Objective: To summarize the laboratory findings of a case of hemolytic disease of the fetus and newborn (HDFN) caused by Rh system anti-c antibodies and to review the literature, so as to explore the characteristics of anti-c HDFN. Methods: The ABO blood type, Rh blood type, direct antiglobulin test (DAT) results, and the presence of unexpected antibodies and their titers were determined by serological methods. The cases of anti-c HDFN in our laboratory in China and abroad were statistically analyzed, and the incidence of severe HDFN caused by anti-c, anti-D and anti-E was compared. Results: The blood type of the child was B (Rh CcDee) with a positive DAT. Anti-c antibody was detected in both serum and eluate, with a serum antibody titer of 4. The mother’s blood type was AB (Rh CCDee) with a negative DAT, and anti-c antibody was detected in the serum with a titer of 128. Among 20 cases of anti-c HDFN, 17 were DAT positive, and 9 (45%, 9/20) underwent blood transfusion or exchange transfusion. The incidence of severe HDFN was 47.60% (10/21) for anti-c, 47.60% (10/21) for anti-D and 31.30% (5/16) for anti-E. Conclusion: Maternal pregnancy and/or blood transfusion are the main reasons for the production of Rh alloantibodies such as anti-c. The prevention and management of anti-c should be similar to that of anti-D. Rh antigen-matched (five antigens of Rh blood group) transfusion is necessary for women of childbearing age to avoid antibody production, and Rh typing and antibody screening during prenatal examination is recommended to ensure early detection, intervention and treatment.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Messenger RNA (mRNA) can be modified by more than 100 chemical modifications. Among these modifications, N6-methyladenosine (m⁶A) is one of the most prevalent modifications. During the processes of cells differentiation, embryo development or stress, m⁶A can be modified on key mRNAs and regulate the progress of cells through modulating mRNA metabolism and translation. Other mRNA modifications, including N1-methyladenosine (m¹A), 5-methylcytosine (m⁵C) and pseudouridine, together with m⁶A form the epitranscriptome of mRNA that accurately modulate the mRNA translation. Here we review the types and characteristic of mRNA epigenetic modifications, especially the recent progresses of the function of m⁶A, we also expect the main research direction of m⁶A epigenetic modification in the future.
Adenosine ; analogs & derivatives ; genetics ; metabolism ; Cell Differentiation ; genetics ; Embryonic Development ; genetics ; Epigenesis, Genetic ; Gene Expression Regulation ; RNA Processing, Post-Transcriptional ; RNA, Messenger ; metabolism

To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals ; Baculoviridae ; genetics ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Immune Sera ; Insecta ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Retinol-Binding Proteins, Plasma ; biosynthesis ; immunology ; Sf9 Cells ; metabolism ; Transfection

Objective To determine the effects of trimetazidine (TMZ) on pathology and energy metabolism of myocardium in chronic renal failure(CRF) rats.Methods CRF models were built in Sprague-Dawley (SD) rats with 5/6 subtotal nephrectomy, and animals were randomyly divided into sham group, control group and three groups treated with different doses of TMZ (3 mg/kg,6 mg/kg or 9 mg/kg).TMZ was intragastrically administrated to CRF rats for 17 weeks, while physiologicalsalinewasusedascontrol. Transthoracicechocardiographywasperformedand myocardial morphosis was observed.Left ventricular weight/body weight(LVW/BW) and heart weight/body weight (HW/BW) were measured, and heart rate, and mean arterial pressure (MAP)were detected at the end of the study, while several parameters were detected, including urea nitrogen (BUN), creatinine(Scr), triphosaden(ATP), adenosine diphosphate(ADP), superoxide dismutase (SOD), malondialdehyde (MDA), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α).Results (l)Left ventricle end-systolic dimensions, anterior wall end-diastolic and end-systolic thicknesses, and posterior wall end-diastolic thickness were significantly lower in rats treated with either medium dose or high dose of TMZ, as compared with control group(P＜0.05).(2)LVW/BW and HW/BW in rats treated with either medium dose or high dose of TMZ were significantly lower than those in control group(P＜0.05). (3)Various pathological changes were observed in control group, such as irregular arrangement and hypertrophy of the cardiomyocytes, myocardial fibrosis,mitochondrial swelling, focal muscle fiber dissolution, etc.However, all these pathological changes were apparently ameliorated in TMZ-treated groups, while the beneficial effects of TMZ therapy were dose-dependent. (4)No difference was observed in heart rate among all the groups.Although no difference existed in all the CRF rats, concerning on the systolic/diastolic blood pressure and mean arterial pressure (P＞0.05), these parameters were elevated in CRF rats, as compared with sham-operated group(P＜0.01). (5)ATP and ADP in TMZ-treated rats were significantly higher as compared with control(P＜0.05), moreover, medium dose and high dose of TMZ were superior to low dose (P＜0.05).(6)SOD was significantly increased in TMZ-treated rats (P＜0.05), while IL-6,TNF-α and MDA were significantly decreased in medium dose and high dose of TMZ, as compared with control(P＜0.05).Conclusion TMZ may prevent myocardial fibrosis and left ventricular hypertrophy in chronic renal failure via ameliorating myocardial energy metabolism and alleviating inflammatory reaction and oxidative stress.

