[Novel strategy of sepsis immunomodulation targeting dendritic cells].

Dendritic cells (DCs) are the major antigen-presenting cells that play critical roles in regulating both innate and acquire immune responses. In the state of sepsis, the number of DCs is obviously decreased with inhibited antigen presenting ability as well as abnormal cytokine secretion, thereby resulting in an impairment of T lymphocyte activation. Previous studies have demonstrated that the depletion and dysfunction of DCs appear to be the main causes associated with the development of sepsis-induced immunosuppression. Based on the characteristic changes of DCs in sepsis and analysis of recent research progress, the authors propose a novel strategy of immunomodulation targeting the apoptosis, differentiation, and dysfunction of DCs, in order to provide new ideas for the prevention and treatment of severe burns and trauma complicated with sepsis.

[Influence of family with sequence similarity 134, member B-mediated reticulophagy on lipopolysaccharide-induced apoptosis of mouse dendritic cells].

Objective: To investigate the influence of family with sequence similarity 134, member B (FAM134B)-mediated reticulophagy on lipopolysaccharide (LPS)-induced apoptosis of mouse dendritic cells (DCs), so as to provide a basis for improving the immune suppression of sepsis caused by wound infection and other factors. Methods: The experimental research methods were used. The DC line DC2.4 of the 3rd to 10th passage in the logarithmic growth stage was collected for experiments. DCs were divided into LPS stimulation 0 h (no stimulation) group, LPS stimulation 6 h group, LPS stimulation 12 h group, LPS stimulation 24 h group, and LPS stimulation 72 h group, which were cultured with 1 µg/mL LPS (the same concentration below) for the corresponding time. The protein expressions of FAM134B, microtubule-associated protein 1 light chain 3B (LC3B), and transporter protein SEC61B were determined by Western blotting, and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated (n=3). DCs were divided into phosphate buffer solution (PBS) group and LPS group for corresponding treatment. After 24 hours of culture, the expression of FAM134B and its co-localization with lysosomal probes and LC3B were detected using immunofluorescence method, while the number of autolysosomes in cells were observed through transmission electron microscope. DCs were divided into the FAM134B-knockdown group that were transfected with lentivirus containing small interfering RNA (siRNA) sequence of FAM134B gene and the empty vector group with empty lentivirus transfected. At post transfection hour 72, the fluorescence expression of cells was observed under the inverted fluorescence phase contrast microscope, meanwhile, the normally cultured DCs were set as blank control group, and the same observation was performed at the corresponding time point. DCs were divided into PBS alone group and LPS alone group, DCs successfully transfected with lentivirus containing siRNA sequence of FAM134B gene were divided into FAM134B-knockdown+PBS group and FAM134B-knockdown+LPS group, and DCs successfully transfected with empty lentivirus were divided into empty vector+PBS group and empty vector+LPS group. These cells were stimulated correspondingly and cultured for 24 hours. The protein expression of FAM134B was detected using Western blotting (n=3); the apoptotic rate of cells was determined by flow cytometry (n=3); the situation of apoptosis was observed by Hoechst staining, and the apoptotic rate was calculated (n=5); the protein expressions of cleaved cysteine aspartic acid specific protease-3 (caspase-3), B cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were detected using Western blotting, and the ratio of Bax/Bcl-2 was calculated (n=5). Data were statistically analyzed with one-way analysis of variance (ANOVA), least significant difference test, and ANOVA for factorial design. Results: Compared with those in LPS stimulation 0 h group, the protein expressions of FAM134B of cells in LPS stimulation 12 h group and LPS stimulation 24 h group were significantly increased (P<0.05), the protein expressions of SEC61B of cells in LPS stimulation 6 h group, LPS stimulation 12 h group, LPS stimulation 24 h group, and LPS stimulation 72 h group were significantly decreased (P<0.05), and the ratios of LC3B-Ⅱ/LC3B-Ⅰ of cells in LPS stimulation 24 h group and LPS stimulation 72 h group were obviously increased (P<0.05). As the most significant changes of three proteins were seen in the cells of LPS stimulation 24 h group, 24 h was used as the duration of subsequent LPS stimulation. After 24 hours of culture, the expression of FAM134B and its co-localization with LC3B and lysosomal probes in the cells of LPS group were all significantly enhanced, with a significant increase in the number of autolysosomes in comparison with those in PBS group. Both the empty vector group and the FAM134B-knockdown group showed high intensity fluorescence in the cells at post transfection hour 72, but the blank control group showed no fluorescence in the cells at the corresponding time point. After 24 hours of culture, the protein expression of FAM134B of cells in FAM134B-knockdown+PBS group was significantly lower than the expressions in PBS alone group and empty vector+PBS group (with P values all <0.05), the protein expression of FAM134B of cells in FAM134B-knockdown+LPS group was significantly lower than the expressions in LPS alone group and empty vector+LPS group (with P values all <0.05), the protein expression of FAM134B of cells in LPS alone group was significantly higher than that in PBS alone group (P<0.05), while the protein expression of FAM134B of cells in empty vector+LPS group was significantly higher than that in empty vector+PBS group (P<0.05). After 24 hours of culture, flow cytometry assay revealed that the apoptotic rate of cells in PBS alone group, LPS alone group, empty vector+PBS group, empty vector+LPS group, FAM134B-knockdown+PBS group, and FAM134B-knockdown+LPS group were (13.3±0.8)%, (32.6±4.3)%, (17.0±1.5)%, (51.7±3.3)%, (52.4±3.1)%, and (62.3±2.6)%, respectively. After 24 hours of culture, compared with those in LPS alone group and empty vector+LPS group, the protein expression of cleaved caspase-3, the ratio of Bax/Bcl-2, and the apoptotic rates of cells detected by flow cytometry and Hoechst staining were significantly increased in FAM134B-knockdown+LPS group (P<0.05); compared with those in the corresponding PBS treatment group, namely, PBS alone group, empty vector+PBS group, and FAM134B-knockdown+PBS group, the protein expression of cleaved caspase-3, the ratio of Bax/Bcl-2, and the apoptotic rates of cells detected by flow cytometry and Hoechst staining were significantly increased in LPS alone group, empty vector+LPS group, and FAM134B-knockdown+LPS group (P<0.05). Conclusions: The activation of reticulophagy mediated by FAM134B in mouse DCs is enhanced and peaked in 24 hours under LPS stimulation, and the activated reticulophagy has a significant inhibitory effect on cell apoptosis.

