Objective To explore the diagnosis of clinically suspicious von Willebrand disease(vWD)in a family and its pathogene-sis.Methods The pedigree information and the biological specimen were collected from the clinically suspected VWD patient and her family members(4 persons in total)in Peking University First Hospital.The levels of platelet count(PLT),activated partial thrombo-plastin time(APTT),vWF antigen(vWF:Ag),vWF activity(vWF:Ac)and FⅧ activity(FⅧ:C)were detected,and vWF risto-cetin cofactor(vWF:RCo)assay,ristocetin-induced platelet aggregation assay(RIPA)and vWF collagen binding(vWF:CB)assay were performed for phenotype diagnosis.The peripheral blood genomic DNAs were extracted from the proband and her family members to perform whole-exome sequencing for identifying the mutation of vWF gene,The mutation site was analyzed by using bioinformation tools to explore the pathogenesis of the proband.Results The APTT of proband(m 1)was slightly prolonged and her vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were significantly decreased.There was no obvious aggregation in RIPA assay(1.0 mg/mL and 1.25 mg/mL).In her father(Ⅱ3),APTT,FⅧ:C,vWF:Ag,vWF:Ac and vWF:CB were normal,but vWF:RCo was slightly decreased.In her mother(Ⅱ4),APTT,FⅧ:C,vWF:Ag,vWF:RCo and vWF:CB were all normal,but vWF:Ac significantly decreased.In her brother(Ⅲ2),APTT and FⅧ:C were normal,but vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were reduced to varying degrees.In all the family members(father,mother and brpther),no apparent aggregation in RIPA(1.0 mg/mL)was shown.Genetic analysis showed that the proband(Ⅲ1)carried a compound heterozygous mutation of vWF gene c.7288-9T＞G and c.1117C＞T,her father(Ⅱ3)carried vWF gene c.7288-9T＞G heterozygous mutation,and vWF gene c.1117C＞T heterozygous mutation was presented in both mother(Ⅱ4)and brother(Ⅲ2).Conclusion According to the results of laboratory tests,the proband was diagnosed as type 2A vWD.The hetero-zygous mutation in vWF gene c.1117C＞T and c.7288-9T＞G may be the molecular mechanism leading to type 2A vWD in the proband.

This paper summarized the clinical experience of Professor GE Linyi in treating ulcerative colitis （UC） by stages with the method of clearing. Professor GE believes that the core pathogenesis of UC is dampness and heat in the intestines， and by taking the method of clearing as the basis， he proposed four methods for treatment of UC including clearing and transforming， clearing and dispersing，clearing and moisterning， clearing and nourishing. The pathogenesis of UC in its active stage is dampness and heat in the intestines， congestion and stagnation of qi and blood， and accumulation of stasis toxins， for which the treatment method is to clear and transform， accompanied by clearing and dispersing method. In terms of the clearing and transforming method， Bai Tou Weng Decoction （白头翁汤） combined with Haungqin （Radix Scutellariae）， Machixian （Herba Portulacae） and Pugongying （Herba Taraxaci） is taken as the basic prescription to clear and transform dampness and heat， cool blood， resolve toxins and stop dysentery. For the clearing and dispersing method， medicinals to rectify qi such as Chaihu （Radix Bupleuri）， Cuxiangfu （Vingar Rhizoma Cyperi）， Muxiang （Radix Aucklandiae）， Zhiqiao （Fructus Aurantii）， and Binlang （Semen Arecae）， as well as those to regulate blood such as Danggui （Radix Angelicae Sinensis）， Cebaiye （Cacumen Platycladi） and Diyutan （Radix Sanguisorbae Carbonisatus） are recommended. The pathogenesis of the remission stage is healthy qi depletion and lingering pathogen of dampness and heat stasis toxin in the intestines， for which the method of clearing and nourishing， clearing and moistening can be used； the latter is mainly for people with yin fluids injury， and self-made Qingrun Yichang Decoction （清润益肠汤） is recommended， while the former is for those with spleen and stomach weakness， and self-made Qingyang Jianpi Decoction （清养健脾汤） can be used.

