Objective To compare the efficacy of five Blastocystis staining method for staining cells with different morphologies and in different developmental stages to select appropriate staining techniques for specific scenarios and experimental purposes. Methods Blastocystis was stained using Wright-Giemsa, periodic acid-Schiff (PAS), Oil Red O, Nile Red, and Lugol's iodine solution respectively. The staining effects of the five method on Blastocystis were compared. Results Wright-Giemsa staining colored the cytoplasm and nucleus of Blastocystis bluish-purple, with other parts lightly stained; PAS staining rendered Blastocystis cells red; Oil Red O staining showed different structures within Blastocystis in varying intensities of orange-red; Nile Red enabled Blastocystis to fluoresce orange-yellow under a fluorescence microscope; Lugol's iodine solution stained the cytoplasm and nucleus a dark brownish-yellow. Conclusions Wright-Giemsa staining is recommended for clinical diagnosis. For metabolic studies, a combination of periodic acid-Schiff (PAS) staining and Oil Red O staining is suggested; PAS + Oil Red O staining can provide comprehensive morphological evidence for cell metabolism research. Nile Red staining is recommended for dynamic observation of live Blastocystis. Lugol's iodine staining is suitable for preliminary screening of large-scale samples.

Objective: To investigate the associations of anxiety, depressive symptoms and their comorbidity with neck-shoulder pain (NSP) and low back pain (LBP) among junior and senior high school students, thereby providing evidence for the promotion of physical and mental health. Methods: Stratified cluster random sampling method was used to select 638 junior and senior high school students in Taiyuan, Shanxi Province. The baseline survey (T1) was conducted in May 2025, and the follow up survey (T2) was conducted in October 2025. The Generalized Anxiety Disorder Scale-7, Patient Health Questionnaire-9, and Assessment of Spinal Health of Youths were used to assess anxiety/depressive symptoms and symptoms of NSP and LBP among junior and senior high school students. Multivariable modified Poisson regression models were applied to investigate the associations between anxiety, depressive symptoms and their comorbidity with NSP and LBP among junior and senior high school students. Results: The baseline survey showed that the detection rates of anxiety symptoms, depressive symptom, and their comorbidity symptoms among junior and senior high school students were 7.5%, 9.9% and 5.5%, respectively.The follow up results showed that the detection rates of NSP and LBP among junior and senior high school students were 10.7% and 10.8%. After adjusting for variables such as gender, school stage and boarding status, multivariable modified Poisson regression analyses indicated that depressive symptom at T1 was positively associated with NSP and LBP at T2 ( RR =2.41, 2.50; 95% CI =1.15-5.09,1.15-5.47,both P <0.05). The comorbidity of anxiety and depressive symptoms at T1 was positively associated with LBP at T2 ( RR =3.08, 95% CI =1.67-5.68, P <0.05). Conclusion Anxiety and depressive symptoms are positively associated with the occurrence of NSP and LBP among junior and senior high school students; among these, depressive symptoms and the comorbidity of anxiety and depressive symptoms show greater impacts.

Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Objective To explore the application of DRG mortality risk stratification method in evaluating perioperative healthcare quality,analyzes the distribution characteristics and influencing factors of perioperative mortality patients,and provide decision-making support for reducing perioperative mortality.Methods Collect perioperative case data from a tertiary hospital in Jiangxi Province from 2021 to 2023,and classify the risk of death for different DRG group.A chi-square test was used for univariate analysis,and a logistic regression model was used to analyze the factors affecting perioperative patient mortality.Results Among the death cases,the medium-and high-risk groups accounted for the highest proportion(59.31%),and the ratio of male to female was 2.2∶1.0,the most cases were over 65 years old(57.60%),and the top five Core DRG disease groups are neurological disorders,circulatory disorders,early grouping disorders,digestive disorders,and hepatobiliary and pancreatic disorders.Critical condition,serious complications or comorbidities,operation grade,relative weight,age,gender,and hospital day were independent influencing factors of perioperative death.All differences were statistically significant(P＜0.05).Conclusion To reduce perioperative mortality risks,it is necessary to enhance the treatment capacity for critically ill patients,optimize the dynamic management mechanism for surgical grading,and strengthen the risk management of emergency surgical medical care.

