BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.

Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K⁺ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β₂-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

Objective:To establish a sequencing method for the genome of severe fever with thrombocytopenia syndrome virus (SFTSV) based on next-generation sequencing (NGS).Methods:SFTSV RNA was extracted from serum samples of patients with severe fever with thrombocytopenia syndrome. SFTSV-specific primers were designed using Primer 5.0 software. A multiplex PCR method was constructed and used to amplify the nucleotide sequence of SFTSV. Whole-genome sequencing was performed on the NGS platform.Results:The whole genes of SFTSV isolates in 28 serum samples were amplified by the multiplex PCR with a coverage over 94%. Sequencing and phylogenetic analysis of those strains revealed that the predominant strains ( n=20) belonged to genotype A, followed by genotypes B ( n=4) and E ( n=3). Conclusions:A high-throughput sequencing method for SFTSV based on multiplex PCR was established in this study. This method was characterized by high specificity and good quality and could improve the sequencing efficiency.

Objective : To investigate the predictive value of serum homocysteine(HCY) level on the risk of miscarriage in polycystic ovary syndrome(PCOS) patients undergoing frozen-thawed embryo transfer(FET). Methods: 536 PCOS patients and 550 non-PCOS patients with fallopian tube factors(control group) retrospectively were analyzed.The basic characteristics and pregnancy outcomes were compared between the two groups.Also, the basic characteristics between the miscarriage group and the live birth group in PCOS patients were compared.The predictive value of Hcy on the miscarriage of PCOS patients after FET treatment was analyzed. Results: Compared with the control group, body mass index(BMI),testosterone(T),Hcy, fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),total cholesterol(TC),number of retrieved oocytes and miscarriage rate increased in PCOS group, while high density lipoprotein-cholesterol(HDL-C),high-quality embryo rate and live birth rate decreased, and the differences were statistically significant(P<0.05).Among PCOS patients, Hcy, FINS and HOMA-IR increased in the miscarriage group compared with live birth group, and the differences were statistically significant(P<0.05).Hcy, FINS and HOMA-IR were independent risk factors for miscarriage in PCOS patients, and the predictive value of Hcy for miscarriage in PCOS patients was greater than FINS and HOMA-IR,and the area under ROC curve(AUC) was 0.829(95%CI=0.755-0.902,P<0.001),the cut-off value was 15.975 μmol/L with a 79.0% sensitivity and a 87.4% specificity, which indicated high predictive value for miscarriage in patients with PCOS. Conclusion Serum Hcy level significantly increased in PCOS patients, which has an important predictive value for miscarriage after FET treatment.

Objective:To explore the effect of kangaroo mother care (KMC) on lactation, uterine involution of parturients and neonatal pain.Methods:A total of 200 parturients who gave birth at full term in our hospital from January 2019 to June 2019 and their newborns were selected as the research objects, they were randomly divided into control group and observation group of 100 pairs each. The control group received routine obstetric postpartum care and the observation group received KMC. The KMC cognition, postpartum lactation and uterine involution, neonatal pain during neonatal heel blood collection were compared between the two groups.Results:The cognition of KMC in the observation group was significantly better than that in the control group, and the difference was statistically significant ( χ 2 value was 24.700, P<0.01). The first lactation time of parturients in the observation group was (41.25±3.20) hours after birth, which was earlier than (54.17±2.20) hours in the control group, there was significant difference between the two groups ( t value was 2.378, P value was 0.019). The breast pain Ⅰ degree (20 cases), Ⅱ degree (56 cases), Ⅲ degree (24 cases) in observation group were significantly lighter than those in control group (62, 27, 11 cases respectively) 72h after delivery, the differences were statistically significant ( t value was 12.166, P value was 0.011). The parturients of sufficient lactation in the observation group (73 cases) were more than those in the control group (34 case),the differences was statistically significant ( χ 2 value was 30.570, P value was 0.000). The uterine fundus of the observation group decreased by (3.06±1.26) cm and (1.67 ±0.43) cm at 24h and 48h postnatally, which were better than those of the control group (1.97±0.92) cm and (1.23±0.18) cm,the differences were statistically significant ( t value was 3.162, P value was 0.002; t value was 2.689, P value was 0.009). In the process of heel blood collection after 72h of delivery in both groups, the pain scores of the observation group during and after blood collection were 4.92±0.33 and 2.37±1.27 respectively, which were lower than those of the control group (5.57±1.37 and 5.01±1.09), and the differences were statistically significant ( t value was 2.035, P value was 0.046; t value was 2.579, P value was 0.011). The heart rates of the observation group during and after blood collection were (121.36±22.13) and (142.55±23.91) beats/min, respectively, which were lower than (152.64±18.21) and (156.79±17.37) beats/min of the control group, the difference were statistically significant ( t value was 2.375, P value was 0.018; t value was 2.126, P value was 0.037). The blood oxygen saturation of the observation group during and after blood collection were 0.967 2±0.013 7 and 0.985 5 ±0.022 4 respectively, which were significantly higher than 0.891 7±0.116 5 and 0.914 5±0.137 8 of the control group ( t value was 2.036, P value was 0.046; t value was 2.017, P value was 0.047). Conclusions:The implementation of KMC can promote lactation, accelerate uterine involution, and relieve the pain of neonats during neonatal heel blood collection; Strengthening the health education of KMC can improve the cognition of parturients and their families about KMC, which has positive significance in promoting maternal and infant health and is worthy of clinical application.