Objective:To explore the feasibility of a stable pig-to-monkey orthotopic liver transplantation (LT) model and provide a favorable experimental tool for preclinical studies of xenotransplantation.Methods:In this retrospective analysis, the authors reviewed the perioperative conditions and outcomes of 7 pig-to-monkey orthotopic liver transplants performed by a xenotransplantation research team of Second Xiangya Hospital.Gene-edited Banna miniature pigs were selected as donors and rhesus monkeys with similar anatomical characteristics, physiology, biochemistry and immune mechanism to humans were selected as recipients for pig-monkey xenogeneic orthotopic LT.Based upon classic transplantation procedures, whole liver xenogeneic orthotopic transplantation was performed.Surgical processes were modified for minimizing intraoperative hemorrhage and shortening anhepatic period.A bile duct drainage tube was implanted for observing bile secretion.ATG + anti-CD20 + snake venom factor and FK-506 were utilized for immunoinduction pre-operation while tacrolimus, mycophenolate mofetil (MMF) and methyl prednisolone for postoperative immunomaintenance therapy.Antibiotics and antiviral agents were also applied and thrombin complex for improving coagulation functions.Results:All procedures were successfully completed.After the stability and maturity of our model, in case No.7, donor's acquisition operative duration was 42 min without heat ischemic time, donor's trimming time 87 min, donor cold retention time 128 min, recipient's operative duration 123 min and anhepatic phase 27 min.Subhepatic inferior vena cava was occluded for 38 min.Blood loss was around 10 ml.And 4/7 models survived post-operation and the longest survival time was 27 h. Among 3 non-survival cases, the causes were anesthesia accident (n=1) and immaturity of early operation (n=2). Model No.7 had a biliary secretion volume of 86 ml post-operation.Conclusions:Qualified donor acquisition, high-quality vascular anastomosis, intraoperative reduction of blood loss, shortening of anhepatic period, strict fluid replenishing and careful monitoring are essential for boosting the success rate of pig-monkey liver xenotransplantation model.Optimization of donor gene combination and advanced immunosuppression protocol help to further achieve the long-term survival of pig-monkey liver xenotransplantation model.

Objective:To explore the effect of health management based on constitution identification in perimenopausal women.Methods:From November 2020 to April 2021, 244 perimenopausal women from Doumen Street, Yuecheng District, Shaoxing were selected as the research subject by convenience sampling method, and divided into the control group (121 cases) and the observation group (123 cases) according to their respective communities. The control group adopted routine health management, while the observation group implemented health management based on constitution identification on the basis of routine health management. Three months after intervention, the perimenopausal symptom scores between the two groups were compared. One year after intervention, the number of individuals with biased constitution and quality of life scores between the two groups were compared.Results:Three months after intervention, the scores of hot flashes, sweating, insomnia, dizziness, fatigue, muscle and joint pain, headache, palpitations, and total score of perimenopausal symptom of the observation group were lower than those of the control group, with statistically significant differences ( P<0.05). Three months after intervention, there were no significant differences in scores of paraesthesia, mood swings, depressive suspicion, skin ant sensation, sexual difficulties, urinary tract infection between the observation group and the control group ( P>0.05). One year after intervention, the number of individuals with biased constitution of the observation group was less than that of the control group, and the difference was statistically significant ( P<0.05). One year after intervention, the scores of physical health, mental health, social relationship, and total score of quality of life of the observation group were higher than those of the control group, and the differences were statistically significant ( P<0.05). One year after intervention, there was no statistically significant difference in the score of the surrounding environment between the observation group and the control group ( P>0.05) . Conclusions:Health management based on constitution identification can improve the perimenopausal symptoms of perimenopausal women, gradually change their biased constitution towards a calm constitution, and improve their quality of life.