Objective:To explore the clinical efficacy and safety of endoscopic rubber band ligation (ERBL) and endoscopic injection sclerotherapy (EIS) in the treatment of grade I-III internal hemorrhoids, to provide a basis for the individualized treatment of patients with internal hemorrhoids in clinical practice.Methods:A retrospective cohort study was conducted. A total of 613 patients with grade I to III internal hemorrhoids who underwent ERBL or EIS at The Sixth Affiliated Hospital of Sun Yat-sen University from December 2019 to November 2023 were retrospectively collected. Inclusion criteria: (1) Patients diagnosed with symptomatic grade I-III internal hemorrhoids who failed conservative treatments such as diet adjustment and medication; (2) Patients who were unable or unwilling to receive surgical treatment due to multiple underlying systemic diseases. Exclusion criteria: (1) Patients with grade I-III internal hemorrhoids complicated with incarceration, thrombosis or other complications; (2) Patients with a history of hemorrhoid surgery; (3) Patients complicated with perianal abscess, anal fistula, active proctitis, rectal tumor, polyp, radiation proctitis or inflammatory bowel disease; (4) Patients with incomplete clinical data or lost to follow-up. This study was divided into the ERBL group and the EIS group based on different treatment. Baseline characteristics, postoperative effective rate, recurrence rate, pain score, anal distension, anal edema, complication rate, and treatment satisfaction were compared between the two groups.Results:After balancing with propensity score matching (PSM), a total of 313 patients were included, including 200 in the ERBL group and 113 in the EIS group. There were no statistically significant differences in baseline characteristics, such as gender, age, body mass index, Goligher classification, and laboratory test indicators, between the two groups (all P > 0.05), indicating that the two groups were comparable. The cure rates of the ERBL group and the EIS group were 64.0% (128/200) and 62.8% (71/113), respectively. The marked effective rates were 31.5% (63/200) and 34.5% (39/113), and the ineffective rates were 4.5% (9/200) and 2.7% (3/113), respectively. Statistical analysis revealed no statistically significant difference in efficacy between the two groups within 3 months after the operation (χ2=0.858, P=0.651). The recurrence rate of the ERBL group was lower than that of the EIS group [13.1% (25/191) vs. 18.2% (20/110)], but the difference was not statistically significant (χ2=1.424, P=0.233). Subgroup analysis showed that the recurrence rates of ERBL and EIS were similar in grade I-II internal hemorrhoids [10.3% (15/146) vs. 10.5% (9/86), χ2=0.002, P=0.963]; in grade III internal hemorrhoids, the recurrence rate of the ERBL group was lower than that of the EIS group [22.2% (10/45) vs. 45.8% (11/24), χ2=4.121, P=0.042]. Still, the difference was not statistically significant after Bonferroni correction (χ2=4.121, corrected P>0.025). Compared with the ERBL group, the EIS group had a lower pain score on the first day after operation [0 (0, 0) vs. 1 (0, 3), Z=-8.211, P<0.001] and a lower incidence of anal distension [25.7% (29/113) vs. 61.5% (123/200), χ2=37.122, P<0.001], with statistically significant differences. The total incidence of complications in the ERBL group was significantly higher than that in the EIS group [29.5% (59/200) vs. 4.4% (5/113), χ2=27.910, P<0.001], mainly reflected in postoperative bleeding [18.0% (36/200) vs. 3.5% (4/113), χ2=13.544, P<0.001] and urinary retention [8.5% (17/200) vs. 0, χ2=10.157, P=0.001], with statistically significant differences. There were no statistically significant differences in postoperative satisfaction, health status score, and the proportion of returning to everyday life within 1 month between the two groups (all P>0.05). Conclusions:Both ERBL and EIS are effective minimally invasive therapies for grade I-III internal hemorrhoids. EIS is preferred for grade I and II internal hemorrhoids to reduce complications, while ERBL shows a trend in reducing the recurrence rate of grade III internal hemorrhoids. Clinical decisions should be made comprehensively based on the classification, patient tolerance, and prognosis.

