Stomach exuberance and spleen deficiency are common pathogenesis of many metabolic diseases. Through analyzing the pathogenesis of stomach exuberance and spleen deficiency， it is believed that its essence is stomach heat and spleen deficiency. Stomach heat includes gastrointestinal heat， spleen and stomach damp-heat， and spleen deficiency is divided into deficiency of spleen yin， deficiency of spleen qi ， and deficiency of spleen yang. It is suggested that the metabolic diseases of stomach-exuberance and spleen-deficiency syndrome can be divided into three categories，i.e. stomach-heat and spleen yin-deficiency， stomach-heat and spleen qi-deficiency， and stomach-heat and spleen yang-deficiency， and the main treatment methods are clearing and draining heat， nourishing yin and moistening intestine， clearing dampness and heat， strengthening spleen and qi， clearing dampness and heat， strengthening spleen and warming yang， respectively， with prescriptions as Maziren Pills （麻子仁丸）， Qinlian Pingwei Powder （芩连平胃散）， and Jiawei Lianli Decoction （加味连理汤） accordingly.

Objective:To investigate the current status of pertussis laboratory diagnosis and confirmed pertussis cases reporting in Shaanxi Province, and evaluate the quality of case reports.Methods:The information of confirmed pertussis cases reported in Shaanxi Province from January to July 2022 was collected through the China Disease Control and Prevention Information System. The laboratory diagnostic methods and pertussis vaccine immunization history of confirmed cases were investigated, and the descriptive epidemiological method was used for statistical description.Results:Of the 164 confirmed cases of pertussis reported from January to July 2022, two were not tested in the laboratory and 162 were tested in the laboratory. The proportions of different detection methods were 1.85% (3/162) of isolation and culture, 31.48% (51/162) of serum antibody IgG, 14.20% (23/162) of serum antibody IgM, 49.38% (80/162) of serum antibody PCR and 3.09% (5/162) of serum antibody IgM+ PCR. Among the 79 serological positive cases, 12 cases (15.19%)had no history of pertussis immunization, and 15 cases (18.99%), 11 cases (13.92%) and 41 cases (51.90%) had the time interval from vaccination to detection of <1 year, 1-3 years and >3 years, respectively. Based on the analysis of laboratory testing methods and vaccination history, 38 cases were misdiagnosed/misreported among 164 cases, with a misdiagnosis or misreported rate of 23.17%. There were 17, 12 and 9 cases of misdiagnosis/misreport in 0-2 years old group, 3-6 years old group and≥7 years old group, and the misdiagnosis or misreported rates were 27.42%(17/62), 26.67%(12/45) and 15.79%(9/57), respectively.Conclusions:The selection of pertussis laboratory testing methods in some medical institutions in Shaanxi Province is incorrect, which leads to a certain proportion of misdiagnosis or misreport, and it is necessary to further strengthen the training and standardization.

