1.Efficacy of electrolysis of depigmented hair combined with hair follicle transplantation in the treatment of 25 cases of vitiligo-associated leukotrichia: a retrospective analysis
Tao WANG ; Yi WU ; Hong JIN ; Yeqin DAI ; Jianzhong PENG ; Ai′e XU ; Xiuzu SONG
Chinese Journal of Dermatology 2024;57(1):46-49
		                        		
		                        			
		                        			Objective:To analyze the clinical efficacy of electrolysis of depigmented hair using a trichiasis electrolyzer combined with hair follicle transplantation in the treatment of vitiligo-associated leukotrichia.Methods:Clinical data were retrospectively collected from 25 patients with stable vitiligo-associated leukotrichia in the Department of Dermatologic Surgery, Hangzhou Third People′s Hospital from January 2019 to January 2021. All the patients received electrolysis of depigmented hair using a trichiasis electrolyzer combined with hair follicle transplantation. Outpatient follow-up visits were conducted in the first week, as well as the first, third and sixth months after surgery. The texture and growth status of transplanted hair were observed, and the survival rate of transplanted hair follicles and the proportion of newborn white hair in white hair in the original lesions were recorded.Results:Among the 25 patients with stable vitiligo, there were 14 males and 11 females, and their disease duration ranged from 2 to 15 years, with the average duration being 5.8 years. A total of 30 white patches accompanied by leukotrichia were included, including 9 on the scalp, 7 on the eyebrows and 14 on the eyelashes. One week after surgery, the transplanted hair survived well in all patients, without obvious shedding or local infection. Six months after surgery, repigmentation was observed in most hair in the original lesion area, and only a small amount of white hair grew out, without obvious scarring; the survival rate of transplanted hair follicles was 76.5% ± 10.0%, and the proportion of newborn white hair in white hair in the original lesions was 16.7% ± 7.8%.Conclusion:Electrolysis of depigmented hair using a trichiasis electrolyzer combined with hair follicle transplantation was effective in the treatment of vitiligo-associated leukotrichia, with a simple treatment process and few postoperative complications, which provided a reliable choice for the clinical treatment of vitiligo-associated leukotrichia.
		                        		
		                        		
		                        		
		                        	
2.Expression and Clinical Significance of Exosome Derived MiR-181b-5p in Children with Acute Lymphoblastic Leukemia.
Yi HONG ; Kang-Kang LIU ; Ning-Ling WANG ; Zhi-Wei XIE ; Jin-Hua CHU
Journal of Experimental Hematology 2023;31(3):643-648
		                        		
		                        			OBJECTIVE:
		                        			To explore the expression level of exosome derived miR-181b-5p in different disease stages of children with acute lymphoblastic leukemia and its relationship with clinical characteristics.
		                        		
		                        			METHODS:
		                        			Bone marrow plasma samples of 86 children with ALL were collected. Exosomes were extracted by exosome extraction kit, and RNA in exosomes was extracted by TRIzol method. The levels of miR-181b-5p in the blood plasma exosomes of the patients in the newly diagnosed group, relapse group, remission group and control group were detected by qRT- PCR. The difference of miR-181b-5p expression level in each group was compared and analyzed, and the relationship between miR-181b-5p expression level and clinical characteristics was analyzed.
		                        		
		                        			RESULTS:
		                        			The expression level of exosomal miR-181b-5p in the newly diagnosed group and the relapsed group was significantly lower than that in the remission group and the control group (P< 0.05). The expression level of exosomal miR-181b-5p in T-ALL children was higher than that in B-ALL children (P<0.05). The expression level of plasma exosomal miR-181b-5p in male children was higher than that in female children (P<0.01).
		                        		
