In recent years, new biomarkers for sepsis have been discovered, and more than 250 biomarkers have been identified so far. The complexity of the sepsis process and the increased sensitivity of various detection techniques will lead to the emergence of new biomarkers. However, there is still a lack of specific diagnostic biomarkers and effective therapeutic approaches for sepsis in clinical practice. Therefore, the search for reliable biomarkers and the evaluation of the role of biomarkers in sepsis will undoubtedly aid in clinical decision-making. This article reviews the advances on research of sepsis biomarkers in order to improve understanding of current biomarkers of sepsis, and provide reference for the application of biomarkers in clinical diagnosis, treatment, and prognosis of sepsis.

BACKGROUND: Hypertension is associated with stroke-related mortality. However, the long-term association of blood pressure (BP) and the risk of stroke-related mortality and the influence path of BP on stroke-related death remain unknown. The current study aimed to estimate the long-term causal associations between BP and stroke-related mortality and the potential mediating and moderated mediating model of the associations. METHODS: This is a 45-year follow-up cohort study and a total of 1696 subjects were enrolled in 1976 and 1081 participants died by the latest follow-up in 2020. COX proportional hazard model was used to explore the associations of stroke-related death with baseline systolic blood pressure (SBP)/diastolic blood pressure (DBP) categories and BP changes from 1976 to 1994. The mediating and moderated mediating effects were performed to detect the possible influencing path from BP to stroke-related deaths. E value was calculated in the sensitivity analysis. RESULTS: Among 1696 participants, the average age was 44.38 ± 6.10 years, and 1124 were men (66.3%). After a 45-year follow-up, a total of 201 (11.9%) stroke-related deaths occurred. After the adjustment, the COX proportional hazard model showed that among the participants with SBP ≥ 160 mmHg or DBP ≥ 100 mmHg in 1976, the risk of stroke-related death increased by 217.5% (hazard ratio [HR] = 3.175, 95% confidence interval [CI]: 2.297-4.388), and the adjusted HRs were higher in male participants. Among the participants with hypertension in 1976 and 1994, the risk of stroke-related death increased by 110.4% (HR = 2.104, 95% CI: 1.632-2.713), and the adjusted HRs of the BP changes were higher in male participants. Body mass index (BMI) significantly mediated the association of SBP and stroke-related deaths and this mediating effect was moderated by gender. CONCLUSIONS In a 45-year follow-up, high BP and persistent hypertension are associated with stroke-related death, and these associations were even more pronounced in male participants. The paths of association are mediated by BMI and moderated by gender.
Adult ; Blood Pressure/physiology* ; China/epidemiology* ; Follow-Up Studies ; Humans ; Hypertension ; Male ; Middle Aged ; Risk Factors ; Stroke