Objective To investigate the effects and mechanism of lycopene on the proliferation and apoptosis of human hepatoma cell HepG2.Methods The human HepG2 cells in logarithmic growth phase were treated with lycopene (5,10,20 μg/ml) and cisplatin (40 pg/ml).The cellular growth inhibition rate was calculated by methyl thiazolyl tetrazolium (MTT) after 48 h,and cell cycle and apoptosis rate were analyzed by flow cytometry.The expressions of Bax,Bcl-2 and cleaved Caspase-3 proteins were detected by Western blotting.Results After 48 h intervention,the cellular growth inhibition rate of human HepG2 cell in blank control group was 0,and in lycopene (5,10,20 μg/ml) groups and cisplatin group were (21.3 ±4.2)％,(40.5 ±7.6) ％,(63.8 ± 9.1) ％,(37.8 ± 5.9) ％,with significant difference (F =37.905,P =0.000);and compared with blank control group respectively,the differences were statistically significant (t =208.124,P =0.000;t=394.637,P=0.000;t=628.592,P=0.000;t=257.168,P=0.000).The G0-G1 phase rates in lycopene (10,20 pg/ml) groups were (54.0 ± 2.9) ％ and (67.3 ± 3.6) ％,compared with blank control group (37.9± 1.5)％,the differences were statistically significant (t =4.508,P =0.024;t =10.673,P =0.006).The G2-M phase rates in lycopene (10,20 μg/ml) groups were (8.5 ± 0.6)％ and (4.7 ± 0.5)％,compared with blank control group (18.4 ± 0.8) ％,the differences were statistically significant (t =9.975,P =0.008;t =13.864,P =0.003).The apoptosis index (AI) in lycopene (5,10,20 μg/ml) groups and cisplatin group were (19.5 ± 4.8) ％,(43.0 ± 9.2) ％,(67.6 ± 10.1) ％ and (36.9 ± 6.8) ％,compared with blank control group [(3.6 ± 1.7)％],the differences were statistically significant (t =18.617,P =0.001;t =34.295,P =0.000;t =51.437,P =0.000;t =29.804,P =0.000).The expressions of Bcl-2,Bax and the ratio of Bax/Bcl-2 in lycopene (10,20 μg/ml) groups and cisplatin group were 0.42 ± 0.09,0.43 ±0.14,1.02 ±0.39;0.27 ±0.08,0.76 ±0.19,2.81 ±0.85;0.34 ±0.11,0.31 ±0.09,0.91 ±0.40,respectively.Compared with blank control group (0.59 ±0.17,0.18 ±0.06,0.31 ±0.12),the expressions of Bcl-2 were significantly down-regulated,and the differences were statistically significant (t =4.327,P =0.023;t =11.064,P =0.006;t =5.182,P =0.018),the expressions of Bax were significantly upregulated,and the differences were statistically significant (t =9.837,P =0.008;t =17.349,P =0.001;t =10.165,P =0.007),the ratios of Bax/Bcl-2 were significantly increased,and the differences were statistically significant (t=11.521,P=0.006;t=18.194,P=0.001;t=9.537,P=0.008).The expressions of cleaved Caspase-3 protein in lycopene (5,10,20 μg/ml) groups and cisplatin group were 0.25 ± 0.07,0.34 ±0.11,0.46 ± 0.18 and 0.17 ± 0.05,compared with blank control group (0.08 ± 0.03),the differenees were statistically significant (t =8.307,P =0.009;t =13.067,P =0.006;t =16.218,P =0.003;t =5.202,P =0.019).Conclusion Lycopene has inhibitive effect on the growth of human HepG2 cells perhaps through inhibiting proliferation and inducing apoptosis,which may be related to its effects of altering the cell cycle and the expressions of apoptosis-related genes and proteins.