		                        			CONCLUSION
		                        			Exosome derived miR-181b-5p changes dynamically in the course of ALL children, and can be used as a marker miRNA to monitor disease status. Exosomes can transmit information in the tumor microenvironment and serve as a potential carrier for biomolecular targeted therapy.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Child
		                        			;
		                        		
		                        			Exosomes/metabolism*
		                        			;
		                        		
		                        			Clinical Relevance
		                        			;
		                        		
		                        			MicroRNAs/genetics*
		                        			;
		                        		
		                        			Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism*
		                        			;
		                        		
		                        			Tumor Microenvironment
		                        			
		                        		
		                        	
3.Distribution and Drug Resistance of Pathogens in Oral Mucositis Associated with Chemotherapy in Patients with Malignant Hematopathy.
Jin QIU ; Zi-Hao ZHANG ; Xiao-Ting LIU ; Cheng-Long LIU ; Si-Yi ZHU ; Zhao-Qu WEN
Journal of Experimental Hematology 2023;31(1):274-279
		                        		
		                        			OBJECTIVE:
		                        			To analyze the distribution and drug resistance of pathogens in oral mucositis associated with chemotherapy in hospitalized patients with malignant hematopathy, so as to provide scientific evidences for rational selection of antibiotics and infection prevention and control.
		                        		
		                        			METHODS:
		                        			From July 2020 to June 2022, 167 patients with malignant hematopathy were treated with chemical drugs in the Department of Hematology, Hainan Hospital, and secretions from oral mucosal infected wounds were collected. VITEK2 COMPECT automatic microbial identification system (BioMerieux, France) and bacterial susceptibility card (BioMerieux) were used for bacterial identification and drug susceptibility tests.
		                        		
		                        			RESULTS:
		                        			A total of 352 strains of pathogens were isolated from 167 patients, among which 220 strains of Gram-positive bacteria, 118 strains of Gram-negative bacteria and 14 strains of fungi, accounted for 62.50%, 33.52% and 3.98%, respectively. The Gram-positive bacteria was mainly Staphylococcus and Streptococcus, while Gram-negative bacteria was mainly Klebsiella and Proteus. The resistance of main Gram-positive bacteria to vancomycin, ciprofloxacin and gentamicin was low, and the resistance to penicillin, cefuroxime, ampicillin, cefotaxime, erythromycin and levofloxacin was high. The main Gram-negative bacteria had low resistance to gentamicin, imipenem and penicillin, but high resistance to levofloxacin, cefotaxime, cefuroxime, ampicillin and vancomycin. The clinical data of oral mucositis patients with oral ulcer (severe) and without oral ulcer (mild) were compared, and it was found that there were statistically significant differences in poor oral hygiene, diabetes, sleep duration less than 8 hours per night between two groups (P<0.05).
		                        		
		                        			CONCLUSION
		                        			Gram-positive bacteria is the main pathogen of oral mucositis in patients with malignant hematopathy after chemotherapy. It is sensitive to glycopeptide antibiotics and aminoglycosides antibiotics. Poor oral hygiene, diabetes and sleep duration less than 8 hours per night are risk factors for oral mucositis with oral ulcer (severe).
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Vancomycin/therapeutic use*
		                        			;
		                        		
		                        			Cefuroxime
		                        			;
		                        		
		                        			Levofloxacin
		                        			;
		                        		
		                        			Oral Ulcer/drug therapy*
		                        			;
		                        		
		                        			Drug Resistance, Bacterial
		                        			;
		                        		
		                        			Anti-Bacterial Agents/adverse effects*
		                        			;
		                        		
		                        			Ampicillin
		                        			;
		                        		
		                        			Penicillins
		                        			;
		                        		
		                        			Cefotaxime
		                        			;
		                        		
		                        			Gram-Positive Bacteria
		                        			;
		                        		
		                        			Gram-Negative Bacteria
		                        			;
		                        		
		                        			Gentamicins
		                        			;
		                        		
		                        			Stomatitis/drug therapy*
		                        			
		                        		
		                        	
