[Abstract] [Objective] To further identify the RhD phenotype and RHD genotype in the individual who have RhD negative phenotype in the primary screening, and to analyze the effect of c. 801+2T>G mutation on RhD phenotype by minigene splicing assay. [Methods] The serologic test was performed for RhD phenotype identification and absorption-elution test was performed by using monoclonal anti-D. Sanger sequencing was used to analyze the sequence of RHD genes and the newly identified splicing site mutations of RHD genes were used to construct pSplicePOLR2G micro gene expression plasmids. By using an in vitro micro gene splicing system, the mRNA splicing results were detected and analyzed using agarose and capillary electrophoresis to predict their impact on RhD phenotype. [Results] The serological test results showed that the patient's blood type was RhD-negative, but the anti-D absorption-elution test was positive, indicating a Del phenotype. The rare genotype RHD^*(1227A/801+2G) was identified in this individual. The c. 801+2T>G was a novel mutation at 5'-splice site of intron 5. The minigene splicing assay showed that c. 801+2T>G resulted in a complete skipping of RHD exon 5 in the mature transcript, forming a transcript without exon 5. [Conclusion] An individual carrying a novel mutation c. 801+2T>G in the RHD gene was found to exhibit a Del phenotype, but also carry the Asian Del allele c. 1227G>A. It was speculated that the c. 801+2T>G mutation caused RhD negative or Del phenotype based on the results of minigene splicing assay in vitro.

【Objective】 To identify the specificity of alloantibody against high-frequency antigens in one case suffering with severe hemolytic diseases of the fetus and newborn (HDFN) and to screen for matching blood for transfusion. 【Methods】 The HDFN test and the antibody serological identification tests in the mother were performed. Several common high frequency antigens of maternal red blood cells (RBCs) were determined. IgG subtype coated on the RBCs of the newborn was determined. The phagocytic efficiency of the antibody was tested using the monocyte phagocytosis of sensitized erythrocyte by flow cytometry in vitro. Sanger sequencing of DI gene was performed in the mother, father and mother’s brother. The diluted maternal plasma was used for large scale screening of matching blood using IAT in Coomb’s gel card. 【Results】 Di(b-) phenotype was identified in the mother of the newborn and anti-Di^b (titer: 512) related HDN was detected in the newborn. IgG1 and IgG2 subtypes of anti-Di^b were detected and the rate of monocyte phagocytosis was 88.83%(74.7/84.09). The compatible blood was not detected in the maternal relatives. Subsequently, the newborn received the matching RBCs of two Di(b-) donors identified from 5 520 blood donors and discharged from the hospital. We screened out 17 Di(b-) donors out of 51 334 blood donors, indicating that the distribution frequency of Di(b-) among blood donors in Guangzhou was about 0.033% (17/51 334). 【Conclusion】 By serology and molecular biology methods, the newborn was identified with HDFN caused by anti-Di^b, and an effective large-scale screening method for Di (b -) rare blood types was established to find matching blood, which supported the establishment of rare Di(b-) blood database.

Objective To summarize the current situation of the implementation of the death case discussion system in county-level medical institutions and analyze the existing problems,to attempt to make suggestions for improvement.Methods Judgmental sampling method was used in this study.A self-made questionnaire was used to conduct field investigation on the im-plementation of the death case discussion system in 16 county-level medical institutions in Chongqing,and semi-structured inter-views were conducted with medical staff.Results All county-level medical institutions have established death case discussion system,but there are many differences in connotative quality.There are many problems in system implementation,such as unclear scope of death case discussion,formality,irregular discussion,poor quality of connotation and lack of systematic supervision.Conclusion As a core medical system,the death case discussion system is an important tool for medical quality management.The medical quality management and medical staff must place a high value on the significance of death case discussion.In order to enhance medical quality and ensure patient safety,we must adhere to the principles of openness and honesty,systematic criti-cism,non-recrimination and non-punishment to conduct death case discussion in time and standardize,keep a record of death case discussions and enhance supervision.