Objective To investigate the efficacy and safety of programmed cell death protein-1 (PD-1) monoclonal antibody on the treatment of malignant tumor after solid organ transplantation (SOT). Methods The relevant literatures in 7 databases were searched. The data on 54 cases of recipients with malignant tumors treated with PD-1 monoclonal antibody after SOT were collected, and the clinical effects and rejection of SOT recipients treated with PD-1 monoclonal antibody were analyzed. Results Total 32 acceptable articles including 54 SOT recipients were incorporated, including 43 males and 11 females aged 14-79 years old. There are 29 renal transplant recipients, 19 liver transplant recipients and 6 heart transplant recipients. The types of PD-1 monoclonal antibody agent used by SOT recipients included pembrolizumab for 28 patients and nivolumab for 26 patients. The overall remission rate, disease progression rate and fatality rate of PD-1 monoclonal antibody for postoperative malignant tumors of SOT recipients were 32% (17/54), 44% (24/54) and 36% (19/54), respectively. After treatment with PD-1 monoclonal antibody for postoperative malignant tumors of SOT recipients, the incidence of rejection was 39% (21/54), indicating no significant correlation between rejection and type of PD-1 monoclonal antibody (P > 0.05). Conclusions PD-1 monoclonal antibody can effectively treat postoperative malignant tumors of SOT recipients, and may induce rejection during the treatment. But rejection is not the most common cause for death of recipients.

Objective To evaluate the effect of donor risk index (DRI) on the early prognosis of liver transplantation for acute-on-chronic liver failure (ACLF). Methods Clinical data of 159 ACLF recipients undergoing liver transplantation were retrospectively analyzed. According to the calculation formula of DRI, all recipients were divided into DRI < 1.65 group (n=96) and DRI≥1.65 group (n=63). Based on the Chronic Liver Failure Consortium acute-on-chronic liver failure score (CLIF-C ACLFs), all recipients were divided into CLIF-C ACLFs < 48 group (n=78) and CLIF-C ACLFs≥48 group (n=81). The early prognosis indexes including the length of intensive care unit (ICU) stay and the length of postoperative hospital stay of the recipients in each group were observed after liver transplantation. The 90 dsurvival rate of the recipients after liver transplantation was analyzed by Kaplan-Meier survival curve. The risk factors affecting the early prognosis of ACLF recipients after liver transplantation were analyzed by Cox's hazards regression model. Results The length of ICU stay and the length of postoperative hospital stay did not significantly differ between the DRI < 1.65 group and DRI≥1.65 group (both P > 0.05). The length of postoperative hospital stay did not significantly differ between the CLIF-C ACLFs < 48 group and CLIF-C ACLFs≥48 group (P > 0.05). The length of ICU stay in the CLIF-C ACLFs < 48 group was 4 (3-14) d, significantly shorter than 7 (1-33) d in the CLIF-C ACLFs≥48 group (P < 0.05). The CLIF-C ACLFs was a risk factor of the early prognosis of ACLF recipients after liver transplantation (P < 0.05). The postoperative 90 d survival rate did not significantly differ between the DRI < 1.65 group and DRI≥1.65 group (P > 0.05). The postoperative 90 d survival rate in the CLIF-C ACLFs < 48 group was 94%, significantly higher than 79% in the CLIF-C ACLFs≥48 group (P < 0.05). Conclusions The early prognosis of ACLF recipients after liver transplantation is correlated with the severity of the disease rather than the DRI. Liver transplantation should be performed early and promptly.

Objective To evaluate the outcome of 1iver transplantation for acute-on-chronic liver failure (ACLF) patients .Methods We included 453 consecutive patients with previously cirrhosis who underwent liver transplantation between January 2013 and December 2017 .Patients were categorized as no ACLF (n=294) and ACLF(n=159) according to EASL-CLIF consortium criteria .Furthermore ,we used ACLF grades to categorize the ACLF patients .Their clinical data were reviewed and their 90-days survival outcomes were compared .Results Compared with the no ACLF group ,the length of stay in the ICU was significantly prolonged for all patients with ACLF ,and the 90-days survival rate after transplantation was significantly reduced in ACLF group .The length of stay in the ICU was shorter in Grade 1 and Grade 2 group when compared to Grade 3 group .The 90-days survival rate of no ACLF ,Grade 1 ,Grade 2 and Grade 3 group were 93 .20％ ,92 .59％ ,93 .33％ and 73 .68％ ,respectively .There were no statistically significant differences in 90-days survival rate among the no ACLF ,Grade 1 and Grade 2 group .However , the 90-days survive rate of Grade 3 group was lower than that of other groups .Conclusions Liver transplantation has been shown to be safe and effective with good outcome in patients with ACLF and should be offered in early course of ACLF before onset of multi-organ failure .