Objective To investigate the effects of IL-12 coexpression level on antigen expression and immune responses induced by HBsAg DNA vaccination. Methods DNA vaccine plasmid pHBV carrying codon-optimized preS2-S gene of reference sequence CHN-HBV07-C in China was constructed. Three DNA vaccine plasmids pHBV-12i, pHBV-12l and pHBV-12h were also constructed by subcloning three different IL-12 expression cassettes with various expression strengths to plasmid pHBV, respectively. Expression levels of IL-12 and HBsAg in vaccine plasmid-tranfected 293T cells were measured by quantitative ELISA. DNA vaccines were administered intramuscularly to BALB/c mice and HBsAg-specific cellular immune responses were determined by IFN-γ ELISPOT. HBsAg-specific antibodies were tested by Chemiluminescence Quantitative Immunoassay. Results The HBsAg expression level in 293T cells was 70 ng/ml when transfected by plasmid pHBV without IL-12 expression cassette, and the HBsAg level was 18 ng/ml when transfected by plasmid pHBV-12l carrying low-level IL-12 expression cassette, whereas the HBsAg level was only 6 ng/ml when transfected by plasmid pHBV-12h carrying high-level IL-12 expression cassette.Results of DNA vaccination revealed that HBsAg-specific humoral and cellular immune responses were significantly decreased in mice administering vaccine pHBV-12h carrying high-level IL-12 expression cassette. Although HBsAg-specific antibody responses in mice inoculated with pHBV-12l were also decreased when compared with those in pHBV-vaccinated mice without IL-12 expression, the HBsAg-specific cellular immune responses were significantly increased. Conclusion High-level coexpression of IL-12 may suppress the expression of HBsAg, Whereas modest coexpression of IL-12 significantly enhanced the HBsAg-specific T cell responses induced by DNA vaccination. Therefore, it is so important to balance the expression between adjuvant and antigen to enhance the immune response.