Objective:To explore the clinical efficacy and safety of endoscopic rubber band ligation (ERBL) and endoscopic injection sclerotherapy (EIS) in the treatment of grade I-III internal hemorrhoids, to provide a basis for the individualized treatment of patients with internal hemorrhoids in clinical practice.Methods:A retrospective cohort study was conducted. A total of 613 patients with grade I to III internal hemorrhoids who underwent ERBL or EIS at The Sixth Affiliated Hospital of Sun Yat-sen University from December 2019 to November 2023 were retrospectively collected. Inclusion criteria: (1) Patients diagnosed with symptomatic grade I-III internal hemorrhoids who failed conservative treatments such as diet adjustment and medication; (2) Patients who were unable or unwilling to receive surgical treatment due to multiple underlying systemic diseases. Exclusion criteria: (1) Patients with grade I-III internal hemorrhoids complicated with incarceration, thrombosis or other complications; (2) Patients with a history of hemorrhoid surgery; (3) Patients complicated with perianal abscess, anal fistula, active proctitis, rectal tumor, polyp, radiation proctitis or inflammatory bowel disease; (4) Patients with incomplete clinical data or lost to follow-up. This study was divided into the ERBL group and the EIS group based on different treatment. Baseline characteristics, postoperative effective rate, recurrence rate, pain score, anal distension, anal edema, complication rate, and treatment satisfaction were compared between the two groups.Results:After balancing with propensity score matching (PSM), a total of 313 patients were included, including 200 in the ERBL group and 113 in the EIS group. There were no statistically significant differences in baseline characteristics, such as gender, age, body mass index, Goligher classification, and laboratory test indicators, between the two groups (all P > 0.05), indicating that the two groups were comparable. The cure rates of the ERBL group and the EIS group were 64.0% (128/200) and 62.8% (71/113), respectively. The marked effective rates were 31.5% (63/200) and 34.5% (39/113), and the ineffective rates were 4.5% (9/200) and 2.7% (3/113), respectively. Statistical analysis revealed no statistically significant difference in efficacy between the two groups within 3 months after the operation (χ2=0.858, P=0.651). The recurrence rate of the ERBL group was lower than that of the EIS group [13.1% (25/191) vs. 18.2% (20/110)], but the difference was not statistically significant (χ2=1.424, P=0.233). Subgroup analysis showed that the recurrence rates of ERBL and EIS were similar in grade I-II internal hemorrhoids [10.3% (15/146) vs. 10.5% (9/86), χ2=0.002, P=0.963]; in grade III internal hemorrhoids, the recurrence rate of the ERBL group was lower than that of the EIS group [22.2% (10/45) vs. 45.8% (11/24), χ2=4.121, P=0.042]. Still, the difference was not statistically significant after Bonferroni correction (χ2=4.121, corrected P>0.025). Compared with the ERBL group, the EIS group had a lower pain score on the first day after operation [0 (0, 0) vs. 1 (0, 3), Z=-8.211, P<0.001] and a lower incidence of anal distension [25.7% (29/113) vs. 61.5% (123/200), χ2=37.122, P<0.001], with statistically significant differences. The total incidence of complications in the ERBL group was significantly higher than that in the EIS group [29.5% (59/200) vs. 4.4% (5/113), χ2=27.910, P<0.001], mainly reflected in postoperative bleeding [18.0% (36/200) vs. 3.5% (4/113), χ2=13.544, P<0.001] and urinary retention [8.5% (17/200) vs. 0, χ2=10.157, P=0.001], with statistically significant differences. There were no statistically significant differences in postoperative satisfaction, health status score, and the proportion of returning to everyday life within 1 month between the two groups (all P>0.05). Conclusions:Both ERBL and EIS are effective minimally invasive therapies for grade I-III internal hemorrhoids. EIS is preferred for grade I and II internal hemorrhoids to reduce complications, while ERBL shows a trend in reducing the recurrence rate of grade III internal hemorrhoids. Clinical decisions should be made comprehensively based on the classification, patient tolerance, and prognosis.