Objective:To prepare the whole bladder acellular matrix (BAM) using the self-designed perfusion decellularization system, and evaluate the feasibility of constructing the tissue engineering bladder with the adipose-derived stem cells (ADSCs).Methods:This study was conducted from October 2020 to April 2021. The self-designed perfusion decellularization system was used, and four different decellularization protocols (group A, group B, group C and group D) were formulated, according to the flow direction of the perfusate and the action time of different decellularization solutions. Among them, the urethral orifice of the bladder tissue was used as the outflow tract of the perfusion fluid in groups A and B. The top of the bladder was cut off and used as the outflow tract of the perfusion fluid in groups C and D. In groups A and C, 1% Triton X-100 was treated for 6 h, and 1% sodium dodecyl sulfate (SDS) was treated for 2 h. In groups B and D, 1% Triton X-100 was treated for 7 h, and 1% sodium dodecyl sulfate (SDS) was treated for 1 h. In addition, the tissue in the normal bladder group was directly obtained from the natural bladder tissue, which did not require perfusion, cryopreservation and thawing. The fast and efficient decellularization protocol was screened out through HE, DAPI, Masson trichrome and Alcian Blue staining and quantitative analyses to prepare the whole bladder scaffold. The prepared BAM was used as the scaffold material, and the ADSCs were used as the seeding cells to construct the tissue engineering bladder. HE and DAPI staining were used to observe the distribution of ADSCs on the BAM.Results:HE and DAPI staining showed that there was no obvious nuclear residue in the group C. Masson trichrome and Alcian Blue staining showed that the collagen structure and glycosaminoglycan were well preserved in the group C. There was no significant difference in bladder wall thickness between the group C and the normal bladder group [(975.44±158.62)μm vs.(1 064.49±168.52)μm, P > 0.05]. The DNA content in the group C [(43.59 ±4.59) ng/mg] was lower than that in the normal bladder group, group A, group B and group D [(532.50±26.69), (135.17±6.99), (182.49±13.69) and(84.00±4.38)ng/mg], and the difference was statistically significant ( P<0.05). The collagen content [(10.98 ± 0.29)μg/mg] and glycosaminoglycan content [(2.30±0.18)μg/mg] in group C were not significantly different with those in the normal bladder group [(11.69±0.49) and (2.36±0.09)μg/mg, P>0.05]. Scanning electron microscopy showed that a large number of pore structures could be observed on the surface of the prepared BAM in groups A-D and were facilitated to cell adhesion. The isolated and cultured ADSCs were identified by flow cytometry to confirm the positive expression of CD90 and CD29, and the negative expression of CD45 and CD106. Live/dead staining and CCK-8 detection confirmed that the prepared BAM in the group C had no cytotoxicity. HE and DAPI staining showed that a large number of ADSCs were distributed on the surface and inside of the tissue engineering bladder. Conclusions:The whole bladder shape BAM prepared by the self-designed perfusion decellularization system could be used as the scaffold material for bladder tissue engineering, and the constructed tissue engineering bladder could be used for bladder repair and reconstruction.

Objective:This study aimed to investigate the physical properties, biocompatibility, and 3D printing performance of a novel hybrid bioink composed of gelatin methacrylated (GelMA) and chitin nanocrystal (ChiNC).Methods:The study was conducted from May 2021 to December 2022, four different bioinks were prepared by adding varying amounts of ChiNC to GelMA bioink. The GelMA concentration in all four bioinks was 100 mg/ml, while the ChiNC concentrations were 0 mg/ml (no ChiNC added), 5 mg/ml, 10 mg/ml, and 20 mg/ml, respectively, named as GC0, GC5, GC10, and GC20 bioinks. The cross-sectional morphology of the hydrogels formed after photocuring the four bioinks was observed using scanning electron microscopy, and the porosity was calculated. Weighing the hydrogels before and after swelling, and then calculate the equilibrium swelling rate. HUVECs were seeded on the surfaces of the hydrogels prepared from the four bioinks and cultured in medium. Cell proliferation was assessed using CCK-8 assays at 1d, 3d, and 7d to compare the proliferation rates of cells on the four hydrogels. HUVECs were added to the four bioinks, and grid-like scaffolds were printed and cultured in medium. Live-Dead staining was performed at 1d and 7d to observe cell viability. Compare the printing effect of bioinks by observing its forming continuous threads properties during extrusion. Finally, tissue-engineered bladder patches simulating the mucosal layer, submucosal layer, and muscular layer anatomical structures of the bladder wall were 3D bioprinted using the optimized bioink composition, and the stability and fidelity of the printed structures were observed to further validate the feasibility of printing multi-layered complex structures with the bioink.Results:Scanning electron microscopy revealed that the porosity of the GC0, GC5, GC10, and GC20 hydrogels were (51.43±6.23)%, (51.85±6.47)%, (50.55±4.59)%, and (42.49±2.20)%, respectively. The differences in porosity between the GC0 group and the other three groups were not statistically significant ( P=0.9994, P=0.9948, P=0.1200). The equilibrium swelling ratio of the other three groups [(8.81±0.41), (7.95±0.19), (7.71±0.14)] was significantly lower than that of the GC0 group (9.37 ± 0.49), and the differences were statistically significant ( P=0.0457, P<0.01, P<0.01). CCK-8 assay showed no significant difference in absorbance value between the GC10 group (0.360±0.009) and the GC0 group (0.357±0.007), GC5 group (0.350±0.012), and GC20 group (0.345±0.018) on the first day ( P=0.9332, P=0.5464, P=0.4937). However, on the third day, the absorbance value of the GC10 group (0.755±0.012) was significantly higher than that of the GC0 group (0.634±0.010), GC5 group (0.704±0.009), and GC20 group (0.653±0.015) ( P<0.01, P=0.0033, P=0.0002). On the seventh day, the absorbance value of the GC10 group (1.001±0.031) was significantly higher than that of the GC0 group (0.846±0.026), GC5 group (0.930±0.043), and GC20 group (0.841±0.024)( P=0.0012, P=0.1390, P=0.0010). The addition of human umbilical vein endothelial cells (HUVECs) into the four groups of hydrogels enabled the printing of grid-like scaffolds, and Live-Dead staining was performed on day 1 and day 7. The cell viability of HUVECs in the four groups on day 1 was (90.13±1.63)%, (90.6±2.45)%, (92.58±2.15)%, and (91.40±3.17)%, respectively. There were no statistically significant differences between the GC0 group and the other three groups ( P=0.9869, P=0.3093, P=0.8008). On day 7, the cell viability was (89.97±3.10)%, (92.18±2.21)%, (92.05±2.25)%, and (90.12±1.97)% for the four groups, respectively. There were no statistically significant differences between the GC0 group and the other three groups ( P=0.3965, P=0.4511, P=0.9995). Bioink extrusion test showed that the GC0 hydrogel could be extruded continuously and form threads at temperatures between 24℃ and 25℃, while the GC10 hydrogel could be extruded continuously and form threads at temperatures between 24℃ and 27℃. Printing tissue engineered bladder patches simulating the anatomical structure of the bladder mucosal layer, submucosal layer, and muscular layer using GC10 bioink, and the printed patches were stable, without collapse, and had high fidelity. Conclusions:Adding ChiNC to GelMA promotes cell adhesion, proliferation, and expands the printing window of GelMA bioink. The biocompatibility of the mixed bioink prepared by adding 10 mg/ml ChiNC in GelMA is good, capable of printing tissue-engineered bladder patches that mimic the anatomical structure of natural bladder walls.

