·AIM:To discuss the clinical efficacy of 27-gauge (27G) vitrectomy with proliferative membrane cutting in situ for late proliferative diabetic retinopathy (PDR). ·METHODS: Collecting 10 cases (15 eyes) with late PDR from January 2017 to August 2017 which underwent 27G microincision vitrectomy with cutting proliferative membrane in situ, we observed the rate of intraoperative iatrogenic retinal hole ( IRH ), the rate of silicone oil tamponade, the best corrected visual acuity ( BCVA) and intraocular pressure (IOP) before and after operation. ·RESULTS: IRH occurred in 4 eyes (27%, 4/15); Silicone oil was tamponaded in 6 eyes (40%, 6/15); BCVA was improved in 13 eyes and only 2 eyes unchanged 3mo after operation. The best visual acuity (VA) was 0. 6. There was significant difference on BCVA between preoperative and postoperative 7d(P<0. 05). The same was found between preoperative and postoperative 1mo, even 3mo(P<0. 05). The average preoperative IOP was 16. 95 ± 6. 87mmHg and postoperative 3mo was 15. 27 ± 4. 57mmHg. There was no significant difference between them (P>0. 05). · CONCLUSION: The 27G vitrectomy with cutting proliferative membrane in situ method is markedly superior in the treatment of late PDR, and the curative effect is specific. It can be given preference to late PDR.

OBJECTIVETo investigate the prevalence of JAK2V617F and MPLW515L/K mutation in patients with slightly elevated platelets (BPC) or hemoglobin (Hb) not meeting the criteria of polycythemia vera (PV) or essential thrombocythemia (ET).

METHODSGenomic DNA from bone marrow or blood mononuclear cells was screened with allele specific polymerase chain reaction (AS-PCR) for JAK2V617F and MPLW515L/K mutation. The history of thrombosis was assessed retrospectively by patients files.

RESULTSOf 30 patients, 14 (46.7%) were positive for the JAK2V617F mutation, none of them had the MPLW515L/ K. Five of these 14 patients had a history of thrombosis. Follow-up results were available in 22 patients. Among them, 12 patients with JAK2V617F mutation turned out to be MPD in 6-24 months; only 2 out of 10 patients without this mutation evolved to MPD.

CONCLUSIONJAK2V617F mutation could be one of the diagnosis criteria of early MPD. No MPLW515L/K expression was found in early MPD.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Early Diagnosis ; Female ; Follow-Up Studies ; Humans ; Janus Kinase 2 ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; diagnosis ; genetics ; metabolism ; Receptors, Thrombopoietin ; genetics ; metabolism ; Young Adult

The aim was to investigate the cytolytic activity of cytotoxic T lymphocytes (CTL) induced by dendritic cells (DC) derived from human cord blood in vitro. Human cord blood mononuclear cells (CBMNC) were cultured in vitro with addition of various cytokines. DC was confirmed by morphology, immune phenotype and capacity of stimulating MLR (mixed lymphocyte reaction). CTL were generated through the co-culture of autologous T lymphocytes and DC. (51)Cr-release assays were performed for the measurement of cytotoxicity of CTL. The results showed that distribution of the subgroups of T lymphocytes in CBMNC was similar to that in adult peripheral blood. The percentage of CD1a-expressing cells in CBMNC was very low, merely (0.41 +/- 0.09)%. During culture, DC became larger and more irregular in shape. Spiny dendrites or multiple cell processes in morphology emerged on the surface of DC. Among the cell populations at 15 days of culture, there were (28.4 +/- 3.55)% of CD1a-expressing cells, (63.67 +/- 23.33)% of CD86-positive, (8.7 +/- 1.49)% of CD83-positive and (32.5 +/- 1.53)% of HLA-DR-positive cells. DC derived from CBMNC is capable of stimulating the proliferation of allogeneic lymphocytes in MLR. CTL derived from autologous T lymphocytes induced by dendritic cells pulsed with lysates of HL-60 cells, possessed specific cytolytic effects against HL-60 cells. In conclusions, relatively high percentage of CD1a-expressing cells can be generated in culture system of this study. DC derived from cord blood is able to induce the production of anti-leukemic CTL in vitro.
Antigens, CD1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Fetal Blood ; immunology ; Humans ; Immunophenotyping ; Leukemia ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.

METHODSHUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.

RESULTSs The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.

CONCLUSIONDigestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.

Cell Culture Techniques ; Cell Cycle ; Cells, Cultured ; Culture Media ; chemistry ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 8 ; chemistry ; Serum

OBJECTIVETo analyze the relationship between the reference values of fibrinogen (FIB) in healthy Chinese adults and geographical factors to provide scientific evidences for establishing the uniform standard.

