OBJECTIVE: To evaluate the expression level of melatonin and its effects on immune function in aplastic anemia (AA) patients. METHODS: The enzyme-linked immunosorbent assay (ELISA) was used to detect the plasma levels of melatonin in AA patients, and the correlation between melatonin levels and laboratory indexs was analyzed. The activation, proliferation, and apoptosis of T cells from AA patients were analyzed by flow cytometry with or without melatonin in vitro. RESULTS: The plasma levels of melatonin in AA patients were significantly lower compared with healthy controls (HC) (12.23 pg/ml vs 20.04 pg/ml, P < 0.01), while the plasma melatonin levels of AA patients in remission group after immunosuppressive therapy (IST) were significantly higher than those in non-remission group (29.16 pg/ml vs 11.73 pg/ml, P =0.04). Moreover, the melatonin levels were positively correlated with platelets (r =0.49), the absolute reticulocyte count (r =0.45), and the percentage of neutrophils (r =0.43). Meanwhile, there was a negative correlation between melatonin levels and the percentages of lymphocytes (r =-0.45). The expressions of CD25 and CD69 in both CD4⁺ and CD8⁺ T cells from AA patients were remarkably inhibited by melatonin in vitro (all P < 0.05). When cultured with melatonin, the proliferation rates of both CD4⁺ and CD8⁺ T cells from AA patients were markedly suppressed (P =0.01 andP < 0.01). CONCLUSION The plasma levels of melatonin were decreased in AA patients, which might play an important role in the mechanism of immunological abnormalities. The hyperimmune status of AA patients could be partially ameliorated by melatonin in vitro.
Humans ; Anemia, Aplastic ; CD8-Positive T-Lymphocytes ; Melatonin ; Blood Cell Count

Objective:To explore the difference of lymphocyte subsets in peripheral blood(PB)between aplastic anemia(AA)and hypoplastic myelodysplastic syndrome(hypo-MDS)patients,meanwhile to compare the clinical parameters obtained from PB and bone marrow(BM).Methods:The lymphocyte subsets in hypo-MDS(n=25)and AA(n=33)patients were investigated by flow cytometry.Meanwhile,the differences in PB cell counts,biochemical indicators,BM cell counts and abnormal chromosomes between the two groups were analyzed.Results:The percentage of CD8+T cells in A A group was significantly higher than that in hypo-MDS group(P=0.001),while the percentage of CD4+T cells and the CD4+/CD8+ratio in AA group were obviously lower than those in hypo-MDS group(P=0.015 and0.001,respectively).Furthermore,the proportion of CD4+andCD8+activated T(TA)cells,and memory Tregs in AA group was distinctly lower than those in hypo-MDS group(P=0.043,0.015 and 0.024,respectively).Nevertheless,the percentage of CD8+naive T(TN)cells in AA patients was remarkably higher(P=0.044).And hypo-MDS patients had declined lymphocyte counts(P=0.025),increased levels of total bilirubin(TBil),lactate dehydrogenase(LDH),vitamin B12 and proportion of BM blasts than AA patients(P=0.019,0.023,0.027 and 0.045,respectively).Conclusion:In this study it was confirmed that the percentages of CD4+and CD8+TA cells,memory Tregs and CD8+TN cells were significantly different between AA and hypo-MDS patients,which provide an essential basis for the identification of these two diseases.