Objective:Clonidine,by activating peripheral α-sbrenoceptors, produces transient pressor response after i.v.injection in anesthetized animals.Moxonidine, with at least 40-fold higher affinity to I1-imidazoline receptors than to α2-adrenoceptors,produces also a transient pressor response. This work was designed to investigate whether I1-imidazoline receptors are involved in this pressor effect of moxonidine. Methods:Female spontaneously hypertensive rats(SHRs,aged 14-16 weeks)were anesthetized with urethane.To observe the transient pressor responses,moxonidine 0.1,0.3,1.0mg/kg(intravenous,i.v),2.0μg(intracerebroventricular,i.c.v.)and 1.0,10.0mg/kg(intragastric,i.g.)were administrated in different groups of rats.To evaluate the roles of α1-adrenoceptors,α2-adrenoceptors and I1-imidazoline receptors in the transient pressor responses to moxonidine, prazosin(10.0μg/kg),yohimbine(2.0mg/kg),phentolamine(0.2mg/kg),idazoxan(1.0mg/kg)or yohimbine+idazoxan(2.0mg/kg+1.0mg/kg)were intravenously given to the animals before moxonidine 0.3mg/kg (i.v.).Results:It was found that i.v.moxonidine produced a greater pressor response than clonidine when producing a similar reduction of blood pressure.This effect of moxonidine was not influenced by prazosin, but was partly inhibited by yohimbine, phentolamine or idazoxan,and completely blocked by the combination of yohimbine and idzaxon.Neither i.c.v.injection nor i.g. administration of moxonidine induced transient pressor responses.Conclusion:The transient pressor response of i.v. moxonidine is mediated by both peripheral I1-imidazoline receptors and α2-adrenoceptors.

Scaffolds provide a three-dimensional environment that is desirable for inducing and promoting the production of cartilaginous tissue.Ideally the scaffold should:① have directed and controlled degradation;② promote cell viability,differentiation,and extracellular matrix production;③ adhere and integrate with the surrounding native cartilage;④ span and assume the size of the defect;⑤ provide mechanical integrity depending on the defect location;⑥ have no toxicity,no immunogenicity or inflammatory induction;⑦ allow for the diffusion of nutrients and waste products.To date,a wide range of natural and synthetic materials have been investigated as a scaffold for cartilage repair.Based on the morphology and structure,these materials can be divided into hydrogel,sponge,fiber mesh and so on.Natural polymers that have been explored as bioactive scaffolds for cartilage engineering include:alginate,agarose,fibrin,hyaluronic acid,collagen,gelatin,chitosan,chondroitin sulfate,and cellulose.Synthetic polymers currently explored for cartilage repair include poly(?-hydroxy esters),polyethylene glycol,poly(NiPAAm),poly(propylene fumarates),and polyurethanes.

Stimulating factors have been employed to induce,accelerate,and/or enhance cartilage formation.For instance,stimulating factors and other additives may be added to culture media in vitro or incorporated into scaffolds for in vivo delivery to control cellular differentiation and tissue formation.At present,many growth factors and other dissoluble factors such as hyaluronic acid,chondroitin sulfate,and insulin have been used alone or synergistically in cartilage tissue engineering.The application efficacy depends on cell type and culture condition.In addition,gene therapy has emerged as another method of local delivery,where cells can be engineered to over-express bioactive molecules.An additional approach is the introduction of mechanical signals through loading regimes such as hydrostatic or dynamic compression or through the use of bioreactors.Current bioreactors used in cartilage tissue engineering include flat-sheet bioreactor,rotating wall vessel bioreactor and concentric cylinder bioreactor.Bioreactor can improve nutrition transmittability,provide hydrodynamic environment,induce shear stress,and promote production of cartilage specific matrix protein.

Objective:Glucose-insulin-potassium(GIK) is clinically used for reducing mortality in acute myocardial infarction(MI). It is known that ventricular arrhythmia, left ventricular dysfunction and impaired baroreflex sensitivity(BRS) are the three major determinants for predicting the mortality after acute MI. The present work was designed to study the effects of GIK on BRS, ventricular arrhythmia, and left ventricular function in rats with coronary artery ligature. Sprague-Dawley rats were used and the myocardial infarction was produced by ligature of the left anterior descending artery. Five weeks after coronary artery ligation, BRS was measured in conscious state with a computerized blood pressure monitoring system and left ventricular function and electrocardiogram were determined in the anaesthetized state in the subacute phase of myocardial infarction. It was found that GIK did not affect the blood pressure and heart period in both conscious and anaesthetized rats. GIK did not enhance BRS, but reduced ventricular arrhythmia and improved left ventricular function by reducing left ventricular end diastolic pressure in anaesthetized rats with MI. It is proposed that reducing ventricular arrhythmia and improving left ventricular function contribute to the effect of GIK on reducing the mortality after MI.