【Objective】 To solve the difficulty of RhD blood group typing in a patient with double population(DP) of red blood cells for RhD antigen by serological and genotyping analysis. 【Methods】 Separation of the two populations of red blood cells of the patient was performed using capillary centrifugation method. ABO, RhD and RhCE typing, direct anti-human globulin test (DAT), irregular antibody screening, antibody identification and blood crossmatching of the patient were conducted using the standard serological methods. The hybrid Rhesus zygosity analysis of the RHD gene was performed by PCR-RFLP method. RHD and RHCE genotype of the patients were identified by PCR-SSP method. 【Results】 The patient was B type but with DP of red blood cells for RhD, Rhc and RhE antigens. DAT of the patient was positive and the alloanti-D was detected in serum. The RHD zygosity was D-/D- homozygote. PCR-SSP testing showed the RHD gene deletion (RHD * 01N. 01/01N.01 genotype) and Ccee of RHCE genotype in the patient, which was consistent with RHD zygosity analysis. 【Conclusion】 This is a special case with D-negative phenotype which was wrongly detected as D-positive type after D-positive red blood cells transfusion in emergency. When the DP of red cells for D antigen encountered like this case, the RhD typing can be accurately determined by using RHD genotyping analysis to provide strong evidence to the clinical blood transfusion.

【Objective】 To identify the antibody specificity in a pregnant women who had no history of blood transfusion but presented the antibodies against high-frequency antigens. 【Methods】 ABO, RhD blood group antigens were identified by saline. Antibody screening and identification were performed by saline and indirect Coomb’s technique. Further antibody identification tests were conducted using papain, trypsin and chymotrypsin-treated cells. Antibody titer in serum was tested. PCR amplification and sequencing analysis of 16 exons of ABCG2 gene were conducted. 【Results】 The blood type of the patient were B, RhD positive. The serum reacted with antibody screening/identified cells by indirect antiglobin test(both 2+ ) but not by saline. The agglutination was enhanced after papain treatment (4+ ), but remained unchanged after trypsin and chymotrypsin treatment (2+ ). The IgG titer was 1∶2. The sequencing analysis of ABCG2 gene revealed a homozygous nonsense mutation(c.376C>T, p. Gln126X) in exon 4 of the women. 【Conclusion】 In this case, the development of anti-Jr^a in Jr(a-) mother was stimulated by mother-child serology incompatibility during pregnancy.

【Objective】 To identify the blood group epitope of a D variant individual and analyze its molecular characteristics. 【Methods】 The saline test and indirect antiglobulin test (IAT) were used to identify the RhD serologically. The anti-human globulin gel card was used for direct antiglobulin test (DAT). RhD epitopes were detected using the epitope detection kit (D-Screen). RhCE antigens were typed using Rh typing Card. The RHD gene zygomorphism was further analyzed by PCR-RFLP. Ten exons of RHD gene were amplified by PCR and analyzed by direct sequencing. 【Results】 DAT test was negative, and the serological results showed weak expression of RhD, which was D variant. The RhD epitope test results showed that the red blood cells of this patient had a weak agglutination with 4 monoclonal anti-D against epD6.4, epD6.1, epD2.1, and epD5.4 (w+ to 2+ ), and reacted negatively with other epitope antibodies. RhCE antigen typing was Ccee; The RHD gene zygomorphism result was D+ /D-, the sequencing of RHD exons revealed that the first exon carried c. 41C>T (p.Pro14Leu) missense mutation, and its genotype was RHD^*01W.136/01N.01. 【Conclusion】 This D variant is the first weak D type 136 reported in the Chinese population, and its phenotype is weak partial D.

Objective: To evaluate the anti-gingivitis efficacy of a novel stabilized stannous-containing sodium fluoride dentifrice. Methods: A randomized, controlled and double blind clinical study was conducted. 156 adults with gingivitis were enrolled and randomly assigned to experimental group(group of novel stabilized stannous-containing sodium fluoride dentifrice, n = 51), positive control group (group of Yunnan Baiyao dentifrice, n = 54) and negative control group (group of Crest dentifrice, n = 51). Gingival health was assessed using Mazza Modification of the Papillary Bleeding Index(Mazza GI) at Baseline, day 3 and week 4 and pocket depth was evaluated at baseline and week 4, respectively. Results: At day 3 and week 4, the experimental and positive control groups exhibited lower clinical parameters than the negative control group(P＜ 0. 000 1). At week 4, the mean Mazza GI scores and PD of the experimental group were significantly lower than those of the positive control group(P＜ 0. 05). Conclusion: The novel stannous-containing sodium fluoride dentifrice has anti-gingivitis efficacy.