[Rehabilitation strategy for the improvement of long-term outcomes of patients after sepsis].

Survivors of sepsis still face high risks of secondary infection and mortality after hospital discharge. Meanwhile, the persistent cognitive, psychological, and physical disorders affect their long-term outcomes and life qualities. In the current review, we analyze the factors for the poor outcomes and discuss the beneficial rehabilitation strategies to improve the long-term outcomes of patients after sepsis, including psychological intervention, early mobility, nutrition support, and immune modulation, etc.

[Early predictive value of high density lipoprotein cholesterol for secondary acute kidney injury in sepsis patients].

Objective: To investigate the changes of high density lipoprotein cholesterol (HDL-C) in sepsis patients and its early predictive value for secondary acute kidney injury (AKI) in such patients. Methods: A retrospective case series study was conducted. From June 2019 to June 2021, 232 sepsis patients who met the inclusion criteria were admitted to the Second Hospital of Hebei Medical University, including 126 males and 106 females, aged 24 to 71 years. According to whether complicating secondary AKI, the patients were divided into non-AKI group (n=158) and AKI group (n=74). Data of patients between the two groups were compared and statistically analyzed with independent sample t test or chi-square test, including the sex, age, body mass index (BMI), body temperature, heart rate, primary infection site, combined underlying diseases, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and sepsis-related organ failure assessment (SOFA) score at admission, and the serum levels of C-reactive protein (CRP), procalcitonin, creatinine, cystatin C, and HDL-C measured at diagnosis of sepsis. The multivariate logistic regression analysis was performed on the indicators with statistically significant differences between the two groups to screen the independent risk factors for developing secondary AKI in 232 sepsis patients, and the joint prediction model was established based on the independent risk factors. The receiver operating characteristic (ROC) curve of the independent risk factors and the joint prediction model predicting secondary AKI in 232 sepsis patients were drawn, and the area under the curve (AUC), the optimal threshold, and the sensitivity and specificity under the optimal threshold were calculated. The quality of the above-mentioned AUC was compared by Delong test, and the sensitivity and specificity under the optimal threshold were compared using chi-square test. Results: The sex, age, BMI, body temperature, heart rate, primary infection site, combined underlying diseases, and CRP level of patients between the two groups were similar (P>0.05). The procalcitonin, creatinine, cystatin C, and scores of APACHE Ⅱ and SOFA of patients in AKI group were all significantly higher than those in non-AKI group (with t values of -3.21, -16.14, -12.75, -11.13, and -12.88 respectively, P<0.01), while the HDL-C level of patients in AKI group was significantly lower than that in non-AKI group (t=6.33, P<0.01). Multivariate logistic regression analysis showed that creatinine, cystatin C, and HDL-C were the independent risk factors for secondary AKI in 232 sepsis patients (with odds ratios of 2.45, 1.68, and 2.12, respectively, 95% confidence intervals of 1.38-15.35, 1.06-3.86, and 0.86-2.56, respectively, P<0.01). The AUCs of ROC curves of creatinine, cystatin C, HDL-C, and the joint prediction model for predicting secondary AKI in 232 sepsis patients were 0.69, 0.79, 0.89, and 0.93, respectively (with 95% confidence intervals of 0.61-0.76, 0.72-0.85, 0.84-0.92, and 0.89-0.96, respectively, P values all below 0.01); the optimal threshold were 389.53 µmol/L, 1.56 mg/L, 0.63 mmol/L, and 0.48, respectively; the sensitivity under the optimal threshold were 76.6%, 81.4%, 89.7%, and 95.5%, respectively; the specificity under the optimal threshold values were 78.6%, 86.7%, 88.6%, and 96.6%, respectively. The AUC quality of cystatin C was significantly better than that of creatinine (z=2.34, P<0.05), the AUC quality and sensitivity and specificity under the optimal threshold of HDL-C were all significantly better than those of cystatin C (z=3.33, with χ2 values of 6.43 and 7.87, respectively, P<0.01) and creatinine (z=5.34, with χ2 values of 6.32 and 6.41, respectively, P<0.01); the AUC quality and sensitivity and specificity under the optimal threshold of the joint prediction model were all significantly better than those of creatinine, cystatin C, and HDL-C (with z values of 6.18, 4.50, and 2.06, respectively, χ2 values of 5.31, 7.23, 3.99, 6.56, 7.34, and 4.00, respectively, P<0.05 or P<0.01). Conclusions: HDL-C level in sepsis patients with secondary AKI is significantly lower than that in patients without secondary AKI. This is an independent risk factor for secondary AKI in sepsis patients with a diagnostic value being superior to that of creatinine and cystatin C. The combination of the aforementioned three indicators would have higher predicative valuable for secondary AKI in sepsis patients.

