Objective To establish an accurate sorting method with less interference to the cell activity for isolating CD 3+CD4+T lymphocytes in human intestinal tissue .Methods Firstly the digestive enzymes were adopted to decompose the human intestinal tissue ,and then the cell suspension was obtained by the filtration of 40 μm cell strainer .Mononuclear cells were collected by using continuous density Percoll gradient centrifugation technique ,then the flow cytometry was adopted to sort CD3+ CD4+ T lympho-cytes in mononuclear cells .The survival rate ,cell purity and morphological observation of the sorted cells were adopted to conduct the evaluation on the sorting method .Results The digestive enzymes well decomposed human intestinal tissue into cell suspension , the purity of CD3+CD4+ T lymphocytes by using continuous density Percoll gradient centrifugation was (43 .9 ± 7 .3)% ,while the purity of CD3+CD4+ T lymphocytes was(96 .9 ± 1 .2)% after fluorescence-activated cell sorting ,the difference between them was statistically significant(P<0 .01) .The survival rate of sorted CD3+CD4+ T lymphocytes was(97 .8 ± 1 .6)% with intact cell shape . Conclusion The Percoll gradient centrifugation technique combined with the fluorescence-activated cell sorting not only collects highly pure human intestinal CD3+CD4+ T lymphocytes with little influence on the cell activity and shape ,but aslo may be carried out under sterile conditions ,the sorted cells can continue to be used in the other functional researches .

OBJECTIVE: To study the effect of traditional chinese medicine (TCM) compound on myeloid leukemia cells and to explore its anti-leukemic mechanism. METHODS: Myeloid leukemia cell lines were cultured in vitro and treated with TCM compound. The proliferation of the leukemia cells was measured by CCK8 method. The differentiation of the leukemia cells was evaluated by using Wright＇s staining method and by light microscopy, and the expression of differentiation-related surface antigens such as CD11B was measured and by flow cytometry, the apoptosis of the leukemia cells was detected by flow cytometry with using Annexin V staining. RESULTS: Compared with untreated 4 leukemia cell lines HL-60, MOLM-13, MV4-11, AML-M5, the proliferations of 4 leukemia cells treated with different concentrations of TCM compound decreased (P<0.05), and their proliferation inhibition were in a dose-dependent manner (r=0.9236; r=0.7488; r=0.8889; r=0.8119); compared with HL-60 and AML-M5 leukemia cells, the drug-treated 2 leukemia cells displayed obvious differentiated changes; compared with untreated HL-60 leukemia cell line, the expression of surface antigen CD11B increased by 85%±7.13% in HL-60 cells treated IC50 concentration of drug; compared with untreated AML-M5 leukemia cell line, the apoptotic rate of AML-M5 treated with 1.5 and 2 μl doses of TCM compound increased. (P<0.05). CONCLUSION The traditional chinese medicine compound may inhibit the proliferation of leukemia cell lines mainly by inducing leukemia cell differentiation and apoptosis.
Apoptosis ; Cell Differentiation ; Cell Proliferation ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute

OBJECTIVE：To systematically e valuate the efficacy and safety of expectorant/antioxidants in the treatment of chronic obstructive pulmonary disease （COPD），and to provide evidence-based reference for clinical use. METHODS ：Retrieved from PubMed ，Embase，Cochrane Library ，Web of Science ，CBM，CNKI，VIP，Wanfang database ，etc.，randomized controlled trials（RCTs）about expectorant/antioxidants （trial group ）versus placebo （control group ）in the treatment of COPD were collected during the inception to May 2021. After literature screening and data extraction ，the quality of included literatures were evaluated with risk bias evaluation tool recommended by Cochrane systematic evaluator manual 5.1.0. The consistency check was performed by using Gemtc 14.3 software；network Meta-analysis ，clustering and hierarchical sorting were performed with Stata 15.1 software. The publication bias was analyzed by inverted funnel plot. RESULTS ：A total of 12 RCTs，involving 4 637 patients，were included. Five interventions measures were involved ，such as low-dose N-acetylcysteine （NAC），high-dose NAC ，carbo- cisteine， erdosteine and placebo. The results of network Meta-analysis showed that in terms of annual acute aggrava- tion rate ，the patients receiving high-do se NAC [MD ＝－0.45， 163.com 95％CI（－0.74，－0.17），P＜0.05]，carbocisteine [MD ＝－0.59，95％CI（－0.86，－0.32），P＜0.05] and erdosteine [MD ＝－0.26，95％CI（－0.51，－0.01），P＜0.05] in trial group were significantly lower than those in control group ；the annual acute aggravation rate of patients receiving high-dose NAC[MD ＝－0.55， 95％CI（－0.98，－0.11），P＜0.05] and carbocisteine [MD ＝－0.69，95％CI（－1.11，－0.26），P＜0.05] in trial group were significantly lower than those receiving low-dose of NAC ，there was no statistical significance among other groups （P＞0.05）； probability cumulative ranking results （calculated by the area under the curve ）of its network Meta-analysis was carbocisteine ＞ high-dose NAC ＞erdosteine＞placebo＞low-dose NAC. In terms of the incidence of ADR ，there was no statistical significance among groups （P＞0.05）；probability cumulative ranking results （calculated by the area under the curve ） of its network Meta-analysis was erdosteine ＞high-dose NAC ＞low-dose NAC ＞placebo＞carbocisteine. The results of clustering and hierarchical ranking showed that the efficacy and safety of the five interventions could be grouped into three categories ，including placebo and low-dose NAC with low efficacy and safety ，carbocisteine with good efficacy but low safety ，and high-dose NAC and erdosteine with good efficacy and safety. The results of publication bias showed that taking the annual acute exacerbation rate as the index ， there was a greater possibility of publication bias in this study ；taking the incidence of adverse event as index ，there was little possibility of publication bias in this study. CONCLUSIONS ：NAC，carbocisteine and erdosteine all can reduce the annual acute aggravation rate and have low incidence of ADR. Carbocisteine is the best in terms of annual acute aggravation rate ，erdosteine is the best in terms of safety. High-dose NAC and erdosteine are both better in term of efficacy and safety.

OBJECTIVETo investigate the application of the island flap based on the postfemur neurocutaneous nutrient vessel.

METHODSThe flap was designed and applied to repair the defects in the gluteal, popliteal fossa or the bilateral postfemur areas. A total of 11 cases (12 defects) were treated with this method. The size of the defects ranged from 4.0 cm x 7.8 cm to 8.3 cm x 16.6 cm.

RESULTSOf the 12 defects, 9 achieved complete success. Epidermal necrosis occurred in the distal part of the flap in 3 defects owing to venous stasis, which were cured with skin grafting. Postoperative follow-up for 8-19 months showed that the appearance, texture, and function of the flap were satisfactory.

CONCLUSIONSThe advantages of the flap lie in the reliable blood supply, constant anatomy, and without sacrificing a major artery. The key points for the flap survival are utilizing the "Superficial vein-nutrient vessel of the cutaneous nerve system" and retaining a comet tail-shaped soft-tissue pedicle in the flap creation.

Arteries ; Follow-Up Studies ; Humans ; Necrosis ; etiology ; surgery ; Skin ; injuries ; pathology ; Skin Transplantation ; Surgical Flaps ; blood supply ; pathology ; transplantation ; Thigh ; Wound Healing

Objective To investigate the role of zinc finger protein 36,C3H type-like 1 (ZFP36L1) mediating astrocytes activation in the degeneration of motor neurons in amyotrophic lateral sclerosis (ALS). Methods Superoxide dismutase 1 (S0D1)-G93A transgenic mice were used as animal models, the wild-type littermates as the control (13 mice were taken from mutant and wild-type mice at each time point) . The ZFP36L1 mRNA and protein levels of the spinal cord in the early, middle and late stage were detected by Real-time PGR and Western blotting. The expression and distribution of ZFP36L1 in the spinal cord were detected by immunofluorescence. Primary astrocyte model was established from 15 postnatal 1-2 day mice. The ZFP36L1 mRNA and protein levels in astrocytes were detected by Real-time PCR and Western blotting. Si-ZFP36L1 was transfected into SOD1-G93A mutant primary astrocytes. The transfection efficiency was detected by Western blotting. Tumor necrosis factor a (TNF-a) and interleukin-18 (IL-18) secreted from astrocytes after transfection were assessed by Western blotting and ELISA. After silencing ZFP36L1 in SOD1-G93A mutant primary astrocytes, it was cocultured with SOD1-G93A mutant NSC34 cells. 5 ' -ethynyl-2' deoxyuridine (EdU) test and the level of proliferating cell nuclear antigen (PCNA) were used to determine the effect of ZFP36L1 on NSC34 cell proliferation. TUNEL test and the level of cleaved-Caspase-3 were used to determine the effect of ZFP36L1 on NSC34 cell apoptosis. Blank small interfering RNA(siRNA) was transfected as the control group. Results Compared with the wild-type mice, the mRNA and protein levels of ZFP36L1 were downregulated in the spinal cord of SOD1-G93A transgenic mice. In wild type mice, ZFP36L1 positive cells were mainly [^-tubulin IE positive. In SOD1-G93A mutant mice, ZFP36L1 positive cells were mainly glial fibrillary acidic protein (GFAP) positive. The expression of ZFP36L1 in SOD1-G93A mutant primary astrocytes increased, and si-ZFP36Ll reduced the level of ZFP36L1 in SOD1-G93A mutant primary astrocytes significantly. Inflammatory factors including TNF-a, IL-18 decreased significantly after silencing ZFP36L1. In addition, after silencing ZFP36L1 expression, SOD1-G93A mutant primary astrocytes enhanced the proliferation activity of NSC34 cells and inhibited NSC34 cell apoptosis significantly. Conclusion Astrocytes are activated in the process of ALS. ZFP36L1 promotes the degeneration of motor neurons in ALS through the inflammatory factors secreted by astrocytes.