Objective To explore the application of DRG mortality risk stratification method in evaluating perioperative healthcare quality,analyzes the distribution characteristics and influencing factors of perioperative mortality patients,and provide decision-making support for reducing perioperative mortality.Methods Collect perioperative case data from a tertiary hospital in Jiangxi Province from 2021 to 2023,and classify the risk of death for different DRG group.A chi-square test was used for univariate analysis,and a logistic regression model was used to analyze the factors affecting perioperative patient mortality.Results Among the death cases,the medium-and high-risk groups accounted for the highest proportion(59.31%),and the ratio of male to female was 2.2∶1.0,the most cases were over 65 years old(57.60%),and the top five Core DRG disease groups are neurological disorders,circulatory disorders,early grouping disorders,digestive disorders,and hepatobiliary and pancreatic disorders.Critical condition,serious complications or comorbidities,operation grade,relative weight,age,gender,and hospital day were independent influencing factors of perioperative death.All differences were statistically significant(P＜0.05).Conclusion To reduce perioperative mortality risks,it is necessary to enhance the treatment capacity for critically ill patients,optimize the dynamic management mechanism for surgical grading,and strengthen the risk management of emergency surgical medical care.

This report described one patient of giant polycystic liver and polycystic kidney involving iliac fossa. Preoperative computed tomography (CT) revealed a large polycystic kidney occupying partially iliac fossa space. A decompression of lower pole of original kidney was planned for placing transplanted kidney. During total liver resection plus orthotopic liver transplantation, right polycystic kidney could move up on its own and iliac fossa space was released for placing transplanted kidney smoothly. Polycystic kidney shrunk markedly post-operation. It provided references for surgical planning of combined liver-kidney transplantation for this type of disease.

Objective:To delineate the surgical methodology and therapeutic paradigm of proximal tibial notch retrograde interlocking intramedullary nailing for ameliorating deformities due to osteofibrous dysplasia (OFD) in a pediatric population.Methods:A retrospective assessment was conducted on the medical records of individuals undergoing orthopedic osteotomy complemented by retrograde interlocking intramedullary nailing for OFD of the tibia from January 2016 to December 2019. The cohort comprised 15 patients, with a follow-up exceeding three years, documenting complete data sets. The patient profile included 8 males and 7 females, with 8 left-side and 7 right-side afflictions. The mean age at the time of surgery was 10.1±2.5 years, ranging from 7.1 to 12.6 years. Parameters measured were preoperative and postoperative imaging findings, which encompassed the scope of the lesion (longitudinal lesion length relative to tibial length), coronal and sagittal limb alignments, and lower limb length discrepancies.Results:The mean follow-up duration was 3.4±1.3 years, ranging from 3 to 6.6 years. Preoperatively, prominent anterior tibial arch deformities and limping were present, with 7 cases reporting fatigue-induced pain and 3 instances of pathological fractures. Post-surgery, pain symptoms were resolved, gait disturbances were improved in 9 patients, and completely resolved in 6. Tibial osteotomy or fracture healing of 15 patients averaged 3.9±0.7 months (range 3-5 months). The lesion range before surgery was 0.41±0.17, immediately after surgery was 0.38±0.17, and at the last follow-up was 0.30±0.16, with no statistical significance ( F=0.101, P=0.904). Lesion range showed no significant change throughout treatment, but radiographic density within the lesion notably increased post-surgery, suggesting bone improvement. The anterior tibial arch Angle was 28.30°±6.62° (range 20°-45°) before surgery, 4.73°±1.53° immediately after surgery, and 6.87°±1.36° at the last follow-up, with statistical significance ( F=159.739, P<0.001). A significant correction in the anterior tibial arch deformity was