[Analysis on prediction power of HIV infection risk assessment tool in men who have sex with men in Guizhou province].

Objective: To evaluate the prediction power of HIV infection risk assessment tool and the applicability in MSM in Guizhou province. Methods: MSM were recruited through snowball sampling method. Questionnaire surveys were conducted among the MSM using HIV infection risk assessment tool, and combined with HIV serologic test results, the risk prediction power of HIV infection risk assessment tool was evaluated. Results: A total of 3 379 MSM were recruited from January 2018 to December 2019 in Guizhou. The HIV infection rate was 3.3%(111/3 379). The mean risk scores of HIV positive and HIV negative MSM were (12.15±3.08) and (12.07±3.07), respectively. The difference in risk score was significant between MSM with different HIV status (t=8.69, P<0.001). According to the principle of decision tree, individual risk scores were divided into following three categories: ≤11.96, 11.97-14.80 and >14.80, the HIV infection rate was 0.8%, 4.3% and 8.6% respectively, suggesting that the higher the individual risk score was, the higher the HIV infection rate was (trend χ2=88.18, P<0.001). Multivariate logistic regression analysis showed that the higher the individual risk score was, the higher the risk of HIV infection was. Compared to the total score ≤11.96, the aOR values at total scores of 11.97-14.80 and >14.80 were 6.34 (95%CI: 3.38-11.88) and 14.07(95%CI: 7.44-26.61), respectively. The risk of HIV infection in Miao ethnic group was higher than that in Han ethnic group (aOR=1.83, 95%CI:1.04-3.21), and the risk of HIV infection in those with education level of primary school and below was higher than that in undergraduates or those with education level of junior college and above (aOR=2.50, 95%CI:1.06-5.88), and the risk of HIV infection was higher in those who had bisexual behaviors than in those who had homosexual behaviors (aOR=1.95, 95%CI:1.19-3.19). The risk of HIV infection was higher in those who had never received HIV testing (aOR=1.53, 95%CI:1.01-2.33). The area under the receiver operating characteristic (ROC) curve and area under ROC (AUC) for HIV infection prediction was 0.751 (95%CI:0.710-0.792, P<0.001). The maximum Youden's index was individual risk score of 12.56, and the sensitivity of the risk assessment tool was 0.838, and its specificity was 0.412. Conclusions: The results of HIV infection risk assessment tool in Guizhou indicated that in MSM the higher the individual risk score, the higher the risk of HIV infection is. The tool can be used to evaluate the risk of HIV infection in MSM, but the specificity should be improved.

[New understanding on the immunity for severe infections and complications in burns and trauma].

The mechanisms of burn/trauma related infection are complicated, involving uncontrolled inflammation, immune disorder, coagulation abnormality, and pathophysiological changes of multiple systems or organs. Although active management of primary diseases, application of antibiotics, and organ support treatment are the basis for the management of burn/trauma related infection, immune modulation opens up a new direction for the intervention of burn/trauma related infection and the relevant complications. In-depth understanding of inflammation-immune response and its regulatory mechanisms, and exploring new early warning biomarkers and immune interventional strategies are of great significance for improving the level of treatment of clinical burn/trauma related infections and improving the prognosis of patients.

[Deepening the understanding of the diagnostic system and therapeutic strategy for burn sepsis].

Burn patients with large area of skin defect are prone to cause wound infection, severe fluid loss, and hypermetabolism, etc, which lead to sequential dysfunction of multiple systems and easily induce severe systemic infections and sepsis. At present, sepsis has been the leading causes of death in severe burn patients, and its early diagnosis and timely treatment are critical for successful treatment of patients. As burn sepsis has unique pathophysiological characteristics, the diagnostic criteria for general sepsis lack specificity for burn sepsis. Therefore, to understand the pathogenesis of burn will help deepen the understanding of the diagnostic system and interventional way of burn sepsis, thus developing more precise and intelligent therapeutic strategy.