4.Clinical analysis of 86 cases of acute fatty liver of pregnancy.
Cong Li LIU ; Dun Jin CHEN ; Chu Yi CHEN ; Xiu Hua ZHOU ; Yuan JIANG ; Jing Yu LIU ; Yue E CHEN ; Cheng Ran HU ; Jin Ju DONG ; Ping LI ; Min WEN ; Yan Hui LI ; Hui Li ZHANG
Chinese Journal of Obstetrics and Gynecology 2023;58(12):896-902
		                        		
		                        			
		                        			Objective: To investigate the clinical characteristics and maternal and fetal prognosis of pregnant women with acute fatty liver of pregnancy (AFLP). Methods: The clinical data of 86 AFLP pregnant women admitted to the Third Affiliated Hospital of Guangzhou Medical University from September 2017 to August 2022 were collected, and their general data, clinical characteristics, laboratory tests and maternal and fetal outcomes were retrospectively analyzed. Results: (1) General information: the age of the 86 pregnant women with AFLP was (30.8±5.4) years, and the body mass index was (21.0±2.5) kg/m2. There were 50 primiparas (58.1%, 50/86) and 36 multiparas (41.9%, 36/86). There were 64 singleton pregnancies (74.4%, 64/86) and 22 twin pregnancies (25.6%, 22/86). (2) Clinical characteristics: the main complaints of AFLP pregnant women were gastrointestinal symptoms, including epigastric pain (68.6%, 59/86), nausea (47.7%, 41/86), anorexia (46.5%, 40/86), vomiting (39.5%, 34/86). The main non-gastrointestinal symptoms were jaundice of skin and/or scleral (54.7%, 47/86), edema (38.4%, 33/86), fatigue (19.8%, 17/86), bleeding tendency (16.3%, 14/86), polydipsia or polyuria (14.0%, 12/86), skin itching (8.1%, 7/86), and 17.4% (15/86) AFLP pregnant women had no obvious symptoms. (3) Laboratory tests: the incidence of liver and kidney dysfunction and abnormal coagulation function in AFLP pregnant women was high, and the levels of blood ammonia, lactate dehydrogenase and lactic acid were increased, and the levels of hemoglobin, platelet and albumin decreased. However, only 24 cases (27.9%, 24/86) of AFLP pregnant women showed fatty liver by imageology examination. (4) Pregnancy outcomes: ① AFLP pregnant women had a high incidence of pregnancy complications, mainly including renal insufficiency (95.3%, 82/86), preterm birth (46.5%, 40/86), hypertensive disorders in pregnancy (30.2%, 26/86), gestational diabetes mellitus (36.0%, 31/86), fetal distress (24.4%, 21/86), pulmonary infection (23.3%, 20/86), disseminated intravascular coagulation (16.3%, 14/86), multiple organ dysfunction syndrome (16.3%, 14/86), hepatic encephalopathy (9.3%, 8/86), and intrauterine fetal death (2.3%, 2/86). ② Treatment and outcome of AFLP pregnant women: the intensive care unit transfer rate of AFLP pregnant women was 66.3% (57/86). 82 cases were improved and discharged after treatment, 2 cases were transferred to other hospitals for follow-up treatment, and 2 cases (2.3%, 2/86) died. ③ Neonatal outcomes: except for 2 cases of intrauterine death, a total of 106 neonates were delivered, including 39 cases (36.8%, 39/106) of neonatal asphyxia, 63 cases (59.4%, 63/106) of neonatal intensive care unit admission, and 3 cases (2.8%, 3/106) of neonatal death. Conclusions: AFLP is a severe obstetric complication, which is harmful to mother and fetus. In the process of clinical diagnosis and treatment, attention should be paid to the clinical manifestations and laboratory tests of pregnant women, early diagnosis and active treatment, so as to improve maternal and fetal outcomes.
		                        		