BACKGROUND:With increased age,bone marrow mesenchymal stem cells(BMSCs)are influenced with regard to quantity and quality,which will induce great damages to the donors.Many studies have focused on seeking substitute MSC source.In contrast it remains controversial whether umbilical cord blood contains MSCs.OBJECTIVE:To isolate MSCs from human umbilical cord blood,and to detect their biological properties.METHODS:Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells.DMEM medium containing fetal bovine serum,penicillin,streptomycin and L-glutamine was added.Following several adherences and purification,the floating cells were discarded.Thus,many adherent cells with a confluence were collected.When cells were 60% 80% confluent,cells were digested by trypsin for subculture.Cells at passages,1,5 and 9 were obtained and their morphological changes were observed.Cell surface antigens were measured using flow cytometry.Growth curves were drawn,and cell viability was determined utilizing MTT.RESULTS AND CONCLUSION:Isolated umbilical cord blood MSCs presented an even size,showing spindle or star-shape fibroblasts-like cells.Umbilical cord blood MSCs at 1,5,9 passages were greatly positive for CD29,CD105 and CD166,but weakly positive for CD34 and CD45.Following 5 days of incubation,cells entered logarithmic growth phase.The number was decreased at day 9.Population doubling time was(53.5±8.32)hours.Cells grew well.Cells at 1-7 passages showed similar viability(P ＞ 0.05).Till passage 9,cell proliferation viability was decreased,but no significant difference was determined(P ＞0.05).Results verified that MSCs can be successfully isolated from umbilical cord blood in vitro.Cells at passages 1-9 presented a good reproductive activity.

Objective Cystic nephroma ( CN ) is an unusual benign neoplasm with high misdiagnosis rate , and at present there is no general method on its treatment .This study aimed to analyze the diagnosis and treatment of CN based on clinical data of CN patients. Methods We retrospectively analyzed the clinical data on 25 patients (including 2 cases of male pediatric patients , aged 14 and 16 years old, and the remaining 23 cases were adults, 11 males and 12 females, aged 14-69[45.1 ±19.6]years) treated in our department of the First Hospital of Shijiazhuang from January 2003 to July 2015 .All patients underwent ultrasound , CT and MRI examination , as well as surgical resection . Results Among these 25 CN patients , there were 15 cases of partial nephrectomy , 5 ca-ses of nephrectomy , 2 cases of retroperitoneal laparoscopic cyst unroofing , 1 case of laparoscopic enucleation of the tumors with nephron-sparing surgery , 1 case of laparoscopic radical nephroureterectomy , and 1 case of retroperitoneal laparoscopic radical nephrectomy .All the patients were successfully followed up for 6 months to 132 months, 1 patient recurred 6 months after retroperitoneal laparoscopic cyst unroo-fing and underwent open partial nephrectomy .No recurrence and me-tastasis were found in the remaining patients . Conclusion Imaging examination is an important measurement for CN , and intraoperative frozen pathology contributes to pathological diagnosis .Since most CN cases are benign , CN patients with no symptom or small cysts can take follow up survey .The principle of the operation is complete resection of the tumor , and nephron-sparing surgery is the first choice . In addition , regular follow-up is necessary in case of recurrence and malignant potential .

Objective To report the experiertce of management of complicated renal stones by percu taneous nephrolithotripsy (PCNL) with pneumatic and ultrasonic power by ultrasound guidance. MethodsThree hundred and eighty two cases(218 males,164 females,4 74 years) who underwent PCNL by u sing the third generation Swiss LithoClast Master for kidney stones from 2004 to 2007 were retrospectivelyreviewed. Clinical data including operation time,stone free rate and complications were analyzed. ResultsPhaseⅠlithotripsy was performed in 397 sides and delayed phaseⅡlithotripsy in 8 sides. Twenty three casesunderwent simultaneous bilateral PCNL. The operation time ranged from 70 to 190 min,average time was(93±11)min. Nine cases needed blood transfusion. Severe complications did not occur during operations.Stone free rate was 91.8% (372/405). Residual stone fragment was found in 33 cases after delayed phase Ⅱlithotripsy and 14 cases received adjuvant extracorporeal shock wave lithotripsy. One hundred and forty sixcases were followed up for 3 to 24 months and showed no recurrence. Conclusion PCNL with pneumaticand ultrasonic power could be an efficient treatment for complicated kidney stones.