Objective: To analyze the epidemiological characteristics, dynamic trend of development and related influencing factors of hepatitis C in Shandong, China, 2007-2016, also to provide epidemiological evidence for prevention and control of HCV. Methods: National surveillance data of hepatitis C from 2007 to 2016 in Shandong was used, with distribution and clustering map of hepatitis C drawn at the county level. Panel Poisson regression was used to explore the influencing factors of hepatitis C at the city level. Results: The incidence of hepatitis C in Shandong increased from 1.49/100 000 in 2007 to 4.72/100 000 in 2016, with the high incidence mainly clustered in the urban regions in Jinan, Zibo, Weihai et al. and surrounding vicinities. Majority of the cases were young adults, with 53.16% (14 711/27 671) of them being farmers. Results from the Multiple panel Poisson regression analysis indicated that factors as: population density (aIRR=1.07, 95%CI: 1.05-1.10), number of hospital per hundred thousand people shared (aIRR=1.16, 95%CI: 1.08-1.24), expenditure of medical fee in rural (aIRR=1.21, 95%CI: 1.08-1.37) and the proportion of the tertiary industry (aIRR=1.08, 95%CI: 1.07-1.09) were all correlated to the incidence of hepatitis C. Conclusions: The incidence of hepatitis C had been increasing rapidly in recent years, in Shandong. Prevention and control of HCV should focus on high risk population. In addition, rural, especially in areas with lower economics provision should be under more attentions, so as to find more concealed cases for early treatment.
Adult ; China/epidemiology* ; Cities ; Hepacivirus ; Hepatitis C/prevention & control* ; Humans ; Incidence ; Population Surveillance ; Young Adult

Objective To study topological structure of a new type of three-dimensional (3D) printed height increasing insoles for leg length discrepancy (LLD) and its effect on biomechanics of lower limbs. Methods Topological structure for middle and rear part of the insole was optimized by solid isotropic microstructures with penalization (SIMP), the force was loaded and the boundary conditions were set according to force area of the insole, and the height increasing insole with thermoplastic polyurethanes (TPU) materials was printed by selected laser sintering (SLS). The insoles were used in 9 patients with LLD, visual analogue scale (VAS) and Maryland foot function scores were used to compare pain and foot function changes of patients before and after using the insole, and the 3D gait analysis system was used to compare spatiotemporal parameters and vertical ground reaction force (vGRF) of both lower limbs. Result sAfter the patient wore 3D printed insole, VAS scores decreased, Maryland foot function scores increased, vGRF of both lower limbs decreased, and the difference of cadence, stance phase and swing phase in both lower limbs decreased. Conclusions The 3D printed height increasing insole after topology optimization can improve coordination of lower limb movement, reduce ground impact, relieve pain and improve foot function, thus providing an effective personalized orthopedic plan for LLD treatment in clinic.