Objective To investigate the diagnostic effect of diffusion tensor imaging (DTI) on spinal neurotype brucellosis spondylitis (BS).The characteristics of apparent diffusion coefficient (ADC) value and fractional anisotropy (FA) value of spinal neurotype BS in different disease stages were quantitatively analyzed to evaluate the different forms of spinal neurofibrillary tract injury.Methods A prospective design was used to collect data of BS patients with spinal neurological symptoms from June 2015 to July 2017 in the First Affiliated Hospital of Hebei North University as the brucellosis group (n =39),including 23 males and 16 females,aged (20.8 ± 15.3) years old.Healthy subjects were selected as the control group (n =30),including 20 males and 10 females,aged (25.2 ± 4.0) years old.The brucellosis group was divided into acute stage (＜ 3 months),subacute stage (3-6 months) and chronic stage (＞ 6 months),with 12,10 and 17 cases,respectively.Routine spinal scans and DTI scans were performed using a 3.0T superconducting magnetic resonance imaging (MRI) scanner,DTI used Fiber Trak package to measure ADC value and FA value and perform quantitative analysis [receiver operating characteristic (ROC) curve],and reconstructed the changed form of the spinal neurotype BS nerve fiber bundle.Results A total of 39 patients' data were collected in the brucellosis group.Among them,5 patients showed segmental thickening of spinal nerves on conventional MRI,high signal on T2 weighted imaging (T2WI) and short time inversion recovery (STIR),and color code changes on DTI scan FA imaging.Routine MRI of 34 patients showed spinal cord compression,spinal cord morphological changes or cauda equina nerve aggregation and displacement,while DTI scan FA imaging showed spinal cord or cauda equina nerve morphological changes,but no color code changes.The ADC values of patients in the acute and subacute stages were higher than that of the control group [(1.41 ± 0.05),(1.31 ± 0.05),(1.23 ± 0.05) × 10-3 mm2/s,P ＜ 0.05],and the FA values were lower than that of the control group (0.40 ± 0.04,0.68 ± 0.08,0.76 ± 0.05,P ＜ 0.05).There were no statistically significant differences in ADC and FA values between patients with chronic spinal neurotype BS [(1.25 ± 0.04) × 10-3 mm2/s,0.72 ± 0.04] and the control group (P ＞ 0.05).ROC curve analysis showed that the area under curve (AUC),specificity,sensitivity and accuracy of ADC were 0.912,0.942,0.930 and 0.924,respectively.The AUC,specificity,sensitivity and accuracy of FA were 0.901,0.937,0.928 and 0.943,respectively.The change forms of spinal neurotype BS were:① color code change;② loss/fracture;③ displacement and pressure;④ rarity.Conclusion DTI plays a diagnostic role in spinal neurotype BS,and can quantitatively analyze the characteristics of changes in ADC value and FA value in different periods,and can clearly display the forms of changes in spinal neurofiber,providing a reliable basis for the diagnosis of spinal neurotype BS.