Objective: To analyze the epidemiological characteristics of adult inpatients with gas burns in the Department of Burns of Hwa Mei Hospital of University of Chinese Academy of Sciences (hereinafter referred to as the author′s unit) , so as to provide evidence for the prevention of gas burn. Methods: Medical records of all inpatients with flame burns admitted to the author′s unit from January 2011 to December 2017 were collected. The percentage of adult inpatients with gas burns in total inpatients with flame burns in the same period, and their gender, age, injury season, accident place, burn severity, common compound injury, complication, population caliber, education, industry, as well as the pre-injury disease and prognosis of elderly inpatients with gas burns were retrospectively analyzed. In addition, the age, accident place, education, and industry of the floating population in the adult inpatients with gas burns were analyzed separately and compared with the total population of adult inpatients with gas burns in 7 years. Data were processed with chi-square test or Fisher′s exact probability test (Monte Carlo algorithm). Results: During the 7 years, 1 490 inpatients with flame burns were admitted to the author′s unit, among which 511 were adult inpatients with gas burns, accounting for 34.30%. The number of adult inpatients with gas burns increased gradually during the 7 years, but its percentage in the total inpatients with flame burns during the same period showed no significant difference (χ²=7.087, P>0.05). Among the 511 adult inpatients with gas burns (hereinafter referred to as the patients in this group), there were 315 males and 196 females, with a male/female ratio of 1.61 to 1.00, and the middle-aged patients were the most, up to 270 cases, accounting for 52.84%. The distribution of adult inpatients with gas burns during the 7 years was significantly different in gender and age (χ²=54.810, 27.832, P<0.01). Among the patients in this group, most were injured in summer, totally 251 cases, accounting for 49.12%, and the accident place was mainly at home, totally 388 cases, accounting for 75.93%. The distribution of adult inpatients with gas burns during the 7 years was significantly different in injury season (χ²=42.254, P<0.01), but not in accident place (χ²=6.782, P>0.05). The patients in this group were mainly with moderate burns (237 cases, accounting for 46.38%), and the distribution trend of burn severity of adult inpatients with gas burns was basically the same during the 7 years (χ²=19.680, P>0.05); 176 patients (34.44%) were accompanied by inhalation injury, and 30 patients (5.87%) were accompanied by blast injury of lung; post injury complications occurred in 20 patients (3.91%). In the elderly inpatients with gas burns, 44.44% (32/72) were accompanied by pre-injury basic diseases, and the proportion of death or unhealed reached 18.06% (13/72). Most of the patients in this group were permanent residents (358 cases, accounting for 70.06%) and received secondary education (304 cases, accounting for 59.49%), and the majority of them were engaged in manufacturing/construction (138 cases, accounting for 27.01%), self-employed business (90 cases, accounting for 17.61%), and catering (90 cases, accounting for 17.61%) industries. The distribution of adult inpatients with gas burns during the 7 years was significantly different in population caliber, education, and occupation (χ²=17.496, 29.898, 88.896, P<0.05 or P<0.01). Among the patients of this group, the floating population were mainly young (90 cases, accounting for 58.82%) and middle-aged (62 cases, accounting for 40.52%), with main accident place at home (97 cases, accounting for 63.40%), generally received secondary education (101 cases, accounting for 66.01%), and were mainly engaged in manufacturing/construction (71 cases, accounting for 46.41%), self-employed business (26 cases, accounting for 16.99%), and catering (20 cases, accounting for 13.07%) industries. Compared with the total adult inpatients with gas burns in 7 years, the floating population were younger, more injured in the workplace, and more concentrated in industry (χ²=42.924, 9.390, 27.819, P<0.01). Conclusions Gas burn was the leading injury cause of inpatients with flame burns in the author′s unit, which mainly occurred in summer and at home; the patients were mainly male, young and middle-aged, and permanent residents, most of which were with moderate burn, often accompanied by inhalation injury. Most of the patients were of secondary education, engaged in manufacturing/construction, self-employed business, and catering industries, among which the floating population were younger, more injured in the workplace, and more concentrated in industry. In order to prevent gas burn, we should pay more attention to the propaganda and education of gas safety among young and middle-aged men, floating population, retired old people and housewives, especially in summer, we should do a good job in gas safety inspection at home. In addition, we should urge enterprises to further strengthen the supervision of production safety.

From June to November 2016, 5 patients with severe burns were admitted to our unit. Broad-spectrum antibiotic and fluconazole were used on patients as earlier empirical anti-infection therapy of bacteria and fungi. Seven to twenty-one days after injury, 5 patients developed fungal infection. Antifungal agents of caspofungin, voriconazole, and amphotericin B liposomewere were used according to the results of fungal culture, and the infected wounds were also treated with repeated debridement and dressing change. Multiple autologous skin grafts were performed after infection control of wounds. With the above antifungal infection treatment for 5 to 11 days, 2 patients′ condition tended to be stable, and no fungus was found in wound secretion after cultured for many times. The patients were discharged with wounds healed after 52 to 54 days′ hospital stay. Due to severe burns degree and or elder age, fungal infection aggravated and expanded to the trunk in the other 3 patients, then developed into burn sepsis, resulting in patients died of multiple organ failure secondary to sepsis.