		                        		
		                        		
		                        			Pregnancy
		                        			;
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Premature Birth/epidemiology*
		                        			;
		                        		
		                        			Pregnancy Complications/diagnosis*
		                        			;
		                        		
		                        			Fatty Liver/diagnosis*
		                        			;
		                        		
		                        			Fetal Death
		                        			;
		                        		
		                        			Stillbirth
		                        			
		                        		
		                        	
5.Analysis of Differential Proteins Related to Platelet Activation in Patients with Essential Thrombocythemia Based on Label-Free Quantitative Technology.
Yu-Jin LI ; Ju-Ning MA ; Zi-Qin WANG ; Er-Peng YANG ; Ming-Jing WANG ; Jing MING ; De-Hao WANG ; Ji-Cong NIU ; Wei-Yi LIU ; Xiao-Mei HU
Journal of Experimental Hematology 2022;30(3):836-843
		                        		
		                        			OBJECTIVE:
		                        			To analysis the specific protein markers of essential thrombocythemia (ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation.
		                        		
		                        			METHODS:
		                        			Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P<0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P<0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring (PRM) technology.
		                        		
		                        			RESULTS:
		                        			A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing.
		                        		
		                        			CONCLUSION
		                        			Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV, GPVI, COL1A1 and GP1bα can be used as new targets related to ET platelet activation.
		                        		
		                        		
		                        		
		                        			Blood Platelets/metabolism*
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Platelet Activation
		                        			;
		                        		
		                        			Platelet Membrane Glycoproteins/metabolism*
		                        			;
		                        		
		                        			Technology
		                        			;
		                        		
		                        			Thrombocythemia, Essential
		                        			
		                        		
		                        	
6.The Clinical Observation with Ruxolitinib as Graft-Versus-Host Disease Prophylaxis for Children with Thalassemia after Unrelated or Haploidentical Allo-Hematopoietic Stem Cell Transplantation.
Ya-Mei CHEN ; Xiu-Li HONG ; Jin-Zong LIN ; Jie SHI ; Quan-Yi LU
Journal of Experimental Hematology 2022;30(5):1586-1589
		                        		
		                        			OBJECTIVE:
		                        			To retrospectively analyze the efficacy and safety of ruxolitinib therapy for children with thalassemia after unrelated or haploidentical stem cell transplantation.
		                        		
		                        			METHODS:
		                        			From March 2020 to March 2021, 22 patients received successfully allogeneic hematopoietic stem cell transplantation in the Zhongshan Hospital of Xiamen University, from +30 to 100 days,those patients received ruxolitinib therapy (2.5 mg, twice daily) and all adverse reactions were observed, include aGVHD, cGVHD, CMV and EBV infection.
		                        		
		                        			RESULTS:
		                        			22 patients underwent allogeneic stem cell transplantation, 5 patients were diagnosed as aGVHD, 3 patients had grade I-II skin GVHD and 2 patients had grade II intestinal GVHD, those patients were cured. All patients were followed up for more than 21 weeks, 4 cases developed cGVHD, including 3 cases of localized liver GVHD and 1 case of pulmonary GVHD, those were relieved after active treatment. 8 patients had elevated EBV copies (>3×103/ml), and 3 patients had increased CMV copies, the patients recovered after immunosuppressant and antiviral treatment. There was no CMV infection and EBV related post-transplantant lymphoproliferative disorders(PTLD), and no transplant related deaths.
		                        		
		                        			CONCLUSION
		                        			Ruxolitinib can effectively reduce the incidence and severity of GVHD without affecting the hematopoietic recovery, and improve the survival status of thalassemia children after transplantation.
		                        		