[A survey on sexual needs and factors of HIV risky sexual behaviors among elderly men at different ages in two communities of Qiandongnan Miao and Dong autonomous prefecture].

Objective: To understand sexual needs and factors of risky sexual behaviors among elderly men at different ages in two communities of Qiandongnan Miao and Dong autonomous prefecture and provide basis for targeted HIV prevention and intervention. Methods: Two communities in the prefecture were selected as study sites. Questionnaire surveys were carried out among elderly men aged 50 and over who visited or consulted in the communities from June to December 2018, and they were tested for HIV and syphilis antibodies. Results: Among 400 elderly men, 209 (52.2%) were 50-64 years old, and 191(47.8%) were above 65 years old. They were mainly Miao people, accounting for 66.3% (265/400), and 235 (58.8%) had an education no more than 6 years. HIV awareness of the two age groups were only 25.8% (54/199) and 26.2% (50/191), respectively. Among those aged 50-64, 142 (68.0%) felt normal sexual desire, and 153 (73.6%) reported penile erections or erections in most cases whenever sex, and 52.9% (110) ejaculated most of the time. HIV prevalence was 1.0% (4/400). Compared with the over 65-year-old group, the proportion of having sex with spouse/stable partners (89.5%, 179/200), proportion of no condom use with their spouse/stable sexual partners during the most recent sex (93.8%, 168/179), proportion of having casual sex (11.0%, 23/209) and commercial sex (3.8%, 8/209) were all higher among 50-64 age group. In comparison to those aged over 65 years old, average monthly income>3 000, and use of sex helper, aged 50-64 (OR=2.70, 95%CI: 1.22-5.95), average monthly income ≤1 000 yuan (OR=2.79, 95%CI: 1.25-6.21), and no use of sex helper (OR=3.78) (95%CI: 1.65-8.67) were related factors of HIV risky sexual behavior last time. Conclusion: Elderly men in the minority prefecture had low HIV awareness. Compared with those≥65 years old, the 50-64 age group had more active sexual behaviors and higher sexual needs. Those from 50-64 age group, with lower economic level and good sexual ability were more likely to have HIV risky sexual behaviors.

[Influence of Xuebijing injection and its component paeoniflorin on immune function and survival rate of septic rats].

