AIM: To explore interaction and biological behaviou r changes of two kinds of cells-blastocysts and hepatocarcinoma cells in the same microenvi ronment. METHODS:The models of mouse blastocysts co-cultured wit h human hepatoca rcinoma cell lines were established, then biological behaviours and mutual effe c ts of the two kinds of cells in co-culture system were observed. RESULTS: Co mpared with control group, hepatocarcinoma cells with differently invasive and met astatic potential significantly enhanced the rates of blastocyst hatchment , at t achment and outgrowth(P0 05). The blastocyst ha tched and attached to hepatocarcinoma cells with differently invasive and metast atic p otential. Then, differential trophoblasts invaded hepatocarcinoma cells. The clear-cut interfaces were gradually formed between both sides. Hepatocarcinoma cells o n interface showed changes of growth direction and cell shapes and did not inv ade blastocysts. CONCLUSIONS: Hepatocarcinoma cells promoted bla stocyst develo pment. Blastocysts implanted and invaded hepatocarcinoma cells with differentl y i nvasive and metastatic potential in vitro, which indicate that blastocyst i mplan tation in vitro does not relate with the kinds and differential level of int erac tional cells and the low selectivity maybe relate with high adaptability of earl y life.

Objective To compare the change features of anti-donor specific antibody (DSA) in different species of sentitized mice after skin transplantation. Methods All mice were divided into the Balb/c → C57BL/6 (6 pairs) and Balb/c → C3H skin transplantation groups (6 pairs). At d0, d2, d4, d7, d13, d17, d28, d35, d42, d49 and d56 after skin transplantation, the serum sample was prepared for detection of DSA-IgG and DSA-IgM levels. Results Moderate increase was noted in the DSA-IgG level in the sensitized mice within 1 week after skin transplantation. The IgG level was significantly increased within 1-4 week and peaked and stabilized within 4-8 week. No significant variation was observed in the DSA-IgM level at 8 weeks after skin transplantation. In the Balb/c → C57BL/6 skin transplantation group, the DSAIgG level was significantly lower than that in the Balb/c → C3H group. Statistical significance was identified in the IgG levels between two groups at d2, d17, d28, d35, d42, d49 and d56 after skin transplantation (all P<0.05). No statistical significance was noted in the DSA-IgM levels between two groups at each time point (all P>0.05). Conclusions Advancing the time of renal transplantation after skin transplantation moderately in the Balb/c → C3H group, or changing to the lower immunoreactive combination of Balb/c → C57BL/6 are aimed to establish AMR mouse models with mild rejection reaction.

To investigate the diversity and composition of gut microbiota in patients with lung adenocarcinoma and lung squamous cell carcinoma. A single-center and case-control study was conducted to consecutively enroll a total of 27 lung cancer patients, including 15 males and 12 females, who were seen at the Affiliated Hangzhou First People′s Hospital, Zhejiang University School of Medicine between September 2018 to October 2020. A total of 20 cases of healthy healthy physical examiners, including 9 males and 11 females were recruited as healthy control group (HC) during the same period. Clinical data and stool samples were collected from each participants, and lung cancer patients were divided into lung adenocarcinoma group (AC, 19 patients, 8 males and 11 females) and lung squamous cell carcinoma group (SCC, 8 patients, 7 males and 1 females) according to the pathology type. Genomic DNA were extracted to amplify 16S rDNA V3-V4 region, then the Illumina MiSeq high-throughput sequencing platform and QIIME software were used for sequencing and analyzing the structure of the gut microbiota, respectively. Analysis of variance, χ 2 test, K-W test were used to analyze the differences in age, gender,α diversity, and relative abundance of microbiota among the three groups. AC, SCC, and HC were aged (58.74±9.27), (63.38±6.12), and (55.65±7.79) years old, respectively. There were no difference in gender and age among the three groups (gender and age are respectively:χ 2=5.155, P=0.076; F=2.598, P=0.086). And no significant difference in alpha diversity were found among the three groups (Chao and Shannon index were respectively: F=0.616, P=0.545; F=2.484, P=0.095), while β-diversity analysis indicated significant differences in the structure of intestinal flora among AC, SCC and HC ( P=0.001). LEfSe analysis showed that AC and SCC both have dominant bacterials. Megasphaera ( H=7.855, P=0.020) and Erysipelatoclostridium ( H=7.426, P=0.024) were enriched in patients with AC, while Enterococcus ( H=8.400, P=0.015), Veillonella ( H=9.957, P=0.007), and Eubacterium_eligens_group ( H=10.514, P=0.005) were enriched in patients with SCC. Lung cancer patients have gut microbiota imbalance, while lung adenocarcinoma and lung squamous cell carcinoma patients have no significant difference in gut microbiota diversity, but lung adenocarcinoma and lung squamous cell carcinoma have their own unique microbiota. This imbalance of the intestinal microenvironment is of great significance for studying the occurrence and development of different pathological types of lung cancer.