Objective:To investigate the predictive value of MRI radiologic extranodal extension (rENE) for distant metastasis of prostate cancer (PCa).Methods:The data of 107 patients of initial visit with clinically diagnosed N1 PCa who underwent MRI and 68Ga-prostate specific membrane antigen (PSMA) PET/CT examinations were retrospectively analyzed at Xijing Hospital, Air Force Medical University from January 2017 to April 2022. The rENE was evaluated with MRI. According to the results of 68Ga-PSMA PET/CT, the patients were divided into the distant metastasis group (group M1, 87 cases) and the non-distant metastasis group (group M0, 20 cases). Independent sample t test, Mann-Whitney U test or χ 2 test were used to compare the differences in clinical indicators and rENE between the two groups. The multivariate logistic regression analysis was used to screen the independent risk factors affecting distant metastasis. The receiver operating characteristic (ROC) curve was used to evaluate the predictive efficacy of independent risk factors for PCa distant metastasis. Results:In group M1, 72 cases (82.8%) were rENE positive and 15 cases (17.2%) were rENE negative, and in group M0, 7 cases (35.0%) were rENE positive and 13 cases (65.0%) were rENE negative, and there was a statistically significant difference in rENE between the two groups (χ 2=19.20, P<0.001). There were significant differences in total prostate specific antigen level, International Society of Urological Pathology grade and T stage between the group M1 and the group M0 ( P<0.05). Multivariate logistic regression analysis showed that rENE (OR=6.248, 95%CI 1.807-21.600, P=0.004) was an independent risk factor for distant metastasis of PCa, and the area under the ROC curve of rENE in the diagnosis of distant metastasis of PCa was 0.739 (95%CI 0.607-0.871), the sensitivity was 82.8%, and the specificity was 65.0%. Conclusion:rENE is an independent predictor of distant metastasis of PCa, which has a high efficacy. Compared with patients with rENE negative, PCa patients with rENE positive have a higher degree of invasion and are more likely to have distant metastasis.