Objective: To explore the influence of Xuebijing injection (hereinafter referred to as Xuebijing) and its component paeoniflorin on immune function of regulatory T cells (Tregs) of spleen and survival rate of septic rats. Methods: (1) CD4(+) CD25(+) Tregs and CD4(+) T cells were isolated and purified from spleens of three 9 to 12 weeks old Sprague-Dawley male rats (the same age, breed, and gender below) by immunomagnetic beads. According to the random number table (the same grouping method below), CD4(+) CD25(+) Tregs were divided into blank control group, simple CD3/CD28 group, simple endotoxin/lipopolysaccharide (LPS) group, LPS+ Xuebijing group, and LPS+ paeoniflorin group, with 6 wells in each group. The cells in simple CD3/CD28 group, simple LPS group, LPS+ Xuebijing group, and LPS+ paeoniflorin group were cultured in RPMI 1640 medium containing fetal bovine serum in volume fraction of 10%, 1.25 µg CD3, and 2.5 µg CD28 for 24 hours. Then 1 µg/mL LPS in the volume of 1 µL was added to the cells in simple LPS group, LPS+ Xuebijing group, and LPS+ paeoniflorin group. Moreover, 5 mg/mL Xuebijing in the volume of 1 µL and 80 µmol/L paeoniflorin in the volume of 1 µL were added to the cells in LPS+ Xuebijing group and LPS+ paeoniflorin group, respectively, which were cultured for another 72 hours. Cells in blank control group were routinely cultured in RPMI 1640 medium containing fetal bovine serum in volume fraction of 10% for 96 hours. The expressions of cytotoxic T lymphocyte antigen 4 (CTLA-4) and forkhead wing-link transcription factor 3 (Foxp3) and apoptosis of CD4(+) CD25(+) Tregs were measured by flow cytometry. The interleukin-10 (IL-10) level from culture supernatant of CD4(+) CD25(+) Tregs was determined by enzyme-linked immunosorbent assay (ELISA). CD4(+) T cells were divided into blank control' group, simple CD3/CD28' group, simple LPS' group, LPS+ Xuebijing' group, and LPS+ paeoniflorin' group, with 6 wells in each group. After being cocultured with the corresponding CD4(+) CD25(+) Tregs treated as before for 72 hours, the proliferative activity of CD4(+) T cells was measured by flow cytometry, and IL-4 level from culture supernatant of CD4(+) T cells was determined by ELISA. (2) One hundred and twenty rats were divided into sham surgery group, simple sepsis group, sepsis+ Xuebijing group, and sepsis+ paeoniflorin group, with 30 rats in each group. The septic rat model was reproduced by cecal ligation and puncture surgery in simple sepsis group, sepsis+ Xuebijing group, and sepsis+ paeoniflorin group. In sham surgery group, the rats were only performed with laparotomy to simulate surgery. In sepsis+ Xuebijing group, the rats were given post-surgical injection of 4 mL/kg Xuebijing through tail vein, twice a day. In sepsis+ paeoniflorin group, the rats received 978 µg paeoniflorin via tail vein, twice a day. The survival rates of rats in the four groups on post surgery day 1, 2, 3, 4, 5, 6, and 7 were observed and recorded. The surviving cure of Kaplan-Meier was drawn. Data were statistically analyzed with one-way analysis of variance, least significant difference t test. The surviving curve was analyzed by Log-rank (Mantel-Cox) test. Results: (1) Compared with those in blank control group, the expressions of CTLA-4 and Foxp3 of CD4(+) CD25(+) Tregs (t=27.19, 17.00, P<0.01) and IL-10 level from culture supernatant (t=40.76, P<0.01) were significantly increased in rats in simple LPS group. Compared with those in simple LPS group, the expressions of CTLA-4 and Foxp3 of CD4(+) CD25(+) Tregs (t(LPS+ Xuebijing group)=31.03, 11.27, t(LPS+ paeoniflorin group)=5.79, 5.64, P<0.01) and IL-10 level from culture supernatant (t=15.49, 4.20, P<0.01) was significantly decreased in LPS+ Xuebijing group and LPS+ paeoniflorin group. Compared with that in blank control group, the apoptosis rate of CD4(+) CD25(+) Tregs in simple LPS group was significantly declined (t=6.02, P<0.01). Compared with the rate in simple LPS group, the apoptosis rates of CD4(+) CD25(+) Tregs in LPS+ Xuebijing group and LPS+ paeoniflorin group were significantly increased (t=20.32, 8.60, P<0.01). (2) Compared with those in simple CD3/CD28' group, the proliferative rate of CD4(+) T cells was significantly decreased in simple LPS' group (t=22.47, P<0.01), while IL-4 level from culture supernatant was significantly elevated (t=3.51, P<0.01). Compared with those in simple LPS' group, the proliferative rates of CD4(+) T cells in LPS+ Xuebijing' group and LPS+ paeoniflorin' group were significantly increased (t=16.31, 11.48, P<0.01), while IL-4 level from culture supernatant showed no obvious change. (3) The post-operative 7-day survival rates of rats in sham surgery group, simple sepsis group, sepsis+ Xuebijing group, sepsis+ paeoniflorin group were 100% (30/30), 30% (9/30), 57% (17/30), and 47% (14/30), respectively. Compared with that in simple sepsis group, the survival rate within post-operative 7-day of rats in sepsis+ Xuebijing group was significantly higher (χ(2)=4.34, P<0.05), while the survival rate within post-operative 7-day of rats in sepsis+ paeoniflorin group showed no obvious change. Conclusions: Both Xuebijing and its component paeoniflorin are capable of reversing sepsis-induced inhibitory immune function and apoptotic resistant of Tregs in rats, and further improving the proliferative activity of T cells. In addition, the effect of paeoniflorin on improvement of survival rate of rats with sepsis is weaker than Xuebijing.

[Survival time and related factors on HIV/AIDS patients in Guizhou province from 1995 to 2018].

Objective: To examine the survival time and related factors on HIV/AIDS patients in Guizhou province from 1995 to 2018. Methods: A retrospective cohort study was conducted to analyze the HIV/AIDS case from 1995 to 2018 in Guizhou province with data gathered from the "Chinese National Comprehensive HIV/AIDS Prevention and care Information system". Survival rate was calculated by life table and survival time was estimated by Kaplan-Meier. Related factors on survival time were analyzed by Cox regression model. Results: A total of 53 232 HIV/AIDS cases were included in the study, with the mortality rate as 8.53/100 person-years (14 210/166 679.18), median survival time as 10.20 (95%CI: 9.91-10.48) years, and survival rates of 1, 5, 10 and 20 years as 0.85, 0.68, 0.51, 0.36, 0.19 respectively. Results from the multivariate Cox regression analysis showed that factors as: being male (compared with females, aHR=0.757, 95%CI: 0.727-0.788), with antiviral treatment (ART) (compared with those without ART, aHR=0.173, 95%CI: 0.165-0.181), CD(4)<200 cells/µl[compared with CD(4)(+)T cells (CD(4)) ≥200 cells/µl, aHR=0.410, 95%CI: 0.387-0.435], age ≥45 (compared with age<45, aHR=1.506, 95%CI: 1.193-1.901), illiterate (compared with having high school education or above, aHR=0.904, 95%CI: 0.832-0.982), unmarried (compared with divorced or widowed, aHR=0.896, 95%CI: 0.848-0.946), through heterosexual transmission (compared with homosexual transmission, aHR=0.555, 95%CI: 0.487-0.632), ethnic minorities (compared with Hans, aHR=1.185, 95%CI: 1.114-1.262), and farmers/migrant workers (compared with domestic/unemployed,aHR=0.874, 95%CI: 0.834-0.916,) etc., were related to the survival time of HIV/AIDS, in Guizhou province. Conclusions: The mortality rate of HIV/AIDS in Guizhou province appeared relatively high, but with no obvious downward trend seen in the last years. Factors as being male, age ≥45, low education level, ethnic minorities, CD(4)<200 cells/µl were identified as related to the HIV/AIDS survival time. We would suggest that treatment and follow-up management programs should be strengthened to improve the quality of life among these patients.

