Objective To investigate the efficacy of HLA-matched ABO-incompatible sibling al- logeneic peripheral blood stern cell transplantation(allo-PBSCT)in treatment of hematologic malig- nancies.Methods Between June 2001 and September 2005,68 patients with hematologic malignancies received allo-PBSCT from HLA-identical sibling donors.Among them,38 were ABO-compatible (ABO-compatible group),30 ABO-incompatible(ABO-incompatible group).Conditioning regimen of all patients:BU/CY used as conditioning regimen in AML,MDS and CML patients;TBI/CY used in ALL and NHL patients.TBI/CY/Melphan used in MM patients.GVHD prophylaxis:A combination of cyclosporine(CsA)and mycophenolate mofetil(MMF)and methotrexate(MTX)was administered for GVHD prophylaxis.Results (1)Sixty-seven patients were all engrafted,and only one patient was graft failure.The median time(range)to neutrophils≥0.5?10~9/L and platelets≥20?10~9/L was +12(+9～+15)and+21(+15～+40)days post-transplant respectively.The time to neutrophil and platelet engraftment had no significant difference between ABO-incompatible group and ABO-com- patible group(P＞0.05);(2)Thirty patients with ABO-incompatible allo-PBSCT had no evidence of hemolysis.In ABO-incompatible group,the onset of erythropoiesis after allo-PBSCT was delayed. Three of 7 patients with blood group“O”receiving a group“A”developed pure red cell aplasia (PRCA).The blood type of 30 patients would transfer to the type of donors in 60 days(24～153 days)after transplantation.(3)Following up to 30th,Sep.2005,the incidence of aGVHD was high- er in ABO-incompatible group(20.0%)than that in ABO-compatible group(2.6%,P=0.019). The incidence of cGVHD,VOD,HC,CMV infection,disease relapsed rates and mortality had no sig- nificant difference between ABO-incompatible and ABO-compatible recipients(P＞0.05).(4)Kaplan- Meier survival analysis revealed the rates of survival had no significant difference between ABO-incom- patible and ABO-compatible recipients(P＞0.05).Conclusions ABO-incompatible allo-PBSCT is fairly safe if there is indication.Although the incidence of aGVHD was higher in ABO-incompatible group than that in ABO-compatible group,but there was no effect on engraftment,incidence of cGVHD or prognosis.

Objective:To summarize the oblique branches found in the transfer of anterolateral thigh flap (ALTF) and to handle the issues of oblique branch.Methods:Thirty patients who require surgery of ALTF transfer from May, 2017 to October, 2019 were enrolled. CTA examination was perfected prior surgery and the origin of ALTF vessels was preliminarily determined. During the surgery, Three-longitudinal-and-five-transverse methods were used to locate and design the flap. The ALTF was taken according to the flap design. Attention should be paid to the location of the oblique branch and the separation of the vessels of oblique branch to avoid a damage as much as possible. The vessels of oblique branch should be completely explored and separated. One or 2 vascular pedicles were cut according to whether the oblique branch vessels and the descending branch vessels were joined together during surgery. Clamping tests were carried out on the 2 vascular pedicles to determine an arterial blood supply. Super drainage of vascular pedicle veins according to the situation of blood circulation. After the surgery, routine treatment was carried out. Blood supply, skin temperature, swelling degree, exudation and survival of the flap were closely observed and regular follow-up was carried out.Results:Among the 30 ALTF examined by CTA, 13 patients were identified with oblique branch vessels before operation. During operation, 11 oblique branch vessels (The occurrencce rate was 36.6%)were found to enter the flap, and were completely preserved. Of the 11 identified oblique branch vessels, 8 had 2 vascular pedicles taken and the vascular pedicles were treated by venous super drainage technique. The postoperative blood supply of the flap was good; The skin temperature was closed to surrounding normal skin; Swelling of flap was minor and there was little seepage. The flaps all survived after surgery with stage one healing. Followed-up time was 3-32 (average 16.1) months. The recipient site healing was good, and the function and appearance were satisfactory. The joint movement at the donor site was normal, and there was no obvious loss of local sensation.Conclusion:More than one third of the oblique branches appear in this group. The oblique branch vessels should be preserved as much as possible to avoid issues in relation to the oblique branch. Reasonable handling of oblique branch is the key to the success of the surgery.