		                        		
		                        		
		                        			Antiviral Agents/therapeutic use*
		                        			;
		                        		
		                        			Child
		                        			;
		                        		
		                        			Graft vs Host Disease/prevention & control*
		                        			;
		                        		
		                        			Hematopoietic Stem Cell Transplantation/adverse effects*
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Immunosuppressive Agents/therapeutic use*
		                        			;
		                        		
		                        			Nitriles
		                        			;
		                        		
		                        			Pyrazoles
		                        			;
		                        		
		                        			Pyrimidines
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Thalassemia
		                        			
		                        		
		                        	
7.The Expression and Function of NK Cells in Patients with Acute Myeloid Leukemia.
Lu LIU ; Xiao CHEN ; Hui-Min JIN ; Si-Shu ZHAO ; Yi ZHU ; Si-Xuan QIAN ; Yu-Jie WU
Journal of Experimental Hematology 2022;30(1):49-55
		                        		
		                        			OBJECTIVE:
		                        			To explore the expression characteristics of antigens and functional markers of natural killer (NK) cells in patients with acute myeloid leukemia (AML).
		                        		
		                        			METHODS:
		                        			Multi-parameter flow cytometry was used to detect NK cell surface markers and their functional indicators in 56 newly diagnosed AML patients and 24 healthy controls, including activating receptors NKG2D, NKP46, DNAM-1, and killing indicators granzyme B, perforin.
		                        		
		                        			RESULTS:
		                        			Referring to the WHO hematopoiesis and lymph tissue tumor classification criteria, 56 cases were roughly divided into three types: AML M1, M2, and M4/M5. However, there was no differences about NK cells among the three types, so it was no longer subdivided. NK cells were divided into two groups: CD3-CD56hiCD16- (CD56hiNK) and CD3-CD56dimCD16+ (CD56dimNK). Compared with CD56dimNK cell population, except for NKP46, the positive expression levels of NKG2D and other receptors of CD56hiNK cells in AML patients decreased (P<0.001). Compared with healthy controls, the proportion of CD56hiNK cells in AML patients increased, while the number and proportion of NK cells and proportion of CD56dimNK cells significantly decreased (P<0.05). The proportion of perforin in CD56hiNK cells significantly increased (P<0.05). The expression of DNAM-1 in CD56hiNK cells, NKG2D, DNAM-1, and perforin in CD56dimNK cells decreased significantly (P<0.05). There was no statistically significant difference in expression of other functional indexes in AML patients compared with corresponding indexes of healthy controls. In addition, the proportion of CD56hiNK cells was positively correlated with the expression of CD34+ in AML (r=0.303).
		                        		
		                        			CONCLUSION
		                        			Compared with CD56dimNK, the ratio of CD56hiNK and the expression of functional markers in AML patients are lower. Compared with healthy controls, the number and expression ratio of NK cells in AML patients decrease and the expression of functional markers is abnormal, indicating that its function is impaired.
		                        		
		                        		
		                        		
		                        			CD56 Antigen
		                        			;
		                        		
		                        			Flow Cytometry
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Killer Cells, Natural
		                        			;
		                        		
		                        			Leukemia, Myeloid, Acute
		                        			
		                        		
		                        	
8.Establishment of An Animal Model of Acute B Lymphoblastic Leukemia with Minimal Residual Disease.
Xiao-Fu JIN ; Ling-Zhen WANG ; Ke LEI ; Zeng-Yi DONG ; Ren ZHONG ; Han ZHAO ; Li-Rong SUN
Journal of Experimental Hematology 2022;30(2):381-385
		                        		
		                        			OBJECTIVE:
		                        			To establish an animal model of acute B lymphoblastic leukemia (B-ALL) with minimal residual disease.
		                        		
		                        			METHODS:
		                        			The transplanted tumor was formed by subcutaneous injection of 2×107 Nalm-6 cells, and the body weight, activity status and tumor formation status of nude mice were observed. Peripheral blood, bone marrow, liver and spleen and other tissues of nude mice were taken for pathological examination to understand whether the success of subcutaneous modeling was accompanied by systemic metastasis.
		                        		