[Update in immune regulatory dysfunction of dendritic cells in sepsis].

Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. Further development of sepsis usually leads to septic shock or even death. Many previous studies have focused on the abnormal reactions of monocytes/macrophages, neutrophils, complement system, or cytokine inflammation in sepsis. Many evidences in recent years suggest that dendritic cells, as the most powerful antigen-presenting cells in innate immune system of body, play important role during the process of immune disorders of sepsis. In this article, I review the main classification, immune function, monitoring method, regulatory pathways of dendritic cells and their clinical significance in immune disorders of sepsis, so as to find new strategies for immune regulation of sepsis.

[Advances in the research of the relationship between skin regulatory T cells and wound healing and immune diseases].

As the body's largest organ, skin harbors a large amount of immune cells to regulate both innate and adaptive immune responses. Regulatory T cells (Tregs), as a subset of T lymphocytes with negative regulatory functions, play an important role in maintaining the immune homeostasis of different tissue. However, researches of skin Tregs are largely limited and uncompleted as compared with other tissue. In recent years, a comprehensive understanding is increasingly showing the specialized functions of Tregs in skin, including the orchestration of tissue wound healing, involvement in hair follicle recycling, and modulation of proper immune homeostasis. In this review, we outline the classification and characteristics of Tregs in skin, distribution, migration routes, immune effects, and relationship with wound healing, which aims to deepening our understanding towards the immunological effects of T lymphocytes subsets in skin and its regulatory pathways.

[Advances in the research of effects and regulatory mechanism of regulatory T cells in tissue injury and repair].

The repair strategy after organs injuries has always been a hot topic in the field of regenerative medicine. Traditional injury repair measures mainly promote tissue repair through mesenchymal stem cells and various growth factors, but these strategies have been constrained in the aspects of security and economy. Hence, there is an urgent need to find new ways to promote tissue repair and regeneration. There have been a lot of evidences showing that the immune system plays an important role in tissue regeneration and repair. In recent years, more and more studies have been done on adaptive immunity in tissue repair, especially the regulatory T cells. Some evidences indicate that regulatory T cells participate in damage tissue repair and regeneration of multiple organs and tissue. This review briefly introduces the new advances in the repair effects and regulatory mechanism of regulatory T cells in different organ injuries, in order to provide new ideas for designing advanced repair materials with good immunoregulatory functions.

[Mortality and influencing factors on injecting drug users with HIV/AIDS in Guizhou province, 1996-2015].

Objective: To understand the mortality and influencing factors on injecting drug users (IDUs) with HIV/AIDS, in Guizhou province, 1996-2015. Methods: A retrospective cohort study was conducted on IDUs with HIV/AIDS that were reported through national comprehensive HIV/AIDS information system, in Guizhou province during 1996-2015. Cox proportional hazard regression model was used to analyze the influencing factors on the mortality of HIV/AIDS. Results: A total of 3 958 cases of IDUs with HIV/AIDS were recruited in this study, with all-cause mortality rate of 44.01% (1 742/3 958) and total mortality rate of 7.80/100 person-years, respectively. The median survival time between diagnosis and death was 8.08 years. Mortality rate was 3.57/100 person-years in the group receiving antiretroviral therapy (ART). The mortality appeared to be 4.08/100 person-years in the group who were on methadone maintenance treatment (MMT). Data from the multiple regression analysis indicated that factors of gender, ethnicity, age when HIV/AIDS diagnosis was made, CD(4)(+)T lymphocyte (CD(4)) count at the first testing, ART and MMT were significantly associated with deaths among these people. The risk of death in females was 0.82 times (95%CI: 0.69-0.98) higher than that in males. The risk of deaths among the ethnic minority subjects was 1.39 times (95%CI: 1.21-1.60) higher than that of the Hans. The risk of death appeared to be 2.44 times higher (95%CI: 1.07-5.56) in the over-50-year of age group than in the <20 year-old group, when HIV/AIDS was diagnosed for the first time. The risk of death in CD(4) ≥500/µl group in the first time was 0.27 times (95%CI: 0.22-0.32) more than CD(4) <200/µl group in the firs time. The risk of death in cases who were treated with ART or MMT was 2.83 times (95%CI: 2.45-3.26) and 1.35 times (95%CI: 1.15-1.59) higher than those who did not receive any treatment, respectively. Conclusion: Higher risks on death seemed to be related to the following factors: being male, older age at the time of diagnosis, lower CD(4) at diagnosis, not on ART or MMT among the IDUs with HIV/AIDS in Guizhou province, between 1996-2015.