Objective: To investigate the effect of calcitonin (CT) on promoting collagen synthesis and osteogenesis of human periodontal ligament stem cells (hPDLSCs). Methods: Fifty adult participants were divided into chronic periodontitis (CP) group (n=25) and control group (n=25). In the CP group, the anterior maxilla with probing depth ≥5 mm and the sites with imaging evidence of bone loss were selected. The gingival crevicular fluid (GCF) samples were collected from 6 maxillary sites in each patient. In the control group, multiple sites without inflammation (10 to 12 per subject) were sampled to ensure that a sufficient amount of GCF was collected. The expression of CT, transforming growth factor β1 (TGF-β1) and bone morphogenetic protein (BMP) 2/4/7 in GCF was detected by enzyme linked immunosorbent assay (ELISA), and the correlation between CT expression and clinical parameters such as periodontal pocket probing depth (PD), clinical attachment loss (CAL) and gingival index (GI), and the above-mentioned indicators was investigated with Spearman correlation analysis. hPDLSCs were infected with the adenoviruses carrying CT gene (Ad.CT) and the expression of mRNA and protein of TGF-β1, BMP2/4/7, alkaline phosphatase (ALP), osteocalcin (OCN) and collagen type I/III (Col I/III) were detected by quantitative real-time PCR and Western blotting. Results: The expression level of CT in GCF of the CP group was significantly higher than that of the control group ([32.62±1.46] ng/mL vs [17.70 ± 0.76] ng/mL, P<0.01). The expression of CT was positively correlated with clinical parameters such as PD, CAL and GI (P<0.01, P<0.05). The expression levels of BMP2/4/7 and TGF-β1 in GCF of the CP group were significantly higher than those of the control group (BMP2: [138.67 ± 4.04] ng/mL vs [103.96 ± 2.78] ng/mL, BMP4: [155.53 ± 3.55] ng/mL vs [133.15 ± 2.92] ng/mL; BMP7: [106.59 ± 2.85] ng/mL vs [90.22±1.56] ng/mL; TGF-β1: [105.92 ± 3.40] ng/mL vs [89.85 ± 2.42] ng/mL; all P<0.01). The expression of BMP2/4/7 and TGF-β1 was negatively correlated with CT expression (P<0.01, P<0.05). The overexpression of CT significantly increased the expression of TGF-β1, Col I/III and osteoblast markers BMP2/4, ALP and OCN in GCF (all P<0.01). Compared with the cells co-infected with Ad.CT and Ad.Null, the cells co-infected with Ad.CT and small interfering RNA specifically blocking TGF-β1 (Ad.TGF-β1 siRNA) had significantly lower collagen expression (Col I: 0.16 ± 0.02 vs 0.22 ± 0.03; Col III: 0.11±0.01 vs 0.15 ± 0.02; both P<0.01). Compared with Ad.CT infected cells, the protein expression levels of ALP and OCN were significantly decreased in Ad.CT and noggin co-treated cells (ALP: 0.19 ± 0.02 vs 0.25 ± 0.03; OCN: 0.13 ± 0.01 vs 0.19 ± 0.02; both P<0.01). Conclusion: CT can promote collagen synthesis and osteogenesis in hPDLSCs through TGF-β1 and BMP signaling transduction pathways.