Objective To explore the essence of polycystic ovary syndrome of spleen-kidney yang deficiency,and to investigate the objective standardization of TCM syndrome diagnosis of the disease.Methods Among the 200 PCOS patients,the Chinese medicine syndrome pattern was differentiated as spleen-kidney yangdeficiency type (n=105 )and Non-spleen-kidney yang deficiency type (n=95).To measure and compare their levels ofBMI、WHR、sexual hormones、FINS、2 hINS,and HOMR-IR.Results The levels of BMI (24.4±2.8)、WHR (0.88±0.07)、LH/FSH (2.02±1.06)、E2 (55.84±23.87) pg/ml、HOMR-IR (2.95±1.52)、FINS (15.18±7.64) IU/ml、2hINS ( 59.53 ±40.61 ) IU/ml in the group of spleen-kidney yang deficiency type were obviously higher than those[ (22.2±2.2)、(0.80±0.06)、(3.50± 1.80)、(45.82±21.31) pg/ml、( 1.90± 1.50)、(9.93±6.59) IU/ml、(50.56±41.34) IU/ml respectively] in the group of Non-spleen-kidney yang deficiency type (P＜0.05＝,but T and FSH level showed no significant difference between two groups [FSH the group of spleen-kidney yang deficiency type=(6.46±0.95) IU/L、FSH the group of No-spleen-kidney yang deficiency type=(6.40±1.16) IU/L; T the group of spleen-kidney yang deficiency type=(0.81±0.15) μg/L、T the group of No-spleen-kidney yang deficiency type=(0.72±0.13) μg/L,P＞0.05].Conclusion BMI、WHR、LH/FSH、E2、FINS、2HINS,and HOMR-IR could be used as objective index in identifying the difference in between spleen-kidney yang deficiency type and Non-spleen-kidney yang deficiency type.

Objective To establish and identify the induced pluripotent stem cell(iPSC) line reprogrammed from human umbilical cord mesenchymal cells(HuMSCs).Methods HuMSCs were cultured by adhesion method,and OCT4,SOX2,KLF4,c-Myc,NANOG,LIN-28 were transfected into HuMSCs with lentiviral victor to reprogramme HuMSCs into iPSC.Morphological observation,pluripotency genes (SOX2,TDGF1,THY-1,OCT4,REX1 and TERF1) expression,alkaline phosphatase detection,karyotype analysis,embryonic stem cells (ESC) specific proteins (NANOG,OCT4,SSEA-4,TRA-1-81) immunofluorescence staining,differentiated into teratomas in vivo(inject the iPSC into SCID mice) and embryniod bodies in vitro were performed to exam the pluripotency of the iPSC.Results Four days after being infected by lentivirus,the HuMSCs became round-shape; 10 days after infection,some embryonic stem(ESC)-like colonies appeared.Fourteen days after infection,picked up the regularly shaped colonies and cultured several passages.About 1.25％ HuMSCs were reprogrammed into iPSC.The iPSC presented clone-like growth like ESC.All the cells were positive to alkaline phosphatase staining and expressed the pluripotency genes.The iPSC also expressed the ESC specific proteins,and karyotype analysis showed normal chromosome caryotype (46,XY).Furthermore,the iPSC could form embryoid bodies in vitro,expressed alpha fetoprotein(AFP),smooth muscle actin(SMA) and β-tubulin.The iPSC could alsoform teratomas in vivo.Conclusion OCT4,SOX2,KLF4,c-Myc,NANOG,LIN-28 can reprogram HuMSCs into iPSC efficiently.

OBJECTIVETo study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.

METHODSPorcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).

RESULTSTen pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).

CONCLUSIONEffective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.

Androgens ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flavonoids ; Granulosa Cells ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinases ; metabolism ; RNA, Messenger ; Swine

Objective To investigate the extracellular polysaccharide distribution and components of Ureaplasma urealyticum (Uu) after biofilm having been developed in.Methods The standard serotype 3 and serotype 14 belong to biovar Parvo,and the standard serotype 4 and serotype 8 belong to biovar T960 were employed to form biofilrns in vitro.Scanning electron microscope and confocal laser scanning microscope were used to analysis the biofilms and extracellular polysaccharide.We used combination of two different labeled lectins,Canavalia ensiformis(FITC-ConA) and Erythrina cristagalli(ECA) which bind to specific polysaccharide residues to visualize extracellular polysaccharide in biofilms,and average uorescence intensity was evaluated Results All the strains can form the biofilmsin vitro.The biofilm was honeycomb-Like structures mainly,and extracellular polymeric substances accounts for majority of proportions.All the extracellular polysaccharide could be combined with FITC-ConA and ECA,and the total average fluorescence intensity of FITC-ConA was higher than ECA( P＜0.001 ).Conclusion Ureaplasma urealyticum biofilm is honeycomb-like structures mainly.The extracellular polysaccharide contains,galactose,and N-acetyl glucan residual,and the glucose,mannose residual are the main components.