Objective To construct the orthotopic mouse liver transplantation model and cover troubleshooting,in order to provide experimental techniques support for organ transplantation pathology and immunology studies.Methods Male C57BL/6 mice,10-12 weeks,were selected as the allograft donors.Male C3H mice with same age were selected as the allograft recipients.The orthotopic mouse liver transplantation model consisted of 3 stages,including harvesting the donor liver,back-table preparation of the liver graft and transplantation of the donor liver into the recipient.The average time for harvesting the donor livers was (40 ± 8.8) min,(23 ± 4.7) min for preparing the donor livers and (75 ± 9.6) min for transplanting the donor livers into the recipient.Results Seventy pairs of mice were used for the preliminary experiments.For the formal experiments,the allograft transplantation was established on 220 pairs with 90.4％ successful rate.Conclusion It is the skillful and high quality microsurgical technique that is the guarantee of establishing the orthotopic mouse liver transplantation model successfully.

Objective To observe the expression of aquaporin 4 (AQP-4) in contused brain tissue and its relationship with brain edema following brain trauma.Methods A retrospective case control analysis was made on 42 patients with severe brain trauma admitted from January 2015 to March 2016.There were 23 males and 19 females,aged from 23 to 62 years [(35.5 ± 5.6) years].Glasgow coma score (GCS) was 3-5 points in 7 patients,6-8 points in 23 and 9-10 points in 12.Brain tissue removed from the area 1 cm near the contusion during the cranial surgery were allocated to study group (n =42),while brain tissue removed far from the contusion after internal decompression were used as control (n =8).Ultrastructure of brain tissues was observed under electron microscope.Water content of brain tissue was measured by dry-wet weight method and expression of AQP-4 was measured by immunohistochemical method at postinjury hours ＜ 6,6-12,12-24,24-72,72-96 and ＞ 96.Results Morphology and structure of brain tissue in control group were normal.Whereas in study group,the intracellular and interstitial edema were obvious and morphological structure were damaged.Water content and AQP-4 expression in control group showed no obvious increase after operation(73.55 ±0.10,0.193 ±0.016).Water content in study group increased significantly compared to control group and reached the peak value (81.28 ± 0.56) at postinjury 24-72 hours (P ＜ 0.01).AQP-4 expression in study group increased at postinjury 6 hours (0.242 ±0.023) and reached a peak at postinjury 24-72 hours (0.338 ± 0.013),with significant difference compared to control group (P ＜ 0.05).Correlation analysis showed change of brain water content was positively correlated with expression level of AQP-4 (r =0.931,P ＜ 0.01).Conclusion Expression of AQP-4 in the injured area of brain trauma is significantly increased along with the increase of water content,suggests that the upregulation of AQP-4 plays an important role in traumatic brain edema.

ObjectiveTo investigate the expression of T helper (Th) 17 cells and the related interleukin 17 (IL-17) in acute renal allograft rejection in mice and its significance.Methods We established a mouse renal allograft model,in which mice were randomly divided into a renal isograft group and an acute renal allograft rejection group.Three and 7 d after the transplantation,the serum interferon (IFN)-γand IL-17 levels in the mice were determined by enzyme-linked immunosorbent assay,the percentage of Th1 and Th17 cells in the total kidney-infiltrating lymphocytes was investigated by flow cytometry,and the transplanted kidney species were given routine pathological examination after fixation with 10％ formalin.ResultsCompared with the isograft group,the allograft mice showed a significantly higher content of IL-17 (P ＜0.05 ) but not IFN-γ in the serum 3 d after transplantation,and showed significantly higher serum IL-17 and IFN-γcontents 7 d after transplantation (P ＜ 0.05 ).Also,compared with the isograft group,the allograft mice exhibited significantly higher percentage of Th1 and Th17 cells on both day 3 and day 7 ( P ＜ 0.05 ).In the allograft group,the contents of serum IFN-γand IL-17 and the percentage of Th1 and Th17 cells were significantly higher on day 7 than on day 3 (P ＜ 0.05 ).Routine pathological examination indicated that,as time passed,the allograft mice showed gradually stronger rejection responses.ConclusionTh17 cells might play an important role in the development of acute renal allograft rejection,and IL-17 can be used as an early indicator of acute rejection.