To investigate the diversity and composition of gut microbiota in patients with lung adenocarcinoma and lung squamous cell carcinoma. A single-center and case-control study was conducted to consecutively enroll a total of 27 lung cancer patients, including 15 males and 12 females, who were seen at the Affiliated Hangzhou First People′s Hospital, Zhejiang University School of Medicine between September 2018 to October 2020. A total of 20 cases of healthy healthy physical examiners, including 9 males and 11 females were recruited as healthy control group (HC) during the same period. Clinical data and stool samples were collected from each participants, and lung cancer patients were divided into lung adenocarcinoma group (AC, 19 patients, 8 males and 11 females) and lung squamous cell carcinoma group (SCC, 8 patients, 7 males and 1 females) according to the pathology type. Genomic DNA were extracted to amplify 16S rDNA V3-V4 region, then the Illumina MiSeq high-throughput sequencing platform and QIIME software were used for sequencing and analyzing the structure of the gut microbiota, respectively. Analysis of variance, χ 2 test, K-W test were used to analyze the differences in age, gender,α diversity, and relative abundance of microbiota among the three groups. AC, SCC, and HC were aged (58.74±9.27), (63.38±6.12), and (55.65±7.79) years old, respectively. There were no difference in gender and age among the three groups (gender and age are respectively:χ 2=5.155, P=0.076; F=2.598, P=0.086). And no significant difference in alpha diversity were found among the three groups (Chao and Shannon index were respectively: F=0.616, P=0.545; F=2.484, P=0.095), while β-diversity analysis indicated significant differences in the structure of intestinal flora among AC, SCC and HC ( P=0.001). LEfSe analysis showed that AC and SCC both have dominant bacterials. Megasphaera ( H=7.855, P=0.020) and Erysipelatoclostridium ( H=7.426, P=0.024) were enriched in patients with AC, while Enterococcus ( H=8.400, P=0.015), Veillonella ( H=9.957, P=0.007), and Eubacterium_eligens_group ( H=10.514, P=0.005) were enriched in patients with SCC. Lung cancer patients have gut microbiota imbalance, while lung adenocarcinoma and lung squamous cell carcinoma patients have no significant difference in gut microbiota diversity, but lung adenocarcinoma and lung squamous cell carcinoma have their own unique microbiota. This imbalance of the intestinal microenvironment is of great significance for studying the occurrence and development of different pathological types of lung cancer.

This report described one human leukocyte antigen pre-sensitized recipient undergoing preoperative plasmapheresis (PP), intravenous immunoglobulins (IVIG) desensitization and immune induction therapy before kidney transplantation with a donor kidney. Early postoperative clinical diagnosis was acute antibody-mediated rejection (AMR). A marked elevation of donor-specific antibodies (DSA) was accompanied by a decline of renal function. PP/IVIG dosing failed to lower the level of DSA. After low-dose splenic irradiation, DSA level dropped steadily and transplanted kidney function normalized. Thus adjuvant low-dose splenic irradiation may eliminate DSA immediately without a rebound.

Objective:To summarize the clinical characteristics and treatment of cytomegalovirus(CMV)infection in pediatric kidney transplant patients.Methods:From May 2014 to July 2021, a total of 9 cases(8.65%)of 104 pediatric kidney transplant recipients were diagnosed with CMV infection in our centre.Retrospective data was collected for these 9 paediatric recipients.The clinical characteristics of the disease, treatment data and outcomes were summarized.Results:The median age of the 9 children was 10 years(0.25-15 years), 6 of whom were treated with polyclonal antibody for immunity induction.CMV IgG was negative in 4 children before renal transplantation.Only one patient received anti-CMV prophylaxis.The median time from transplant to the diagnosis of CMV infection was 22(7-15)days.Among the 9 children, 7 had fever, pneumonia and diarrhea, 2 had no typical symptoms, three patients were complicated with viral, bacterial or fungal infections.Acute rejection occurred in 3 patients at the same time as CMV infection or after CMV DNA turned negative.Nine patients were cured and discharged after ganciclovir or valganciclovir treatment.Median time of CMV DNA negative transformation was 32(17-90)days.Conclusions:Pediatric transplant recipients are at particularly elevated risk of CMV disease.Antiviral prophylaxis should be initiated early after transplantation.