achieved and maintained postoperatively. There was no significant angular deformity of the tibia in the coronal plane before operation, and the medial proximal tibial angle (MPTA) and lateral distal tibial angle (LDTA) were 87.50°±1.46° and 88.30°±1.62°, 88.40°±1.46° and 88.70°±1.45° immediately after surgery, and 88.00°±1.39° and 89.10°±1.53° at the last follow-up, the differences were statistically significant ( F=1.741, P=0.188; F=1.016, P=0.371), there was no coronal deformity of tibia. The limb length discrepancy (LLD) was 0.60±0.98 cm before surgery, 0.18±0.93 cm at the last follow-up, with statistical significance ( t=0.096, P=0.761). There were no incidents of postoperative complications such as infection. Conclusion:In pediatric cases of tibial deformities attributed to osteofibrous dysplasia, a therapeutic strategy involving osteotomy for lower limb realignment, sans curettage or bone grafting of the lesion, followed by retrograde interlocking intramedullary nailing, yields favorable outcomes. Importantly, this implantation technique does not compromise the integrity of the proximal tibial epiphyseal plate in children and adolescents.

Objective:To explore the clinical effect of plating after lengthening to assist the consolidation of tibial shortening deformity in children.Methods:A retrospective analysis was conducted on 10 children with tibial shortening who were treated with circular external fixator lengthening and replacement with plate-assisted internal fixation (study group) in the Department of Pediatric Orthopedics of Tianjin Hospital from November 2019 to October 2022, and 16 children who were treated by circular external fixator only during the same period were enrolled as the external fixator group (control group). Among the 26 cases, there were 15 males and 11 females, 10 left knees and 16 right knees. The average age at the time of surgery was 8.8±4.7 years (range 5.1-13.8 years). The gender, side, age at the time of surgery were compared between the two groups. The extension length, external fixator index, healing index, Kolcaba comfort scale score, knee and ankle joint range of motion, complications and average total hospitalization costs were recorded.Results:There were no significant differences in gender, side, age at the time of surgery between the study group and control group. The extended lengths were 5.44±1.25 cm and 5.78±1.11 cm respectively without significant difference ( t=0.096, P=0.096). The external fixator index and healing index were 17.86±2.94 d/cm vs. 50.97±7.03 d/cm and 40.94±6.63 d/cm vs. 45.24±5.98 d/cm in study group and the control group with significant differences ( t=13.299, P<0.001; t=1.289, P=0.033). The Kolcaba comfort scale score of the children in the study group was higher than that of the control group, with significant difference ( t=6.821, P=0.001). Comparing the range of motion of the knee and ankle joints between the two groups, there were no significant differences before surgery, at the end of extension surgery, and at the final follow-up. When the extension end was healed, the range of motion of the knee joint was137.89°±4.40° vs. 114.09°±13.60° and ankle joint was 64.35°±5.50° vs. 56.65°±8.86° in the study group and control group with significant difference ( t=17.235, P<0.001; t=7.821, P=0.002). In the study group, 4 cases had pin tract infection, but no refracture occurred; in the control group, pin tract infection occurred in 13 cases, and refracture occurred in 2 cases after removal of the external fixator. The average total hospitalization cost of the study group was higher than that of the control group with significant difference ( t=3.745, P=0.036). Conclusion:The clinical effect of replacing plate-assisted internal fixator during the mineralization period of tibial shortening in children is reliable, and can significantly shorten the time for using external fixator. It is beneficial to the healing of the extended end of the osteotomy and the functional recovery of the knee and ankle joints. Being more comfortable for children with fewer complications, it is applicable for children who need long-distance extension, multiple extensions with limited joint function, poor tolerance for external fixators and low treatment compliance.