Objective To investigate the clinical manifestations,imaging features,molecular genetic characteristics and possible pathogenic mechanisms of hereditary cerebral small vessel disease (CSVD) caused by heterozygous mutation of HtrA serine protease-1 (HTRA1) gene.Methods The clinical data of a Chinese Han family with CSVD carrying a heterozygous mutation of HTRA 1 gene,which came from the Department of Neurology,Henan Provincial People's Hospital in March 2018,were analyzed retrospectively.The clinical and radiographic features were summarized.Several high-throughput whole exon high-throughput sequencing was used to capture the mutation sites and the Sanger sequencing was used to validate the results.The family diagram was drawn and the 3D model construction and mutation function prediction were performed using silico tools.The relevant literature was reviewed and the pathogenesis was explored.Results The pedigree map showed that the family had an autosomal dominant inheritance pattern.Three generations of the family were investigated,and three family members in the same generation suffered from the disease.The first symptom of the proband was diplopia at the age of 39,accompanied by recurrent stroke,cognitive impairment and mood disorders,without alopecia.Head magnetic resonance imaging revealed bilateral diffuse,symmetric lesions,multiple lacunar infarcts,perivascular space,and microbleeds.The elder sister of the proband developed symptoms of left limb weakness at the age of 46,whose other clinical and imaging features were similar to those of the proband.The proband's mother died at the age of 59 due to repeated strokes.Whole exon sequencing indicated heterozygous missense mutation at c.821G＞A locus of HTRA1 gene in the proband and her 4th elder sibling,which was a new pathogenic mutation after consulting several mutation sites of databases.Function prediction suggested pathogenicity.Conclusions The heterozygous mutation of c.821G＞A in HTRA1 gene may lead to autosomal dominant CVSD.This genetic type should be given clinical attention.

Objective:To investigate the effectiveness of an enhanced CT automatic recognition system based on Faster R-CNN for pancreatic cancer and its clinical value.Methods:In this study, 4 024 enhanced CT imaging sequences of 315 patients with pancreatic cancer from January 2013 to May 2016 at the Affiliated Hospital of Qingdao University were collected retrospectively, and 2 614 imaging sequences were input into the faster R-CNN system as training dataset to create an automatic image recognition model, which was then validated by reading 1 410 enhanced CT images of 135 cases of pancreatic cancer.In order to identify its effectiveness, 3 750 CT images of 150 patients with pancreatic lesions were read and a followed-up was carried out.The accuracy and recall rate in detecting nodules were recorded and regression curves were generated.In addition, the accuracy, sensitivity and specificity of Faster R-CNN diagnosis were analyzed, the ROC curves were generated and the area under the curves were calculated.Results:Based on the enhanced CT images of 135 cases, the area under the ROC curve was 0.927 calculated by Faster R-CNN. The accuracy, specificity and sensitivity were 0.902, 0.913 and 0.801 respectively.After the data of 150 patients with pancreatic cancer were verified, 893 CT images showed positive and 2 857 negative.Ninety-eight patients with pancreatic cancer were diagnosed by Faster R-CNN.After the follow-up, it was found that 53 cases were post-operatively proved to be pancreatic ductal carcinoma, 21 cases of pancreatic cystadenocarcinoma, 12 cases of pancreatic cystadenoma, 5 cases of pancreatic cyst, and 7 cases were untreated.During 5 to 17 months after operation, 6 patients died of abdominal tumor infiltration, liver and lung metastasis.Of the 52 patients who were diagnosed negative by Faster R-CNN, 9 were post-operatively proved to be pancreatic ductal carcinoma.Conclusion:Faster R-CNN system has clinical value in helping imaging physicians to diagnose pancreatic cancer.