METHODSThe reference values of FIB of 10701 Chinese healthy adults from 103 cities were collected to investigate their relationship with 18 geographical factors including spatial index, terrain index, climate index, and soil index. Geographical factors that significantly correlated with the reference values were selected for constructing the BP neural network model. The spatial distribution map of the reference value of FIB of healthy Chinese adults was fitted by disjunctive kriging interpolation. We used the 5-layer neural network and selected 2000 times of training covering 11 hidden layers to build the simulation rule for simulating the relationship between FIB and geographical environmental factors using the MATLAB software.

RESULTSs The reference value of FIB in healthy Chinese adults was significantly correlated with the latitude, sunshine duration, annual average temperature, annual average relative humidity, annual precipitation, annual range of air temperature, average annual soil gravel content, and soil cation exchange capacity (silt). The artificial neural networks were created to analyze the simulation of the selected indicators of geographical factors. The spatial distribution map of the reference values of FIB in healthy Chinese adults showed a distribution pattern that FIB levels were higher in the South and lower in the North, and higher in the East and lower in the West.

CONCLUSIONWhen the geographical factors of a certain area are known, the reference values of FIB in healthy Chinese adults can be obtained by establishing the neural network mode or plotting the spatial distribution map.

Adult ; Asian Continental Ancestry Group ; China ; Climate ; Environment ; Fibrinogen ; Geography ; Humans ; Neural Networks (Computer) ; Reference Values ; Software ; Temperature

BACKGROUNDThe cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro.

METHODSFluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT).

RESULTSThe molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group.

CONCLUSIONSThe newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.

Cell Line, Tumor ; Cell Membrane Permeability ; Contrast Media ; Fluorescein-5-isothiocyanate ; Gadolinium DTPA ; Humans ; Magnetic Resonance Imaging ; methods ; Peptides ; metabolism

Objective To investigate the clinical characteristics and therapeutic methods of bac-terial colonization and infection of Acinetobacter baumannii on the wound of earthquake induced patients. Methods A retrospective study was done on 42 Wenehuan earthquake induced patients with positive wound germiculture of Acinetobacter baumannii. There were 24 males and 18 females, at mean age of 37 years (12-96 years). Open injury was located at the upper arm in one patient, at the forearm in four, at the thigh in 12, at the calf in 23 and at the trunk in two. The time between injury and treatment varied from 3 to 7 days. The clinical characteristics including the bacteria identification and drug sensitivity test were studied to compare drug resistance to 15 antibiotics. Results Bacterial colonization of Acineto-bacter baumannii was found in 31 patients (8.2%) and infection of Acinetobacter baumannii in 11 (2.9%). After debridement, pruphylactic antibiotics and nutrition support, 15 patients with bacterial colonization were managed with Ⅱ stage suture or skin grafting. The other 16 patients were transferred to hospitals of other provinces after germiculture turned negative. Through debridement and drainage, antibi-otic therapy and nutrition support, the infection was controlled and the wound eliminated in six patients through Ⅱ stage suture but four were concomitant with pulmonary infection and one with septicemia. Drug sensitivity test showed that sensitive rate to imipenem, amikacin, levofloxacin, ticarcillin-clavulanic acid, tobramycin were 59.5%, 21.4%, 21.4%, 19.5% and 19.0% respectively. Conclusions The risk factors of infection of Acinetobacter baumannii include severe tissue trauma, severe wound contamination, delayed treatment and weak body resistance. During treatment, the bacterial colonization and infection of Acinetobacter baumannii should be distinguished and treated respectively. Correct wound treatment, suit-able antibiotic therapy and increased body resistance are key to improvement of clinical curative effect.

BACKGROUNDProteasome subunits (PSMB) and transporter associated with antigen processing (TAP) loci are located in the human leukocyte antigen (HLA) Class II region play important roles in immune response and protein degradation in neurodegenerative diseases. This study aimed to explore the association between single nucleotide polymorphisms (SNPs) of PSMB and TAP and Parkinson's disease (PD).

METHODSA case-control study was conducted by genotyping SNPs in PSMB8, PSMB9, TAP1, and TAP2 genes in the Chinese population. Subjects included 542 sporadic patients with PD and 674 healthy controls. Nine identified SNPs in PSMB8, PSMB9, TAP1, and TAP2 were genotyped through SNaPshot testing.

RESULTSThe stratified analysis of rs17587 was specially performed on gender. Data revealed that female patients carry a higher frequency of rs17587-G/G versus (A/A + G/A) compared with controls. But there was no significant difference with respect to the genotypic frequencies of the SNPs in PSMB8, TAP1, and TAP2 loci in PD patients.