Objective:To summarize the clinical characteristics and therapeutic drug selection of post-transplantation diabetes mellitus(PTDM)after kidney transplantation in children.Methods:From May 2014 to March 2021, a total of 5 cases(5.38%)of 93 paediatric kidney transplant recipients with a median follow-up period of 34 months were diagnosed with PTDM in our centre.Retrospective data analysis was performed for these 5 paediatric recipients.The characteristics of the disease, treatment data and outcomes were summarized.Among the five paediatric recipients, one was male and four patients were female, ranging the age from 12 to 17 years.All recipients received a tacrolimus-based immunosuppressive regimen with prednisone discontinued no later than 3 months after kidney transplant.Results:The onset of PTDM ranged from 1 month to 46 months(median: 17 months)after transplantation.The blood glucose of two children returned to normal gradually after tacrolimus conversion to cyclosporine, with one of them was given insulin temporarily.Three children received oral hypoglycaemic agents, including one received acarbose, one received metformin, and one received metformin combined with acarbose.After a median follow-up of 6 months, the levels of blood glucose in five children were stable, and there was no significant change in serum creatinine and urine protein.Conclusions:The treatment of PTDM in children should be individualized with considering of age, gender and immunosuppressive regimen. Switch from tacrolimus to cyclosporine is effective. Metformin or other hypoglycemic agentsis helpful when tacrolimus is maintained.

Objective To investigate the safety,efficacy,and clinical value of a fracture reduc-tion and fixation system.Methods The design and application of components[Kirschner wire(K-wire)limiter and K-wire guiding locking device]of the fracture reduction and fixation system were described.Six surgeons implanted K-wires into synthetic bones using the K-wire limiter or by percep-tion respectively,and the length of K-wire protrusion was assessed.K-wire guiding locking device combined with locking plates and two 1.5 mm K-wires,two 1.5 mm K-wires,as well as two 2.0 mm K-wires were used to fix a fracture model constructed by synthetic bones,and the torque at failure was measured.Results The K-wire limiter reduced the length of K-wire protrusion(P＜0.001).The failure torque of fractures fixed by K-wire guiding locking device with locking plates and two 1.5 mm K-wires was larger than those fixed with two 1.5 mm K-wires and two 2.0 mm K-wires(P＜0.05).Conclusion The fracture reduction and fixation system offers a new technique for reducing and recon-structing fractures,assessing and optimizing the position of locking plate screws,as well as protecting the blood supply of fractures.

Objective To investigate the safety,efficacy,and clinical value of a fracture reduc-tion and fixation system.Methods The design and application of components[Kirschner wire(K-wire)limiter and K-wire guiding locking device]of the fracture reduction and fixation system were described.Six surgeons implanted K-wires into synthetic bones using the K-wire limiter or by percep-tion respectively,and the length of K-wire protrusion was assessed.K-wire guiding locking device combined with locking plates and two 1.5 mm K-wires,two 1.5 mm K-wires,as well as two 2.0 mm K-wires were used to fix a fracture model constructed by synthetic bones,and the torque at failure was measured.Results The K-wire limiter reduced the length of K-wire protrusion(P＜0.001).The failure torque of fractures fixed by K-wire guiding locking device with locking plates and two 1.5 mm K-wires was larger than those fixed with two 1.5 mm K-wires and two 2.0 mm K-wires(P＜0.05).Conclusion The fracture reduction and fixation system offers a new technique for reducing and recon-structing fractures,assessing and optimizing the position of locking plate screws,as well as protecting the blood supply of fractures.

Objective:To explore the efficacy and safety of a novel Tripterygium preparation (Kunxian Capsules)in kidney transplant recipients developing refractory proteinuria after transplantation.Methods:A total of 59 kidney transplant recipients received regular follow-ups from August 2018 to July 2021.Severity of proteinuria, kidney graft function and adverse effects were retrospectively recorded before and at Month 1/2/3/6 months after Kunxian treatment.They were divided into two groups of effective and void to explore the potential effect-related factors.Results:Six-month treatment was completed in 57 patients except for 2 cases of discontinued treatment due to severe adverse effects.A significant reduction in amount of proteinuria was observed at Month 1 [1.09(0.42, 2.59 g/24 h)vs 1.82(1, 2.7 g/24 h, P<0.01)]and the trend continued during subsequent follow-ups.Twenty-nine patients(50.9%)responded remarkably.Eighteen patients(31.6%)showed no response.The overall effective rate was 68.4%.Inter-group comparison revealed strong correlations between treatment efficacy and baseline serum creatinine levels, early initiation of treatment and symptom duration from onset to treatment.Kidney graft IgA nephropathy demonstrated the highest effective rate(11/13, 84.6%). Furthermore, elevated blood concentration of tacrolimus hinted at a potential drug interaction. Conclusions:Kunxian Capsules is efficacious and safe for renal transplant recipients developing moderate-severe proteinuria and not responding to traditional medications.Early initiation of treatment before graft function decline is recommended.Reducing tacrolimus dose cautiously in advance and monitoring adverse events are also essential.