Objective:To compare the difference in clinical efficacy between a free wrist crease flap pedicled with superficial palmar branch of the radial artery flap (SPBRAF) and a traditional free toe flap (TFTF) in reconstruction of hand soft tissue defects, and to provide reference for the treatment of small-to medium-sized hand soft tissue defects.Methods:Data of 37 patients who received hand surgery in Department of Hand Surgery, No.971 Hospital of the PLA Navy from December 2016 to December 2019 for small-to medium-sized hand soft tissue defects were retrospectively studied. Among the 37 patients, there were 32 males and 5 females, aged between 18 and 65 years old, with 41.5 years old in average. According to the reconstructive surgical procedure, patients were divided into SPBRAF group (22 cases) and TFTF group (15 cases). Regular follow-ups were conducted after surgery. The difference in curative effect at the last follow-up between the 2 groups was evaluated by the comparison of data acquired in follow-up. SPSS 25.0 was used to analyse the data statistically. The evaluation indicators included flap survival, long-term recovery of flap, recovery effect at donor site, total active movement(TAM) of the affected digit, time of hospital stay and the time return to work. P<0.05 was considered a statistically significant. Results:All free flaps survived. All patients were entered 6-18 (mean, 10) months of postoperative follow-up to comprehensively evaluate the therapeutic effect. According to the Evaluation Trial Standards of Upper Limb Partial Function of Hand Surgery of Chinese Medical Association, in the SPBRAF group, 20 flaps were found in excellent, and 2 in good; in the TFTF group, 14 flaps were found in excellent, 1 in good. There was no statistical difference between the 2 groups( P>0.05). The colour, texture and thickness of flaps between the 2 groups were either in excellent or good. There was no statistical difference between the 2 groups( P>0.05). TPD in the TFTF group (5-6 mm) was better than that in SPBRAF group (6-7 mm) with statistical difference between the 2 groups ( P<0.05). Texture at donor sites between the 2 groups was either in excellent or good ( P>0.05). In terms of appearance, sensation and recovery time of donor site, it was found that the SPBRAF group(mean, 6 weeks) was significantly better than those in the TFTF group(mean, 8 weeks) and there was statistical difference between the 2 groups ( P<0.05). In terms of recovery of TAM in single-digit, excellent or good were shown in both groups and there was no statistical difference between the 2 groups ( P>0.05). In terms of hospitalisation and time for return to work, the SPBRAF group(mean, 8 days and 17 weeks) was significantly better than that of TFTF group(mean, 12 days and 24 weeks), and there was statistical difference between the 2 groups ( P<0.05). Conclusion:SPBRAF has an ideal effect on reconstruction of small-to medium-sized hand soft tissue defects in hand. Although the flap is still inferior in sensation and appearance compared with the TFTF, the advantages in terms of donor site recovery, patient satisfaction of the donor site and reduced time of hospitalisation and return to original work are more obvious. SPBRAF provides a good complement to surgical procedures reconstructing a digit defect.

Objective:To investigate the effectiveness of an enhanced CT automatic recognition system based on Faster R-CNN for pancreatic cancer and its clinical value.Methods:In this study, 4 024 enhanced CT imaging sequences of 315 patients with pancreatic cancer from January 2013 to May 2016 at the Affiliated Hospital of Qingdao University were collected retrospectively, and 2 614 imaging sequences were input into the faster R-CNN system as training dataset to create an automatic image recognition model, which was then validated by reading 1 410 enhanced CT images of 135 cases of pancreatic cancer.In order to identify its effectiveness, 3 750 CT images of 150 patients with pancreatic lesions were read and a followed-up was carried out.The accuracy and recall rate in detecting nodules were recorded and regression curves were generated.In addition, the accuracy, sensitivity and specificity of Faster R-CNN diagnosis were analyzed, the ROC curves were generated and the area under the curves were calculated.Results:Based on the enhanced CT images of 135 cases, the area under the ROC curve was 0.927 calculated by Faster R-CNN. The accuracy, specificity and sensitivity were 0.902, 0.913 and 0.801 respectively.After the data of 150 patients with pancreatic cancer were verified, 893 CT images showed positive and 2 857 negative.Ninety-eight patients with pancreatic cancer were diagnosed by Faster R-CNN.After the follow-up, it was found that 53 cases were post-operatively proved to be pancreatic ductal carcinoma, 21 cases of pancreatic cystadenocarcinoma, 12 cases of pancreatic cystadenoma, 5 cases of pancreatic cyst, and 7 cases were untreated.During 5 to 17 months after operation, 6 patients died of abdominal tumor infiltration, liver and lung metastasis.Of the 52 patients who were diagnosed negative by Faster R-CNN, 9 were post-operatively proved to be pancreatic ductal carcinoma.Conclusion:Faster R-CNN system has clinical value in helping imaging physicians to diagnose pancreatic cancer.