CONCLUSIONChinese females carrying the rs17587-G/G genotype in PSMB9 may increase a higher risk for PD, but no linkage was found between other SNPs in HLA Class II region and PD.

ATP-Binding Cassette Sub-Family B Member 2 ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; genetics ; Adult ; Aged ; Antigen Presentation ; Case-Control Studies ; Cysteine Endopeptidases ; genetics ; Female ; Humans ; Male ; Middle Aged ; Parkinson Disease ; genetics ; immunology ; Polymorphism, Single Nucleotide ; Proteasome Endopeptidase Complex ; genetics

Objective:To compare the clinical efficacy between femoral neck system (FNS) and inverted cannulated compression screws (ICCS) in the fixation of adult femoral neck fracture.Methods:The clinical data were retrospectively analyzed of the 119 patients with femoral neck fracture who had received FNS or ICCS internal fixation at Department of Traumatic Orthopedics, West China Hospital from September 2019 to June 2020. They were divided into 2 groups according to their internal fixation methods. In the FNS group of 62 patients, there were 38 males and 24 females, with an age of (54.0±13.0) years, and 13 cases of type Ⅱ, 34 cases of type Ⅲ and 15 cases of type Ⅳ according to the Garden classification; in the ICCS group of 57 patients, there were 42 males and 15 females, with an age of (53.2±11.3) years, and 9 cases of type Ⅱ, 33 cases of type Ⅲ and 15 cases of type Ⅳ according to the Garden classification. The operation time, intraoperative blood loss, fluoroscopy frequency, hospitalization time, fracture healing time, Harris hip score and incidence of complications were compared between the 2 groups.Results:The 2 groups were comparable due to insignificant differences in their preoperative general data or follow-up duration ( P>0.05). There were significant differences between the FNS and ICCS groups in fluoroscopy frequency [(8.8±2.9) times versus (15.6±3.4) times], operation time [(45.2±10.1) min versus (51.8±11.7) min], fracture healing time [(3.2±0.4) months versus (4.0±0.6) months], Harris hip score at the last follow-up [(91.8±4.4) points versus (84.6±3.3) points], and femoral neck shortening at the last follow-up, favoring the FNS group (all P<0.05). There were no significant differences in follow-up time, hospitalization time, intraoperative blood loss or incidence of complications between the 2 groups ( P>0.05). Conclusions:In the fixation of adult femoral neck fractures, compared with ICCS, FNS can significantly reduce fluoroscopy frequency, shorten fracture healing and operation time, reduce risk of femoral neck shortening and hospitalization time, and promote functional recovery of the hip.

BACKGROUNDNurr1 plays an essential role in the development, survival, and function maintenance of midbrain dopaminergic (DA) neurons, and it is a potential target for Parkinson's disease (PD). Nurr1 mRNA can be detected in peripheral blood mononuclear cells (PBMCs), but whether there is any association of altered Nurr1 expression in PBMC with the disease and DA drug treatments remains elusive. This study aimed to measure the Nurr1 mRNA level in PBMC and evaluate the effect of Nurr1 expression by DA agents in vivo and in vitro.

METHODSThe mRNA levels of Nurr1 in PBMC of four subgroups of 362 PD patients and 193 healthy controls (HCs) using real-time polymerase chain reaction were measured. The nonparametric Mann-Whitney U-test and Kruskal-Wallis test were performed to evaluate the differences between PD and HC, as well as the subgroups of PD. Multivariate linear regression analysis was used to evaluate the independent association of Nurr1 expression with Hoehn and Yahr scale, age, and drug treatments. Besides, the Nurr1 expression in cultured PBMC was measured to determine whether DA agonist pramipexole affects its mRNA level.

RESULTSThe relative Nurr1 mRNA levels in DA agonists treated subgroup were significant higher than those in recent-onset cases without any anti-PD treatments (de novo) (P < 0.001) and HC groups (P < 0.010), respectively. Furthermore, the increase in Nurr1 mRNA expression was seen in DA agonist and L-dopa group. Multivariate linear regression showed DA agonists, L-dopa, and DA agonists were independent predictors correlated with Nurr1 mRNA expression level in PBMC. In vitro, in the cultured PBMC treated with 10 μmol/L pramipexole, the Nurr1 mRNA levels were significantly increased by 99.61%, 71.75%, 73.16% in 2, 4, and 8 h, respectively (P < 0.001).

CONCLUSIONSDA agonists can induce Nurr1 expression in PBMC, and such effect may contribute to DA agonists-mediated neuroprotection on DA neurons.

Adult ; Aged ; Aged, 80 and over ; Dopamine Agonists ; therapeutic use ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Middle Aged ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; genetics ; Parkinson Disease ; drug therapy ; genetics ; RNA, Messenger ; genetics ; Young Adult