Objective: To analyze clonal evolution and clinical significance of trisomy 8 in patients with acquired bone marrow failure. Methods: The clinical data of 63 patients with acquired bone marrow failure accompanied with isolated trisomy 8 (+8) from June 2011 to September 2018 were analyzed retrospectively, the clonal evolution patterns and relationship with immmunosuppressive therapy were summarized. Results: Totally 24 male and 39 female patients were enrolled, including 39 patients with aplastic anemia (AA) and 24 patients with relatively low-risk myelodysplastic syndrome (MDS) . Mean size of+8 clone in MDS patients[65% (15%-100%) ]was higher than that of AA patients[25% (4.8%-100%) , z=3.48, P=0.001]. The patients were was divided into three groups (<30%, 30%-<50%,and ≥50%) according to the proportion of+8 clone. There was significant difference among the three groups between AA[<30%:55.6% (20/36) ; 30-50%: 22.2% (8/36) ; ≥50%22.2% (8/36) ]and MDS patients[<30%:19.0% (4/21) ; 30%-<50%:19.0% (4/21) ; ≥50%61.9% (13/21) ] (P=0.007) . The proportion of AA patients with+8 clone <30% was significantly higher than that of MDS patients (P=0.002) ; and the proportion of AA patients with+8 clone ≥50%was significantly lower than that of MDS patients (P=0.002) . The median age of AA and MDS patients was respectively 28 (7-61) years old and 48.5 (16-72) years old. Moreover, there was no correlation between age and+8 clone size in AA or MDS (r(s)=0.109, P=0.125; r(s)=-0.022, P=0.924, respectively) . There was statistical difference in total iron binding capacity, transferrin and erythropoietin between high and low clone group of AA patients (P=0.016, P=0.046, P=0.012, respectively) , but no significant difference in MDS patients. The immunosuppressive therapy (IST) efficacy of AA and MDS patients was respectively 66.7% and 43.8% (P=0.125) . Comparing with initial clone size (27.3%) , the +8 clone size (45%) of AA patients was increased 1-2 year after IST, but no statistical difference (z=0.83, P=0.272) . Consistently, there was no significant change between initial clone size (72.5%) and 1-2 year clone size (70.5%) after IST in MDS patients. There was no significant difference in IST efficient rate between +8 clone size expansion and decline group of in AA patients at 0.5-<1, 1-2 and>2 years after IST. We found four dynamic evolution patterns of +8 clone, which were clone persistence (45%) , clone disappearance (30%) , clone emergence (10%) and clone recurrence (15%) . Conclusions: AA patients had a low clone burden, while MDS patients had a high burden of +8 clone. The +8 clone of AA patients didn't significantly expanded after IST, and the changes of +8 clone also had no effect on IST response.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; Bone Marrow ; Child ; Chromosomes, Human, Pair 8 ; Clonal Evolution ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Trisomy ; Young Adult

OBJECTIVETo detect the expression of miRNA in de novo and complete response SAA patients and predict the targets of the miRNAs.

METHODSThe expression profiles of miRNA from bone marrow mononuclear cells of the SAA patients with de novo and CR were detected by miRNA microarray.

RESULTSTotally 35 up-regulated and 37 down-regulated miRNA were identified in CR SAA patients in comparison with de novo SAA patients. Furthermore, by predicting the targets of the differentlly expressed miRNA, it was found that some targets associated with T cell receptor signaling pathway and cell adhesion molecules.

CONCLUSIONSome miRNA may be involved in the pathogenesis of SAA.