Objective:To investigate the role of hepatitis B virus X protein (HBx) in glomerular podocyte immune disorder and its regulatory mechanism.Methods:Fourteen 6-week-old male hepatitis B virus (HBV) transgenic (HBV-Tg) mice were selected, and age-matched wild type (WT) mice were as controls. They were fed to different weeks, and 24 h urinary protein, blood biochemistry, renal pathology and podocyte changes under electron microscope were detected. The expression of HBx and the infiltration of immune cells in kidney tissue of HBV-Tg mice were observed by immunohistochemistry. Human podocyte cell line was transfected with pcDNA3.1/myc-HBx plasmid, and the localization of HBx and Nephrin in podocytes was detected by immunofluorescence. The expression of major histocompatibility complex Ⅱ (MHC-Ⅱ) and co- stimulatory molecule CD40 on the cell surface was detected by flow cytometry. The contents of multiple cytokines in cell culture supernatants were determined by enzyme-linked immunosorbent assay. Transcriptome sequencing (RNA-seq) was used to screen the downstream related genes regulated by HBx, and real-time quantitative PCR was used to verify their expressions. After overexpression or silencing of Notch1 gene with overexpressed plasmids or short hairpin RNA (shRNA) in podocytes, the effects on the expression of immune molecules and cytokines secretion was observed. The Notch receptor inhibitor N-[N-(3, 5-difluorophenyl-l- alanyl)]-(s)-phenylglycine tert-butyl ester (DAPT) was used to block Notch1 signaling pathway in HBV-Tg mice, and then blood biochemistry, renal pathological changes and infiltration of immune cells in kidney tissue were observed. Results:Twenty-four-hour urine protein, serum creatinine and urea nitrogen levels were markedly increased (all P<0.05) and renal pathological injury was significantly aggravated in HBV-Tg mice than those in WT mice. Also, HBx was up-regulated and immune cells infiltrated in the glomerulus of HBV-Tg mice. After transfection with HBx in podocytes, the expression of MHC-Ⅱ and CD40 on the cellular surface was up-regulated (all P<0.05), the contents of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor -α (TNF-α) and interleukin (IL)-1β in the supernatants were increased (all P<0.05), and the secretion of IL-4 and interferon γ (IFN-γ) was unbalanced. RNA-seq screened downstream genes of HBx, such as Notch1, PLA2R, TLR4, etc; and further confirmed that HBx could promote the up-regulation of Notch1 mRNA and protein (all P<0.05). After over-expression of Notch1 gene, HBx-induced expression of MHC-Ⅱ and CD40 on the cellular surface was significantly up-regulated (all P<0.05), and the contents of MCP-1, TNF-α and IL-1β in the supernatants were obviously increased (all P<0.05), and the imbalance of IL-4/IFN-γ was further aggravated. After Notch1 gene silencing, the above results showed the opposite changes. In vivo, the results indicated that serum creatinine levels were obviously decreased (all P<0.05), renal pathological injury and immune cell infiltration were significantly alleviated in HBV-Tg+DAPT group than those in HBV-Tg+DMSO group. Conclusions:HBx protein can promote the up-regulation of Notch1 signaling pathway in podocytes. And Notch1 signaling pathway promotes the expression of immune molecules on the surface of podocytes and regulates the imbalance of cytokines, then causes glomerular injury and dysfunction of immune microenvironment.

African swine fever(ASF)is a virulent,hemorrhagic disease of swine caused by African swine fever virus(ASFV),which seriously affects the healthy development of Chinese pig indus-try.The genome of ASFV is large and encodes more than 165 proteins.Among them,the p54 pro-tein is encoded by the E183L gene,which has various functions such as participating in viral as-sembly,inducing apoptosis and inducing immune response.The conventional Chinese vaccine(C-strain)is a safe and effective attenuated vaccine developed by Chinese scientists.It can efficiently protect against attacks from various genotypes of classical swine fever virus(CSFV).The aim of this study was to investigate whether C-strain can express ASFV p54 protein serve as a delivery vector for ASF genetically engineered vaccines.An infectious clone of pHCLV-p54 was constructed by homologous recombination,the recombinant virus rHCLV-p54 was rescued by transfecting it into SK6 cells by blind passaging.Its genetic stability and growth curve were determined in vitro,while rabbits were immunized to evaluate its immunity effect.The results showed that the E183L gene remained genetically stable in the recombinant virus,indicating that the E183L gene could be stably inherited in recombinant viruses,but the inserted exogenous gene affected the replication of the C-strain.The results of the rabbit immunization test showed that the recombinant virus rHCLV-p54 was able to induce ASFV-specific antibodies.The above results indicated that we have successfully constructed a recombinant C-strain that stably expresses the ASFV p54 protein.In summary,the recombinant virus rHCLV-p54 has a good immunogenicity and is warranted for fur-ther evaluation as a vaccine candidate.