Objective: To investigate the clinical effects of free superficial femoral artery femoral triangle perforator flap in the repair of skin and soft tissue defects in extremities. Methods: From January 2016 to November 2017, 14 patients (9 males and 5 females, aged 19 to 54 years) with skin and soft tissue defects in extremities accompanied with tendon and bone exposure were admitted to our unit. The size of skin and soft tissue defects after debridement ranged from 7 cm×3 cm to 10 cm×7 cm. The defects were repaired with free superficial femoral artery femoral triangle perforator flaps, with size ranging from 13.0 cm×2.0 cm to 20.0 cm×4.5 cm. The medial femoral cutaneous nerve was applied to the flap. The perforator flap was grafted onto the medial femoral cutaneous nerve in 6 patients. The donor sites were sutured directly. The survival of flaps and the follow-up of patients were observed. Results: All flaps of 14 patients survived successfully. The recipient sites and donor sites were healed completely in 13 patients, and 1 patient with partial skin necrosis at the edge of flap was healed after treatment. All patients were followed up for 6 months to 1 year after the operation. The flaps were in good shape, with nearly normal color and soft texture and no cicatrix contracture deformity. The flaps recovered protective sense in 6 patients who had medial femoral cutaneous nerve grafting, and the sensory recovery of the flap was slightly worse in the remaining 8 patients. There was no significant complications on the appearance and walking of the donor thigh in 14 patients, only a linear scar was left on the inner thigh, and no numbness was felt in the donor sites of patients. Conclusions The free superficial femoral artery femoral triangle perforator flap is an ideal therapy for repairing skin and soft tissue defects in extremities.

Objective To explore the differential diagnosis of breast ductal carcinoma in situ (DCIS)and breast ductal carcinoma in situ with microinvasion (DCIS-Mi)by ADCMin ,ADCDR and DCE-MRI,and to analyze the correlation between DCIS-Mi and biological factors. Methods Preoperative breast MRI examinations were performed in 41 patients with DCIS-Mi and 3 7 patients with DCIS.DCIS-Mi and DCIS patients were compared in terms of ADCMin ,ADCMax ,ADCDR ,early enhancement rate (EER)and the morphological characteristics of DCE-MRI.The optimal diagnostic variables were determined by binary Logistic regression,the threshold value of the optimal diagnostic variables was ensured by ROC,and the correlation between DCIS-Mi and biological factors was analyzed by Spearman.Results ADCMin of DCIS-Mi patients was lower than that of DCIS (t＝6.294,P＝0.033),and ADCDR was higher than that of DCIS (t＝9.246,P＝0.020).70.7 3% DCIS-Mi showed non-tumor-like enhancement,inclined to segmental distribution,and internal heterogeneous or cluster ring enhancement;29.27% manifested tumor-like enhancement,internal heterogeneous or ring enhancement,and unclear margin.64.86% DCIS showed non-tumor-like enhancement,inclined to linear distribution,internal homogeneous/heterogeneous enhancement;35.14% expressed tumor-like enhancement,internal homogeneous enhancement,and clear margin.The accuracy,sensitivity and specificity of ADCMin , ADCDR ,tumor or non-tumor internal enhancement features in the diagnosis of DCIS-Mi were higher (84.0%,9 5.3%,9 2.4%;89.3%, 9 5.3%,9 2.4%;85.1%,9 2.5%,9 3.8%;87.4%,9 6.8%,84.7%, respectively).ADCMin and ADCDR threshold value were 1.1 1× 10－3 mm2/s and 0.35×10－3 mm2/s,respectively.ADCMin of patients with DCIS-Mi was positive correlation with ER(－)and PR(－), and negative correlation with HER-2(+)(P＜0.05).ADCDR ,non-tumor distribution,and non-tumor internal enhancement characteristics,the tumor edge and internal enhancement characteristics were negative correlation with ER(－)and PR(－),and positive correlation with HER-2 (+)(P＜0.05).Conclusion ADCMin ,ADCDR and DCE-MRI can be used for the differential diagnosis of DCIS-Mi and DCIS, and provided evidence for clinical treatment plan.

