Objective:To characterize the longitudinal and dynamic high-density lipoprotein (HDL) trajectories in critically ill children and explore their correlation with clinical outcomes.Methods:Retrospective cohort study.All critically ill children admitted to the Pediatric Intensive Care Unit (PICU) of West China Hospital, Sichuan University from January 1, 2015 to October 1, 2020 were included in this retrospective study.Group-based trajectory modeling (GBTM) was applied to characterize the HDL trajectories in days 0-6 post-PICU admission and develop HDL trajectory groups.The in-hospital mortality rate was reported as frequency (%) and then compared by the Chi-square test or Fisher′s exact test between HDL trajectory groups.The length of stay (LOS) in the PICU was described by M( Q1, Q3), and its difference between HDL trajectory groups was evaluated by the Kruskal Wallis test.Logistic regression and multiple linear regression were used to determine the correlation between HDL trajectories and clinical outcomes.The primary outcome was in-hospital mortality rate, and the secondary outcome was LOS in the PICU. Results:A total of 4 384 critically ill children were ultimately enrolled in the study, and 6 HDL trajectory groups were developed based on GBTM analyses: group 1 (758 cases), the lowest HDL group; group 2 (1 413 cases), the low HDL group; group 3 (74 cases), the low-to-high HDL group; group 4 (621 cases), the medium HDL group; group 5 (1 371 cases), the high HDL group; and group 6 (147 cases), the highest HDL group.Logistic regression analysis showed that compared with critically ill children in group 1, those belonging to groups 2, 3, 4, 5, and 6 were at lower risks of in-hospital mortality with odds ratio ( OR): 0.475, 95%confidence interval ( CI): 0.352-0.641, P<0.001; OR: 0.093, 95% CI: 0.013-0.679, P=0.019; OR: 0.322, 95% CI: 0.208-0.479, P<0.001; OR: 0.263, 95% CI: 0.185-0.374, P<0.001, and OR: 0.142, 95% CI: 0.044-0.454, P=0.001, respectively.Multiple linear regression analysis revealed that compared with critically ill children in group 1, those belonging to groups 4, 5, and 6 had the trend of shorter LOS in PICU, and the β value and 95% CI were β: -4.332, 95% CI: -5.238- -3.426, P<0.001; β: -3.053, 95% CI: -3.809--2.297, P<0.001; β: -6.281, 95% CI: -7.842--4.721, P<0.001, respectively. Conclusions:The dynamic HDL trajectories during 0-6 days after PICU admission are associated with in-hospital mortality rate of critically ill children.The HDL trajectory at a persistently low level is associated with higher mortality, while the HDL trajectory at a persistently high level or with the trend from a low level rising to a high level shows a lower risk of mortality.It is suggested that the HDL trajectory model may become an indicator to predict the condition and prognosis of critically ill children.

Objective:To explore high-risk sexual behaviors of HIV/AIDS and related factors in young students in Guangzhou.Methods:A cross-sectional survey was conducted in 5 different types of Guangzhou colleges by convenience sampling with minimum number of classes per grade and 600 samples per school from September to November 2021. The R 4.2.2 software was used to consolidate databases. Simultaneously, a logistic regression model and a decision tree algorithm model, stratifying by whether sexual behaviors had occurred before, were constructed. In each layer, the prediction performance of the two models was evaluated through area under receiver operating characteristic and the confusion matrix, and then the model with high prediction performance was retained.Results:A total of 7 346 students were surveyed. The proportion of the respondents reporting sexual experience were 9.08% (667/7 346), in whom 26.24% (175/667) had risky sexual activity in the past year. The decision tree algorithm model performs well in predicting whether high-risk sexual behaviors have occurred in the past year. When the complexity parameter value is 0.018, and nsplit reaches 4, which means there are 5 leaf nodes in the model, the cross error of the tree will be the smallest. The first best grouping variable in the decision tree was whether to use condoms throughout the first sexual behavior. If condoms were used at their sexual debut, but homosexual practices have occurred in the past year, the probability of risky sexual behavior will increase. If homosexual practices have not occurred in the past year, but the age of sexual debut was below 18 years old while the period of HIV education was after high school, the probability of risk sexual behavior will also increase.Conclusions:AIDS-related risky behaviors of young students still deserved attention. The experience of sexual debut and whether AIDS-related health education has been received before the sexual debut were significant predictors for the occurrence of high-risk sexual behavior. The decision tree algorithm model has particular applicability for predicting and screening potential risk populations.