[Precision evaluation of immune status and its significance in sepsis after burns or trauma].

Sepsis induced by major burns, trauma, and hemorrhage, remains a major cause of death of patients in intensive care units, and it may result in both the widespread activation and dysfunction of the innate as well as adaptive responses in host immune system. A large amount of information concerning subsets of innate and adaptive immune cells in sepsis has implicated that these cells, including neutrophils, macrophages, dendritic cells, T lymphocytes, regulatory T lymphocytes, and natural killer cells, have profound effects on immunoreactivity during acute insults or sepsis through modulating multiple receptor expressions or cytokine secretion, in turn contributing to the development and outcome of sepsis. It is of great significance that precision monitoring of immune function and the related indicators might help to assess the risk of secondary infection, the prognosis of septic patients, and guide the treatment of septic complications.

[Influence of vagus nerve on multiple organ function and immune reaction of T lymphocytes in septic rats].

Objective: To explore influence of vagus nerve on multiple organ function and immune reaction of T lymphocytes in septic rats. Methods: Forty Sprague-Dawley rats were divided into sham injury group, sepsis group, vagus nerve stimulation (VNS) group, and vagotomy (VGX) group, according to the random number table, with 10 rats in each group. Rats in sepsis group, VNS group, and VGX group were inflicted with sepsis by cecal ligation and puncture (CLP). Rats in VNS group were given electrical stimulation on left cervical vagus nerve for 15 min right after CLP. Rats in VGX group underwent vagotomy of left cervical vagus nerve at 30 min before CLP. At 24 h after CLP, serum of rats was collected to detect levels of alanine transaminase (ALT), aspartate aminotransferase (AST), glycocholic acid (CG), creatine kinase (CK), myocardial creatine kinase (CK-MB), blood urea nitrogen (BUN), and serum creatinine by fully automatic biochemistry analyzer. The left lung of rats was collected to determine wet or dry mass, and wet to dry (W/D) ratio was calculated. The right lung of rats was collected to measure the activity of pulmonary myeloperoxidase (MPO) by enzyme linked immunosorbent assay (ELISA). Spleen of rats was collected to determine the proliferative activity of CD4(+) T lymphocytes by cell counting kit 8, and real-time fluorescence quantitative polymerase chain reaction and ELISA were used to quantify mRNA expressions and levels of interleukin 2 (IL-2), interferon-γ, and IL-4, respectively. Data were processed with one-way analysis of variance and Tukey's honest significant difference test. Results: (1) The levels of serum ALT, AST, CG, CK, CK-MB, BUN, and creatinine, pulmonary W/D ratio, as well as MPO activity of rats in sepsis group were significantly higher than those in sham injury group and VNS group (P<0.01) and were significantly lower than those in VGX group (P<0.01). (2) The proliferative activity of CD4(+) T lymphocytes of rats in sepsis group was 0.93±0.03, which was significantly lower than 1.54±0.07 of rats in sham injury group (P<0.01). The proliferative activity of CD4(+) T lymphocytes of rats in VNS group was 1.15±0.15, which was significantly higher than that of rats in sepsis group (P<0.01). The proliferative activity of CD4(+) T lymphocytes of rats in VGX group was 0.75±0.06, which was obviously lower than that of rats in sepsis group (P<0.01). (3) In comparison with those of rats in sham injury group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in sepsis group were markedly decreased (P<0.01), while the level of IL-4 was significantly increased (P<0.01). In comparison with those of rats in sepsis group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in VNS group were obviously increased (P<0.01), while the level of IL-4 was markedly decreased (P<0.01). As compared with those of rats in sepsis group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in VGX group were markedly decreased (P<0.01), while the level of IL-4 was significantly increased (P<0.01). (4) As compared with those of rats in sham injury group, expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in sepsis group were markedly decreased (P<0.01), while expression of IL-4 mRNA was significantly increased (P<0.01). Expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in VNS group were obviously increased when compared with those of rats in sepsis group (P<0.01), while expression of IL-4 mRNA was markedly decreased (P<0.01). In comparison with those of rats in sepsis group, expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in VGX group were markedly decreased (P<0.01), while expression of IL-4 mRNA was significantly increased (P<0.01). Conclusions: Electrical stimulation of vagus nerve can significantly improve multiple organ dysfunction and reverse immunosuppression of T lymphocytes in septic rats, while vagotomy of vagus nerve may enhance the susceptibility of rats to sepsis.

[A clinical study of serum protein markers in patients with 1-bromopropane poisoning].