Objective: To observe the effects of music electro-acupuncture and pulsed electro-acupuncture on locomotor avtivity and hippocampal astrocytes in fast-aging (SAMP8) mice, and to compare the anti-dementia mechanism.Methods: Thirty 8-month-old male SAMP8 mice were randomly divided into model group (group M), music electroacupuncture group (group MA), pulsed electro-acupuncture group (group EA) (n=10), with homologous normal aging SAMR1 mice as control group (group C) (n=10) . Acupuncture stimulation was applied to"Baihui" (GV20) 、"Yintang" (GV29) and"Renzhong" (DU26) for 20 min per day for 15 days. The Morris water maze was used to assess learningmemorize ability. Immunohistochemical DAB staining was used to observe GFAP exptession in hippocampus. Detect GFAP protein levels by Western blot in hippocampus. Results: The Morris water maze test showed: Compared with group C, the escape latency were increased (P ＜ 0.01), swimming distance were increased in space exploration experiment in group M (P ＜ 0.01) . Compared with group M, the escape latency were reduced in group MA and EA (P ＜ 0.01, P ＜ 0.05) and the swimming distance were shortened (all P ＜ 0.05) . The immunohistochemical DAB staining showed: Compared with group C, the average optical density of GFAP in the hippocampal CA1 area of group M increased significantly (P ＜0.01); the optical density of GFAP in CA1 area of hippocampus in both groups MA and EA was significantly decreased (all P ＜ 0.01), while the optical density of GFAP in MA group was lower than that of EA group (P ＜ 0.05) . Western blot test showed: Compared with group C, the expression of GFAP protein obviously increased (P ＜ 0.01) . Compared with group M, the expression of GFAP protein obviously decreased and had a downregulation tendency in group MA and EA (all P ＜ 0.05), while the expression of GFAP protein decreased in MA group compare with EA group (P ＜ 0.05) .Conclusion: Both music electro-acupuncture and pulsed electro-acupuncture can improve the learning and memory abilities of SAMP8 mice and decrease the expression of GFAP, which may be related to its mechanism of reducing neuroinflammatory reaction, improving apoptosis and eventually protecting neurons, and music electro-acupuncture has a better tendency than pulsed electro-acupuncture.

Objective To explore the role of cell apoptosis pathway in alcoholic pancreatitis.Methods C57BL/J mice were divided into control group (NC) and Alcohol group (AC),Acute pancreatitis group (AP) and Alcoholic acute pancreatitis group(AAP).Alcohol treatment was 10％ w/v ethanol feeding for 2 d,15％ w/v ethanol for 5 d,and then 20％ w/v ethanol until 13 weeks.AP model was established by the intraperitoneal injection of 50μg caerulein/kg body weight once an hour for a total of 7 times.Blood samples were collected for detecting serum amylase and lipase activity.Part pancreatic tissue was collected and the wet and dry weight were both measured to calculate the water content.The routine pathological exanination of the pancreatic tissues were conducted.The expression of apoptosis associated protein caspase3 and caspase8 was determined by Western blot.And cell apoptosis was determined using TUNNEL method.Results The level of serum amylase in NC group,AC group,AP group and AAP group were(3 630 ± 259),(3 196 ± 187),(35 955 ± 4607) and (53 607 ± 3 848) U/L;the level of serum lipase were (502 ± 41),(745 ± 42),(7 346 ± 665) and(12 764 ± 2 544) U/L;the water content were (70.2 ± 3.1) ％,(69.6 ± 2.0) ％,(78.2 ± 1.5) ％ and(85.0 ± 3.0) ％ and (12.75 ± 0.25);the expression of caspase3 were (1.017 ± 0.0784),(1.287 ± 0.097),(178 ± 0.07785) and (0.2443 ± 0.0243);the expression of caspase8 were (0.8289 ± 0.0096),(0.5985 ±0.0735),(1.27 ±0.08) and (0.145 ±0.015);the number of apoptotic cells were 1,6,214,97/10 high power field.The pathological score of pancreas injure in NC group,AC group,AP group and AAP group were 0,0,(7 ± 0.4) and (12.8 ± 0.3),respectively.Serum anylase,lipase,water content and pathological scores in AP group were obviously higher than those in NC group (P ＜ 0.05),which in AAP group were also obviously higher than those in AP group,and all the differences were statistically significant (all P ＜0.05).Compared with NC group,the expressions of apoptosis associated protein caspase3 and caspase8 and the number of apoptotic cells were obviously increased in AP group,which were obviously higher than those in AAP group,but the expression of caspase3 and caspase8 in AAP group were decreased compared with NC group,and all the differences were statistically significant (all P ＜ 0.05).Conclusions Chronic alcohol exposure may aggravate the severity of pancreatitis,and the inhibition of apoptosis pathway and the enhancement of acinar cell necrosis may be involved in this process.