Objective:To analyze the impact of cecal ligation and puncture (CLP)-induced sepsis on the proliferation and differentiation of intestinal epithelial cells.Methods:① Animal experiment: sixteen male C57BL/6 mice were divided into sham operation group (Sham group) and CLP-induced sepsis model group (CLP group) by random number table method, with 8 mice in each group. After 5 days of operation, the jejunal tissues were taken for determination of leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5) and intestinal alkaline phosphatase (IAP) by polymerase chain reaction (PCR). The translation of LGR5 was detected by Western blotting. The expression of proliferating cell nuclear antigen (Ki67) was analyzed by immunohistochemistry. IAP level was detected by modified calcium cobalt staining and colorimetry. Immunofluorescence staining was used to detect the expression of Paneth cell marker molecule lysozyme 1 (LYZ1) and goblet cell marker molecule mucin 2 (MUC2). ② Cell experiment: IEC6 cells in logarithmic growth stage were divided into blank control group and lipopolysaccharide (LPS) group (LPS 5 μg/mL). Twenty-four hours after treatment, PCR and Western blotting were used to analyze the transcription and translation of LGR5. The proliferation of IEC6 cells were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining. The transcription and translation of IAP were detected by PCR and colorimetric method respectively.Results:① Animal experiment: the immunohistochemical results showed that the positive rate of Ki67 staining in the jejunal tissue of CLP group was lower than that of Sham group [(41.7±2.5)% vs. (48.7±1.4)%, P = 0.01]. PCR and Western blotting results showed that there were no statistical differences in the mRNA and protein expressions of LGR5 in the jejunal tissue between the CLP group and Sham group (Lgr5 mRNA: 0.7±0.1 vs. 1.0±0.2, P = 0.11; LGR5/β-actin: 0.83±0.17 vs. 0.68±0.19, P = 0.24). The mRNA (0.4±0.1 vs. 1.0±0.1, P < 0.01) and protein (U/g: 47.3±6.0 vs. 73.1±15.3, P < 0.01) levels of IAP in the jejunal tissue were lower in CLP group. Immunofluorescence saining analysis showed that the expressions of LYZ1 and MUC2 in the CLP group were lower than those in the Sham group. ②Cell experiment: PCR and Western blotting results showed that there was no significant difference in the expression of LGR5 between the LPS group and the blank control group (Lgr5 mRNA: 0.9±0.1 vs. 1.0±0.2, P = 0.33; LGR5/β-actin: 0.71±0.18 vs. 0.69±0.04, P = 0.81). The proliferation rate of IEC6 cells in the LPS group was lower than that in the blank control group, but there was no significant difference [positivity rate of EdU: (40.5±3.8)% vs. (46.5±3.6)%, P = 0.11]. The mRNA (0.5±0.1 vs. 1.0±0.2, P < 0.01) and protein (U/g: 15.0±4.0 vs. 41.2±10.4, P < 0.01) of IAP in the LPS group were lower than those in the blank control group. Conclusion:CLP-induced sepsis inhibits the proliferation and differentiation of intestinal epithelial cells, impairing the self-renewal ability of intestinal epithelium.

Objective:To investigate the risk perception for risky behavior of HIV/AIDS infection among young students and to analyze the related influencing factors.Methods:A cross-sectional survey was conducted in 5 different types of Guangzhou colleges from September to November 2021, in which convenience sampling and a minimum number of classes per grade and 600 samples per school were used according to the national unity program. Disordered multi-classification logistic regression was used to construct a risk perception model and analyze influencing factors in different risk perception levels.Results:A total of 7 346 young students were surveyed, and most rated themselves at low risk of HIV/AIDS infections (90.58%, 6 654/7 346). A total of 89.10% (6 545/7 346) of subjects' perception of their HIV/AIDS infection risk was consistent with their risk behavior, while 10.90% (801/7 346) was inconsistent. Among those inconsistent subjects, 19.10% (153/801) showed underestimating their risk , while 80.90% (648/801) seen overestimating their risk. Disordered multi-classification logistic regression analysis showed that, after controlling for other factors, compared with the non-sexual group, respondents whose first sex age under 18 had a higher rate of underestimating their risk of infection ( OR=129.39, 95% CI: 73.28-228.48), as well as a higher rate of overestimated their risk of infection ( OR=1.76, 95% CI: 1.04-2.99). First sexual intercourse at age 18 or older was a risk factor for underestimating risk ( OR=70.56, 95% CI: 42.72-116.53), but was not statistically associated with overestimating risk. Being female, other school type, non-heterosexual orientation, and self-rated HIV-related knowledge as fair or no knowledge were risk factors for overestimating risk but were not statistically associated with underestimating risk. Conclusions:Overall, young students in universities of Guangzhou have a good risk perception of HIV/AIDS infection. Individual factors, education factors and sexual experience will influence students' risk perception of HIV/AIDS infection. Raising the awareness rate of HIV/AIDS knowledge and delaying the age of first sexual intercourse will improve the risk perception ability of young students.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.