Objective: This study aimed to examine the demographic characteristics, HIV related knowledge and behavior, correlates of bisexual behavior and status of HIV infection among men who have sex with men only (MSMO) and men who have sex with both men and women (MSMW) in Shandong province. Methods: According to the requirements from "National HIV/AIDS sentinel surveillance program" , a cross-sectional survey was conducted to collect information on demographics, sexual and drug use behaviors, and HIV-related services among MSM in nine sentinel surveillance sites from April to July in 2018. Blood samples were drawn for serological tests on both HIV and syphilis antibodies. Results: A total of 3 474 participants were included in this study. Related information on these participants would include: average age as (31.66±9.01) years; 35.06% (1 218) married or cohabiting with a woman, 50.52% (1 755) had college or higher education, 80.11% (2 783) self-identified as gays and 14.22% (494) self-identified as bisexual men,16.87% (586) ever having sex with woman in the past 6 months, 10.51% (365) ever using drugs. HIV and syphilis prevalence rates were 2.99% (104/3 474) and 2.76%(96/3 474). Through multivariable logistic models, MSMW were more likely to be ≥35 years of age, local residents, self-identified as heterosexual/bisexual/uncertain, ever having commercial sex with man but less likely to consistently use condoms in the past 6 months, less using internet/dating software to find male sex partners and less using drugs. There was no significant differences noticed in the following areas: number of sexual partners in the last week, condom use in the last six months with commercial sex partners, with HIV or syphilis infection and self-reported history of STD in the past year between MSMO and MSMW (P>0.05). HIV-infected MSM were more likely to have the following features, ≥45 years of age, non-local residents, finding male sex partners from the bothhouses, park/toilets or from the internet/dating software, also less likely to consistently use condoms in the past 6 months, using drugs or with syphilis infection. Conclusions High prevalence of bisexual behavior as well as higher risk of HIV infection were noticed among MSM in Shandong province. It is important to strengthen related surveillance and effective intervention programs for MSM with different characteristics in Shandong province.

As a standard framework of disability definition and measurement, International Classification of Functioning, Disability and Health (ICF) is widely used in disability statistics. This paper reviewed the progress of international disability statistics based on ICF in the fields of disability measurement tools and disability surveys. There were many disability measurement tools have been developed and applied. These promote international disability statistics and disability statistics system. The disability statistics in China was analyzed and three recommendations were put forward.

Objective: To investigate the association between congenital hypothyroidism (CH) and the adverse outcomes during hospitalization in very low birth weight infants (VLBWI). Methods: This prospective, multicenter observational cohort study was conducted based on the data from the Sino-northern Neonatal Network (SNN). Data of 5 818 VLBWI with birth weight <1 500 g and gestational age between 24-<37 weeks that were admitted to the 37 neonatal intensive care units from January 1^st, 2019 to December 31^st, 2022 were collected and analyzed. Thyroid function was first screened at 7 to 10 days after birth, followed by weekly tests within the first 4 weeks, and retested at 36 weeks of corrected gestational age or before discharge. The VLBWI were assigned to the CH group or non-CH group. Chi-square test, Fisher exact probability method, Wilcoxon rank sum test, univariate and multivariate Logistic regression were used to analyze the relationship between CH and poor prognosis during hospitalization in VLBWI. Results: A total of 5 818 eligible VLBWI were enrolled, with 2 982 (51.3%) males and the gestational age of 30 (29, 31) weeks. The incidence of CH was 5.5% (319 VLBWI). Among the CH group, only 121 VLBWI (37.9%) were diagnosed at the first screening. Univariate Logistic regression analysis showed that CH was associated with increased incidence of extrauterine growth retardation (EUGR) (OR=1.31(1.04-1.64), P<0.05) and retinopathy of prematurity (ROP) of stage Ⅲ and above (OR=1.74(1.11-2.75), P<0.05). However, multivariate Logistic regression analysis showed no significant correlation between CH and EUGR, moderate to severe bronchopulmonary dysplasia, grade Ⅲ to Ⅳ intraventricular hemorrhage, neonatal necrotizing enterocolitis in stage Ⅱ or above, and ROP in stage Ⅲ or above (OR=1.04 (0.81-1.33), 0.79 (0.54-1.15), 1.15 (0.58-2.26), 1.43 (0.81-2.53), 1.12 (0.70-1.80), all P>0.05). Conclusion: There is no significant correlation between CH and in-hospital adverse outcomes, possibly due to timely diagnosis and active replacement therapy.
Infant ; Male ; Infant, Newborn ; Humans ; Female ; Prospective Studies ; Congenital Hypothyroidism/epidemiology* ; Risk Factors ; Infant, Very Low Birth Weight ; Birth Weight ; Gestational Age ; Retinopathy of Prematurity/epidemiology* ; Infant, Newborn, Diseases ; Hospitals