Objective:To investigate the significance of clinical factors combined with transvaginal ultrasound and contrast-enhanced ultrasound(CEUS) in guiding the choice of treatment plan for cesarean scar pregnancy(CSP).Methods:The clinical and transvaginal ultrasound and CEUS data of 120 patients with CSP from January 2016 to June 2021 in the First People′s Hospital of Foshan were retrospectively analyzed, and they were divided into ultrasound-guided curettage/ hysteroscopic group (Group A, 91 cases) and laparoscopic group (Group B, 29 cases) according to treatment option, and the differences in clinical and ultrasound factors between the two groups were compared, and to determine the relevant clinical and ultrasound indicators for the choice of treatment option.Results:There were statistical differences between the 2 groups in comparison of whether the gestational sac/mass protruded toward the plasma membrane, gestational sac/mass diameter, the main blood supply site of the gestational sac/mass, the site of the chorion/early placenta and scar thickness (all P<0.05). Logistic regression analysis indicated that CEUS showing major blood supply site of the gestational sac/mass ( OR=6.029, P=0.003) and uterine scar thickness ( OR=12.998, P=0.002) were independent risk factors for minimally invasive surgery for CSP. Conclusions:Ultrasound combined with clinical factors have a certain value in the selection of treatment options for CPS, and the thickness of the uterine scar and the main blood supply site of the gestational sac/mass showed in CEUS may be key factors affecting the minimally invasive surgical treatment of CSP.

For the damage and loss of tissues and organs caused by urinary system diseases, the current clinical treatment methods have limitations. Tissue engineering provides a therapeutic method that can replace or regenerate damaged tissues and organs through the research of cells, biological scaffolds and biologically related molecules. As an emerging manufacturing technology, three-dimensional (3D) bioprinting technology can accurately control the biological materials carrying cells, which further promotes the development of tissue engineering. This article reviews the research progress and application of 3D bioprinting technology in tissue engineering of kidney, ureter, bladder, and urethra. Finally, the main current challenges and future prospects are discussed.
Bioprinting ; Regeneration ; Technology ; Tissue Engineering/methods*

Objective:To investigate the effect of methyltransferase like 14 (METTL14) on the proliferation and invasion of breast cancer (BC) cells by regulating cyclin L2 (Cyclin L2, CCNL2) through m6A modification.Methods:Cancer tissues and paracancerous tissues of BC patients in Yantaishan hospital were collected from Aug. 2018 to Feb. 2020. The expression levels of m6A, METTL14 and CCNL2 in tissues were detected by high performance liquid chromatography/mass spectrometry (HPLC/MS) and qRT-PCR. Dual-luciferase reporter assay, qRT-PCR, and western blot were used to verify the regulatory relationship between METTL14 and CCNL2. RIP experiments verified the regulatory relationship between YTH domain-containing family protein (YTHDF2) and CCNL2. Cell viability was detected by MTT method, and cell invasion ability was detected by Transwell.Results:Compared with normal cells (0.24±0.02) and tissues (0.18±0.02) , BC cells MCF-10A (0.47±0.03, t=11.05, P<0.001) and HS-578T (0.41±0.03, t=8.17, P=0.001) and BC tissues (0.39±0.02, t=12.86, P<0.001) m6A level increased. Compared with normal tissues (1.00±0.26) (0.84±0.07) , METTL14 mRNA (1.57±0.28, t=13.50, P<0.001) and protein levels (1.66±0.11, t=10.89, P<0.001) in BC tissues were significantly increased high. Compared with the control group (100.00±10.11) (1.00±0.12) , the BC cell invasion ability (54.15±6.21, t=6.69, P=0.003) and activity (0.64±0.06, t=4.65, P=0.010) were weakened. Compared with the control group (100±11.05) (1±0.13) , the BC cell invasion ability (175.31±13.45, t=7.49, P=0.002) and activity (2.16±0.16, t=9.75, P=0.002) in the METTL14 overexpression group were enhanced, and the effects of METTL14 on cell invasion (137.41±12.64, t=3.56, P=0.024) and activity (1.64±0.15, t=5.59, P=0.005) were partially reversed after m6A inhibitor treatment change. Compared with normal tissues, CCNL2 expression was down-regulated in BC tissues, and the interaction between CCNL2 and METTL14 was confirmed. Compared with the control group (1.00±0.1) (0.64±0.05) , knockdown of METTL14 could make CCNL2 mRNA (1.67±0.05) . 0.13, t=7.08, P=0.002) and protein (1.09±0.09, t=7.57, P=0.002) were up-regulated. METTL14 knockout enhanced the stability of CCNL2 mRNA through a YTHDF2-dependent pathway, compared with sh-METTL14 group (50.47±5.16) (0.52±0.05) , BC cell invasion ability of sh-METTL14+sh-CCNL2 group (71.69±6.41, t=4.47, P=0.011) and activity (0.64±0.05, t=2.94, P=0.042) were improved. Conclusion:METTL14 inhibits the expression of CCNL2 through m6A modification to enhance the invasion and activity of BC cells.