Objective:To evaluate the clinical outcomes of excimer laser atherectomy (ELA) in the treatment of diabetic foot with below-the-knee(BTK) lesions.Methods:The clinical data of 11 patients with diabetic foot with BTK lesions who underwent ELA at Department of Vascular Surgery,Zhongshan Hospital of Fudan University and Department of Vascular and Wound Treatment Center,Jinshan Hospital of Fudan University from September 2019 to May 2021 were retrospectively analyzed.There were 10 males and 1 female,aged 70.5 years(range:41 to 83 years).There were 20 lesions in 12 limbs,including 19 chronic total occlusion.All of the limbs were classified as Rutherford class 5 and suffered ulceration.The surgical efficacy,complications and ankle brachial index(ABI) after operation were record.Results:All patients underwent the operation successfully,the technical success rate was 12/12.No distal embolization,flow-limiting dissection,perforation or bailout stenting was occurred.The follow-up period was 8.2 months(range:3 to 13 months).The ABI increased from 0.58(range:0.24 to 1.57) before operation to 0.88(range:0.68 to 1.05) after operation.At 3 months after the operation,1 limb (1/12) underwent endovascular operation again due to restenosis,ulcers were healed in 5 limbs(5/12),and no amputation (limb/toe),death or loss of follow-up patients.Six months after the operation,2 patients were lost to follow-up and 2 died,ulcers were healed in 6 limbs(6/8),1 limb (1/8) underwent toe amputation due to prolonged healing of ulcers of toe.Conclusion:ELA is feasible and effective in the treatment of DF with BTK lesions,providing a new option of debulking atherectomy in such a group of patients.

Objective:To evaluate the clinical outcomes of excimer laser atherectomy (ELA) in the treatment of diabetic foot with below-the-knee(BTK) lesions.Methods:The clinical data of 11 patients with diabetic foot with BTK lesions who underwent ELA at Department of Vascular Surgery,Zhongshan Hospital of Fudan University and Department of Vascular and Wound Treatment Center,Jinshan Hospital of Fudan University from September 2019 to May 2021 were retrospectively analyzed.There were 10 males and 1 female,aged 70.5 years(range:41 to 83 years).There were 20 lesions in 12 limbs,including 19 chronic total occlusion.All of the limbs were classified as Rutherford class 5 and suffered ulceration.The surgical efficacy,complications and ankle brachial index(ABI) after operation were record.Results:All patients underwent the operation successfully,the technical success rate was 12/12.No distal embolization,flow-limiting dissection,perforation or bailout stenting was occurred.The follow-up period was 8.2 months(range:3 to 13 months).The ABI increased from 0.58(range:0.24 to 1.57) before operation to 0.88(range:0.68 to 1.05) after operation.At 3 months after the operation,1 limb (1/12) underwent endovascular operation again due to restenosis,ulcers were healed in 5 limbs(5/12),and no amputation (limb/toe),death or loss of follow-up patients.Six months after the operation,2 patients were lost to follow-up and 2 died,ulcers were healed in 6 limbs(6/8),1 limb (1/8) underwent toe amputation due to prolonged healing of ulcers of toe.Conclusion:ELA is feasible and effective in the treatment of DF with BTK lesions,providing a new option of debulking atherectomy in such a group of patients.

Through analysis of Lau Wan Yee's published papers,monographs,conference speeches,research designs / revisions for major hospitals in China,combined with analysis of articles revised by Academician Lau in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University for the past decade,authors came up with 9 points in Academician Lau's research ideas.Academician Lau has played a huge and leading role in improving clinical skills and scientific research levels,as well as in development and internationalization of hepatobiliary and pancreatic surgery in China.To summarize the research ideas of Lau is of great importance to the future generations of surgeons.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.