Objective: To investigate the changes in protein expression in patients with 1-bromopropane (1-BP) poisoning using high-throughput proteomic technique and to screen out protein markers. Methods: Serum samples were collected from 3 patients with 1-BP poisoning and 15 controls. The label-free proteomic tech-nique was used for the quantitation and identification of proteins expressed in these samples, and the results were compared between the patients with 1-BP poisoning and the control population. The bioinformatics tools were used to analyze the function of differentially expressed proteins. Results: Compared with the control popula-tion, the patients with 1-BP poisoning had >2-fold upregulation of 38 proteins and >2-fold downregulation of 68 proteins. The differentially expressed proteins were mainly involved in immune response, signal transduction, and stress response. Conclusion: The proteins screened out may be potential protein markers for 1-BP poison-ing, which provides reliable and precise methods and thoughts for the diagnosis of 1-BP poisoning.

[Effects of rabbit adipose-derived mesenchymal stem cells on the healing of skin deep partial-thickness scald wound of rabbit].

OBJECTIVE: To investigate the effects of local injection of rabbit adipose-derived mesenchymal stem cells (ADSCs) on the healing of skin deep partial-thickness scald wound of rabbit. METHODS: ADSCs were isolated from adipose tissue of one New Zealand rabbit and then sub-cultured. ADSCs of the third passage were used in the following experiments. Twenty-four rabbits were divided into ADSCs group (n=12) and control group (n=12) according to the random number table, and one deep partial-thickness scald wound with diameter of 5 cm on the two sides of the back near the buttocks was made. From post injury day (PID) 2, 2 mL suspension of EdU-labeled ADSCs with the number of 5×10(5) per mL was subcutaneously injected in wounds of rabbits in ADSCs group, while the rabbits in control group were given 2 mL serum-free DMEM until the wounds were healed. Wound healing processes of rabbits in two groups were observed every day, and the healing time was recorded. On PID 7, 14, 21, and 28, areas of wound of three rabbits in two groups were measured and the healing rates were calculated, respectively. The healed wound tissue was harvested to observe the morphology by HE staining, and the expression of collagen fiber was observed by Masson staining. The distribution of EdU-labeled ADSCs in healed wound tissue on PID 28 was observed by inverted fluorescence microscope. The expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) of healed wound tissue on PID 7, 14, and 21 were detected by enzyme-linked immunosorbent assay. Data were processed with analysis of variance of factorial design and paired samples t test. RESULTS: (1) The wound healing time of rabbits in ADSCs group was (19.5±1.1) d post injury, which was significantly shorter than that in control group [(23.3±1.5) d, t=4.50, P<0.05]. On PID 7, wounds of rabbits in two groups were dry with no obvious exudation, and redness and swelling around wounds disappeared gradually, the wound healing rate of rabbits in ADSCs group was (15.1±2.4)%, which was close to that in control group [(13.7±3.1)%, t=1.20, P>0.05]. On PID 14, wounds of rabbits in ADSCs group were dry and scabbed obviously, and the wound healing rate was (73.1±5.7)%, while wounds of rabbits in control group were little scabbed with little exudation, and the wound healing rate was significantly lower than that in ADSCs group [(52.9±5.1)%, t=8.06, P<0.01]. On PID 21, wounds of rabbits in ADSCs group were generally healed, and the wound healing rate was (95.6±3.0)%, while a few wounds still existed in rabbits of control group, and the wound healing rate was significantly lower than that in ADSCs group [(78.6±3.7)%, t=9.73, P<0.01]. On PID 28, wounds of rabbits in two groups were totally healed with the healing rate of 100%, and texture and microvascular responses of healed wound tissue in ADSCs group were better than those in control group. (2) On PID 7, fibroblasts in healed wound tissue of rabbits in two groups were all increased, and there were little vascular and collagen fiber proliferation with no obvious differences. On PID 14, the number of fibroblasts in healed wound tissue of rabbits in ADSCs group was more than that in control group, and the collagen fibers in healed wound tissue of rabbits in ADSCs group were arranged in dense and uniform, while those in control group were sparse and irregular. On PID 21, skin layers were differentiated in healed wound tissue of rabbits in two groups, and collagen fibers in healed wound tissue of rabbits in ADSCs group were still denser than that in control group. On PID 28, newborn skin was well differentiated in healed wound tissue of rabbits in ADSCs group, which was better than that in control group. There were a lot of thick collagen fibers in healed wound tissue of rabbits in two groups, and EdU-labeled ADSCs were involved in skin texture of rabbits in ADSCs group. (3) The expressions of VEGF and EGF in healed wound tissue of rabbits in two groups were similar on PID 7 (with t values respectively 0.70 and 0.91, P values above 0.05), which in ADSCs group were significantly higher than those in control group on PID 14 and 21 (with t values from 2.85 to 4.81, P values below 0.01). CONCLUSIONS: The transplantation of ADSCs can promote the wound healing of skin deep partial-thickness scald wound of rabbit and shorten the wound healing time.