		                        			RESULTS:
		                        			There were 2×107 Nalm-6 cells injected subcutaneously in nude mice, (11.0±2.5) days later, the tumors of (3-4) × (3-4) mm were observed, the body weight of the nude mice was reduced and activity showed no limited. Infiltration of tumor cells in liver, spleen and bone marrow were observed in pathological sections.
		                        		
		                        			CONCLUSION
		                        			The animal model of subcutaneous tumor of B-ALL was successfully established in nude mice.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Body Weight
		                        			;
		                        		
		                        			Disease Models, Animal
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Mice, Nude
		                        			;
		                        		
		                        			Neoplasm Transplantation
		                        			;
		                        		
		                        			Neoplasm, Residual
		                        			;
		                        		
		                        			Precursor Cell Lymphoblastic Leukemia-Lymphoma
		                        			
		                        		
		                        	
9.Analysis of DNA Methylation Gene Mutations and Clinical Features in Patients with Myeloproliferative Neoplasm.
Zi-Qing WANG ; Yu-Jin LI ; De-Hao WANG ; Er-Peng YANG ; Yu-Meng LI ; Ji-Cong NIU ; Ming-Qian SUN ; Zhuo CHEN ; Wei-Yi LIU ; Xiao-Mei HU
Journal of Experimental Hematology 2022;30(2):522-528
10.The Clinical Efficacy of Haploidentical Hematopoietic Stem Cell Transplantation by Using Parental Donors in Patients with Thalassemia.
Han-Bing OU ; Jin-Zong LIN ; Xiu-Li HONG ; Jing-Yuan LU ; Quan-Yi LU
Journal of Experimental Hematology 2022;30(2):534-538
		                        		
		                        			OBJECTIVE:
		                        			To analyze the clinical efficacy of haploidentical hematopoietic stem cell transplantation (haplo-HSCT) by using parental donors on thalassemia patients.
		                        		
		                        			METHODS:
		                        			The 13 thalassemia patients treated by haplo-HSCT using parental donors in our hospital from July 1, 2016, to July 1, 2020 were retrospectively reviewed. Hematopoiesis reconstitution, the incidence of GVHD, infections and the long-term survival of the patients were analyzed.
		                        		
		                        			RESULTS:
		                        			Twelve of the 13 patients were successfully implanted, the success rate of implantation was 92.3%. The median time of neutrophil and platelet engraftment was 12.5 days (range, 9-22 days) and 21 days (range,12-34 days), respectively. One patient achieved primary graft failure. Three (25%) patients developed to acute GVHD (aGVHD) and achieved complete remission after treatment. Chronic GVHD developed in three (25%) patients, one of them was extensive and under treatment, while one patient developed to severe bacterial infection (7.7%). CMV viremia was diagnosed in two patients (15.4%). There were no patients developed to CMV disease. Three (23.1%) patients achieved EB viremia after transplantation, one of them developed to EBV-related lymphocytic proliferative disease, while there were no patients showed invasive fungal infection. At the last follow-up, all patients survived, twelve of them were free from transfusion dependency. There were no transplant-related deaths. Projected overall and thalassemia-free survival at three years was 100% and 92.3%, respectively.
		                        		
		                        			CONCLUSION
		                        			The transplant protocol of haplo-HSCT by using parental donors in patients with thalassemia has reliable source of donors, high incidence of successful implantation and low incidence of GVHD, which can be used as an effective way to increase the source of donors in children with thalassemia.
		                        		
		                        		
		                        		
		                        			Child
		                        			;
		                        		
		                        			Cytomegalovirus Infections
		                        			;
		                        		
		                        			Graft vs Host Disease
		                        			;
		                        		
		                        			Hematopoietic Stem Cell Transplantation
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Parents
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Thalassemia/therapy*
		                        			;
		                        		
		                        			Transplantation Conditioning/methods*
		                        			;
		                        		
		                        			Treatment Outcome
		                        			;
		                        		
		                        			Viremia
		                        			
		                        		
		                        	
            
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