Objective: To determine the valuable hemolytic characteristics in differential diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), autoimmune hemolytic anemia (AIHA) and hereditary spherocytosis (HS). Method: The clinical and hemolytic characteristics of 108 PNH patients, 127 AIHA patients and 172 HS patients diagnosed from January 1998 to April 2017 were compared. Results: ①Reticulocyte percentage (Ret%) of PNH patients [6.70% (0.14%-22.82%)] was significantly lower than that of AIHA [14.00%(0.10%-55.95%), P<0.001] and HS patients [11.83%(0.60%-57.39%), P<0.001]. The Ret% in PNH patients were significantly lower than those in AIHA and HS patients at the same levels of anemia, except for in mild anemia between PNH and AIHA patients. However, when comparing the Ret% between AIHA and HS patients, there was significant difference only in mild anemia [7.63%(1.87%-29.20%)% vs 11.20%(3.31%-22.44%), z=-2.165, P=0.030]. ②The level of TBIL in HS patients was significantly higher than that in AIHA and PNH patients [79.3 (11.2-244.0) μmol/L vs 57.6 (7.6-265.0) μmol/L, z=5.469, P<0.001; 79.3(11.2-244.0) μmol/L vs 26.2(4.6-217.7) μmol/L, z=-2.165, P<0.001], and the proportion of HS patients with TBIL more than 4 times the upper limit of normal (ULN) (64.1%) was significantly higher than that of AIHA (37.7%, χ(2)=19.896, P<0.001) and PNH patients (4.6%, P<0.001). ③The LDH level of PNH patients was significantly higher than that of AIHA and HS [1 500 (216-5 144) U/L vs 487 (29-3 516) U/L, z=-9.556, P<0.001; 1 500 (216-5 144) U/L vs 252 (132-663) U/L, z=-11.518, P<0.001], and the proportion of PNH patients with LDH more than 1 000 U/L (79.1%) was significantly higher than that of AIHA patients (13.0%, χ(2)=93.748, P<0.001) and HS patients (0, P<0.001). ④Splenomegaly occurred in 43.5% of PNH patients, including 16.0% with severe splenomegaly. In contrast, the occurrence of splenomegaly was 98.6% in AIHA patients and 100.0% in HS patients (P<0.001), and 63.0% of AIHA patients (P<0.001) and 90.4% of HS patients (P<0.001) were with severe splenomegaly. ⑤The prevalence of cholelithiasis in HS patients was up to 43.1%, significantly higher than that in AIHA patients (10.5%, P<0.001) and PNH patients (2.9%, P<0.001). Conclusion: The comprehensive assessment of the five hemolytic characteristics is simplified, practical and efficient, with great clinical significance, providing specific indicators for differential diagnosis and efficient approach for making further work-up.
Anemia, Hemolytic, Autoimmune ; Diagnosis, Differential ; Hemoglobinuria, Paroxysmal ; Hemolysis ; Humans ; Spherocytosis, Hereditary

OBJECTIVETo explore the clinical characteristic, therapeutic efficacy and prognosis of patients with hepatitis-associated aplasitc anemia (HAAA).

METHODSthe clinical data and labrotatory examination results of 30 cases of HAAA were analyzed retrospectively, the 6-month response ratio and overall survival (OS) were assessed.

RESULTSHAAA most commonly occured in males, with the occurence rate of males and females was 4:1, the median onset age was 16 (4-43) years old, HAAA oriented focus on sever aplastic anemia (SAA)(4 cases,13%) and very sever aplastic anemia (VSAA)(22 cases,73%). Aplastic anemia (AA) could be seen on occurence of hepatitis (accompanied aplastic anemia) (7 cases,23%), or after the onset of hepatits (delayed aplastic anemia) (23 cases,77%), but more often occured in the latter. Statistical analysis showed that when compared with the patients of delayed aplastic anemia, patients accompanied aplastic anemia possesses lower levels of glutamic-pyruvic transaminase(ALT), aspertate aminotransferase (AST) and total bilirubin (TBIL)(P=0.042,0.012,0.001), and possessed a more obvious lymphoid cell disorder when AA occured, with more lower peripheral blood CD19B cells proportion (P=0.046) and more obvious imbalance of CD4/CD8ratio, but the difference was no statistical significant (P=0538). Factors affecting the 6-month respose were the severity of AA (P=0.044), the peak level of bilirubin of hepatitis (P=0.006) and the propotion of mature monocyte in bone marrow (P=0.034). The long-term follow-up showed that the 2-year OS of HAAA was 64.3±9.2%, the 6-month curative efficacy significantly affect the prognosis (P<0.001).

CONCLUSIONHAAA more often occur in young male, HAAA is mainly SAA and VSAA and mostly non-A-C hepatitis associated aplastic anemia, patients usually have a high incidence of early infection. Patients acompanied with aplastic anemia possess more obvious immunological derangement; the treatment efficacy for HAAA is poor, patients who haven't obtained 6-month response indicate a sinister prognosis, allogeneic hematopoietic stem cell transplantion is a better choice for these patients.