Objective:To explore the expectations and values of patients with malocclusion on participation in shared decision-making of orthognathic surgical protocols, and to provide references for further development of clinical shared decision-making models.Methods:Based on the expected value theory and descriptive qualitative research methods, using purposive sampling, 13 patients with malocclusion in the Ninth People's Hospital of Shanghai Jiao tong University School of Medicine from May to August in 2021 were selected for semi-structured interviews. The interview data were sorted, classified and refined by traditional content analysis.Results:Two themes were extracted: patients' ability beliefs about their ability to participate in shared decision making for orthognathic surgery(decision support ability, psychological coping ability and environmental adaptability), and task values for shared decision making for orthognathic surgery(interest value, acquisition value).Conclusions:Low level of patients′ ability beliefs in shared decision-making, active physician guidance facilitates patient participation, but the depth of patient participation is influenced by factors such as information support, cultural climate, and physical space for shared decision making. It is suggested that the magnetic role of physicians should be actively played, the shared decision-making team should be strengthened, at the same time, hospital manager should enhance shared decision making propaganda to increase the acceptance and participation of patients in shared decision making so as to improve the quality of shared decision-making.

Objective:To investigate the effect of tissue engineered bladder patch constructed by double-layer silk scaffold and adipose-derived stem cells (ADSCs) in the repair and reconstruction of bladder.Methods:This study was conducted from May 2020 to March 2021. The silk fibroin (SF) aqueous solution was obtained from silkworm cocoons, and a double-layer silk scaffold composed of silk fibroin film and silk fibroin sponge was further prepared. The rat ADSCs were isolated, cultured, and the ADSCs surface markers (CD29, CD90, CD45, CD106) were identified by flow cytometry. The ADSCs were planted on a double-layer silk scaffold to construct a tissue-engineered bladder patch. Thirty-six male SD rats were randomly divided into three groups: tissue engineered bladder patch group (SF-ADSCs group, n=15), double-layer silk scaffold group (SF group, n=15), control group ( n=6). The tissue engineered bladder patch (SF-ADSCs group) and double-layer silk scaffold (SF group) were wrapped on the omentum to promote vascularization. The vascularization was evaluated by HE and immunofluorescence staining. The wrapped tissue engineered bladder patch and double-layer silk scaffold were used to repair the defective bladder. In the control group (six rats), the incision was closed immediately after the bladder tissue fully exposed. At 4 weeks and 12 weeks after operation, the general morphology of bladder tissue and cystography were performed to evaluate the recovery of bladder morphology. After the graft was harvested, HE and Masson's trichrome staining and immunofluorescence staining were used to observe the regeneration of bladder wall tissue. Urodynamics was used to assess the recovery of bladder function at 12 weeks after operation. Results:The flow cytometry results confirmed that the isolated cells positively expressed CD29 and CD90, and there was no significant expression of CD45 and CD106. Gross observation and scanning electron microscope confirmed that the preparation of double-layer silk scaffold not only had a pore structure that was conducive to cell planting, but also had good toughness and was facilitated to surgical suture. The number (43.50±2.66) and area (0.73±0.03)% of vascular-like structures in the SF-ADSCs group after the omentum encapsulation was significantly higher than that in the SF group [(24.50±3.51), (0.55±0.05)%], and the difference was statistically significant ( P<0.05). At 4 weeks after bladder repair, the histological staining of the grafts in the SF-ADSCs and SF groups showed a large number of degraded fragments of double-layer silk scaffold. At 12 weeks, the morphology of the graft in the SF-ADSCs group showed uniform bladder morphology, which was similar to that of normal bladder tissue. Immunofluorescence staining showed that the continuous urothelial layer, abundant smooth muscle tissue, vascular structure and regenerated neurons could be observed in the SF-ADSCs group. Urodynamic test showed that the bladder maximum volume (0.74±0.03)ml and compliance (16.68±0.44)μl/cm H 2O in the SF-ADSCs group, which were better than that in the SF group [(0.47±0.05)ml, (14.89±0.37)μl/cm H 2O], but lower than that in the control group [(1.12±0.08)ml, (19.34±0.45)μl/cm H 2O], and the difference was statistically significant ( P<0.05). Conclusions:The tissue engineered bladder patch constructed with double-layer silk scaffolds and ADSCs could promote the morphological repair of bladder tissue, the regeneration of bladder wall structure and the recovery of bladder physiological function.