Objective:To recognize the characteristics of necrotizing fasciitis patients complicated with sepsis and summarize the experience the treatment.Methods:A retrospective study was conducted. The clinical data of 57 patients with necrotizing fasciitis complicated with sepsis admitted to Guangdong Provincial People's Hospital from July 2009 to December 2019 was analyzed by collecting such factors as gender, age, complications, infection sites, pathogens, surgery information, treatment options and outcome. The patients were divided into debridement group ( n = 14) and control group ( n = 43) according to whether the debridement was completed within 48 hours of admission, and the mortality during hospitalization between the two groups was compared. A telephone follow-up had been done to record the long-term outcome of these patients. Results:Among 57 patients with necrotizing fasciitis complicated with sepsis, there were 43 males and 14 females with the average age of (57.9±12.1) years old. Most of the underlying diseases were diabetes mellitus (70.17%), other diseases included hypertension (8.77%), tumor chemotherapy (7.02%), liver disease (hepatitis, cirrhosis, 7.02%), coronary artery heart disease (3.51%), systemic lupus erythematosus (3.51%), etc. Most of the infection site was lower limbs (71.93%). There were 78 pathogens cultured in 57 patients, in which 52 were non-drug resistant bacteria (66.67%), and 26 were drug resistant bacteria (33.33%). There were 40 Gram positive (G +) bacteria (51.28%), 29 Gram negative (G -) bacteria (37.18%), 8 fungi (10.26%) and 1 mixed bacteria (1.28%). Finally, of 57 patients, 46 patients were cured, and 11 patients died with hospital mortality of 19.30%. Among 57 patients, the hospital mortality in the debridement group was significantly lower than that in the control group [0% (0/14) vs. 25.58% (11/43), P < 0.05]. Among the 46 cured patients, 11 had accepted amputations, accounting for 23.91%. In December 2020, 43 patients who were cured (3 patients were lost to follow-up) were followed up by telephone. Twenty-three patients were completely self-care, 9 patients were partly self-care, 8 patients were completely unable to take care of themselves, and 3 patients died. Conclusions:Necrotizing fasciitis with sepsis mostly occurs in people with weakened immunity, and has a high mortality and disability rate. Early identification and active surgical debridement may be the key to improve the treatment effect.

Rosai-Dorfman disease is a rare, idiopathic, benign, and self-limiting histiocytic proliferative disorder. A 26-year-old man presented with a single massive cutaneous nodule (reaching 30 cm in diameter) on the left shoulder and back for 15 months. The routine hematological and biochemical tests were normal. Magnetic resonance scanning showed the lesion involved the skin, subcutaneous tissue, and subjacent muscle group, accompanied by obvious lymph node enlargement in the left part of the neck, supraclavicular fossa, and axillary fossa. The histopathology of the left cervical lymph node revealed diffuse effacement of the normal nodal architecture, with patchy chronic inflammatory cell infiltrates comprising lymphocytes and sheets of histiocytes. Some histiocytes contained lymphocytes within their pale cytoplasm. Many multinucleated giant cells were found; however, caseating granulomas were not seen. The skin and muscle biopsy specimen obtained from the back revealed infiltrating lymphocytes and histiocytes diffusely distributed in the dermis, subcutaneous tissue, and crevices of the muscle fibers. The phenomenon of emperipolesis and the presence of multinucleated giant cells were also seen. Immunohistochemical staining revealed that the histiocytes were positive for S-100 protein and CD68 but negative for CD1a. Immunophenotyping of the infiltrating lymphocytes indicated positive reactions to CD3, CD45RO, CD5, CD7, CD4, CD8 (partly), CD79a, CD20 (partly), and Ki-67 (<1%). The final diagnosis was Rosai-Dorfman disease. Owing to the extensive and deep involvement of the subcutaneous tissue and muscles, the patient did not undergo surgery to excise the massive skin nodule. The lesion showed no obvious change at the 12-month follow-up.
Adult ; Biopsy ; Cytoplasm ; Dermis ; Diagnosis ; Emperipolesis ; Follow-Up Studies ; Giant Cells ; Granuloma ; Histiocytes ; Histiocytosis, Sinus* ; Humans ; Immunophenotyping ; Lymph Nodes ; Lymphocytes ; Muscles ; Neck ; S100 Proteins ; Shoulder* ; Skin ; Subcutaneous Tissue

OBJECTIVETo investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro.

METHODSThree fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation.

RESULTSThree recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC.

CONCLUSIONSFunctional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.

Antigens, CD ; pharmacology ; B7-2 Antigen ; Escherichia coli ; genetics ; Humans ; Immunoglobulin Constant Regions ; pharmacology ; Immunoglobulin Variable Region ; pharmacology ; Lymphocyte Activation ; Membrane Glycoproteins ; pharmacology ; Plasmids ; Recombinant Fusion Proteins ; pharmacology ; T-Lymphocytes ; immunology

Female ; HIV Infections ; prevention & control ; transmission ; HIV-1 ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Pregnancy ; Risk Factors