Objective To analyze the clinical characteristics of IgG4-related disease (IgG4-RD)so as to improve the understanding of IgG4-RD in China.Methods IgG4-RD patients were recruited from Peking Union Medical College Hospital between January 2011 and January 2016.All patients were followedup for more than 6 months.The demographic characteristics,symptoms,organ involvements,laboratory examinations and treatment efficacy were evaluated and analyzed.Results A total of 346 patients were finally enrolled,including 230 males (66.5％) and 116 females (33.5％).The mean age of disease onset was (53.8 ± 14.2) years old.The mostly common involved organs were lymph nodes (56.4％) and submandibular glands (52.6％).Other affected organs and manifestations included:swelling of the lacrimal glands (46.5％),autoimmune pancreatitis (38.4％),pulmonary involvement (28.0％),sclerosing cholangitis (25.4％),naso-sinusitis (23.4％),parotid gland swelling (21.7％),retroperitoneal fibrosis (19.9％),large arteries involvement (9.5％),kidney involvement (obstructive nephropathy caused by retroperitoneal fibrosis was excluded) (6.9％),skin lesions (6.4％).Rare features consisted of thyroid glands,pituitary glands,gastrointestinal tract,pachymeningitis,pericardium,sclerosing mediastinitis and orchitis.The majority of patients had multi-organ involvement,such as 74.3％ patients with 3 and more,18.2％ and 7.5％ patients with 2 and single organ involvement respectively.The average IgG4-RD responder index (IgG4-RD RI) was 13.21 ±5.70.History of allergy was found in 172 (49.7％) patients.As to the laboratory tests,elevated serum IgG4 levels were confirmed in 285 (94.1％) patients,which was positively correlated with IgG4-RD RI.There were 33.5％ patients receiving monotherapy of glucocorticoid,52.6％ treated with glucocorticoids combined with immunosuppressive agents,4.9％ patients with immunosuppressant only,and 9.0％ patients with mild disease not receiving medication.The majority (336,97.1％) patients improved the above regimens.Conclusion IgG4-RD is a systemic fibro-inflammatory disease with multiple organ involvement.The mostly common involved organs include lymph node,submandibular glands,and pancreas.Glucocorticoids and immunosuppressive agents were effective for IgG4-RD.

Objective To explore the effect of enhanced recovery after surgery (ERAS) in reducing post-traumatic stress disorder (PTSD).Methods Totally 95 patients who received cardiopulmonary bypass surgery in the Department of Cardio-Thoracic Surgery, the Second People's Hospital of Liaocheng between January 2013 and July 2014 were selected as a control group, while another 95 patients who were hospitalized between August 2014 and December 2015 were selected as a treatment group. The patients in the control group received conventional nursing in the Department of Cardio-Thoracic Surgery, while the patients in the treatment group received ERAS on the basis of conventional nursing. The positive rate and level of PTSD symptoms of the patients in the two groups were then evaluated with PCL-C Scale.Results The positive rates of the patients in the treatment group and the control group were 3.16% and 10.53%, respectively (χ2=4.046,P<0.05). The PCL-C scores at different time were statistically different between the patients in the two groups(F=135.15,P<0.05),the PCL-C scores were statistically different between the patients in the two groups(F=33.84,P<0.05),and there were interaction between time and intervention (F=8.47, P<0.01). The positive rates of higher awareness, emphasis on experience and evasion/numbness symptoms of the patients in the treatment group were 21.89%, 12.42% and 10.83%, lower than those of the patients in the control group 28.42%, 15.16% and 14.73% (χ2=16.118, 4.489, 13.678;P<0.01).Conclusions The concept of ERAS used during the perioperative period of cardiac surgery can effectively reduce the incidence and level of PTSD symptoms.

Objective To study the compatible stability of the carbohydrate-electrolyte injection and commonly used vitamin-electrolyte injections. Methods By simulating clinical use of medicines,the carbohydrate-electrolyte injection and various vitamin-electrolyte injections were mixed respectively.The content of sodium acetate was measured by HPLC,and changes in appearance,pH value and insoluble particles of the injections were observed. Results At room temperature,the compatibility solutions showed no significant changes in appearance,pH value,the number of insoluble particles and the content of sodium acetate within 8 h. Conclusion The carbohydrate-electrolyte injection is compatible with commonly used vitamin-electrolyte injections,and the admixtures are stable within 8 h at room temperature.