The objective of present study was to develop a simple and reliable voiding spot assay (VSA) system to evaluate the lower urinary tract function of mice, and to establish it as a standardized protocol. Ultraviolet (UV) light was used to screen out the filter paper without autofluorescence and with optimal urine diffusion properties. Next, the appropriate wavelength of UV was determined based on the quality of the photographic image of urine spots on the filter paper. To confirm that the urine stain area on the filter paper was correlated with the amount of urine, a volume-area standard curve was constructed. The utility of this VSA system was validated using female wild-type C57BL/6J mice aged 12-13 weeks, and the data generated under identical procedural settings were compared among laboratories. Furthermore, this VSA system was employed to analyze the changes in voiding patterns in mice with urinary tract infections or transportation stress. No. 4 filter paper with a thickness of 0.7 mm was identified as the most suitable material for VSA, exhibiting no autofluorescence and facilitating optimal urine diffusion. The filter paper retained its integrity during the assay, and there was a linear correlation between urine volume and stained area under 365 nm UV light. Utilizing this VSA system, we determined that female wild-type C57BL/6J mice produced approximately 695.8 μL total urine and 5.5 primary voiding spots (PVS) with an average size of 126.4 μL/spot within 4-h period. Over 84% of PVS volumes ranged from 20 to 200 μL. Notably, PVS volumes of mice were similar across different laboratories. Mice with urinary tract infections or transportation stress exhibited significant changes in VSA parameters, including increased voiding frequency, PVS number, and decreased PVS volume. Therefore, this VSA system can be used to evaluate the urinary function of normal mice, as well as those with urinary tract infection or transportation stress.
Mice ; Female ; Animals ; Urodynamics ; Mice, Inbred C57BL ; Urination ; Urinary Bladder ; Urinary Tract Infections

This study aimed to investigate the effect and molecular mechanism of sinomenine on proliferation, apoptosis, metastasis, and combination with inhibitors in human hepatocellular carcinoma HepG2 cells and SK-HEP-1 cells. The effect of sinomenine on the growth ability of HepG2 and SK-HEP-1 cells were investigated by CCK-8 assay, colony formation assay, and BeyoClick~(TM) EdU-488 staining. The effect of sinomenine on DNA damage was detected by immunofluorescence assay, and the effect of sinomenine on apoptosis of human hepatocellular carcinoma cells was clarified by Hoechst 33258 staining and CellEvent~(TM) Cystein-3/7Green ReadyProbes~(TM) reagent assay. Cell invasion assay and 3D tumor cell spheroid invasion assay were performed to investigate the effect of sinomenine on the invasion ability of human hepatocellular carcinoma cells in vitro. The effect of sinomenine on the regulation of protein expression related to the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription 3(STAT3) signaling pathway in HepG2 and SK-HEP-1 cells was examined by Western blot. Molecular docking was used to evaluate the strength of affinity of sinomenine to the target cysteinyl aspartate specific proteinase-3(caspase-3) and STAT3, and combined with CCK-8 assay to detect the changes in cell viability after combination with STAT3 inhibitor JSI-124 in combination with CCK-8 assay. The results showed that sinomenine could significantly reduce the cell viability of human hepatocellular carcinoma cells in a concentration-and time-dependent manner, significantly inhibit the clonogenic ability of human hepatocellular carcinoma cells, and weaken the invasive ability of human hepatocellular carcinoma cells in vitro. In addition, sinomenine could up-regulate the cleaved level of poly ADP-ribose polymerase(PARP), a marker of apoptosis, and down-regulate the protein levels of p-Akt, p-mTOR, and p-STAT3 in human hepatocellular carcinoma cells. Molecular docking results showed that sinomenine had good affinity with the targets caspase-3 and STAT3, and the sensitivity of sinomenine to hepatocellular carcinoma cells was diminished after STAT3 was inhibited. Therefore, sinomenine can inhibit the proliferation and invasion of human hepatocellular carcinoma cells and induce apoptosis, and the mechanism may be attributed to the activation of caspase-3 signaling and inhibition of the Akt/mTOR/STAT3 pathway. This study can provide a new reference for the in-depth research and clinical application of sinomenine and is of great significance to further promote the scientific development and utilization of sinomenine.
Humans ; Carcinoma, Hepatocellular/genetics* ; Proto-Oncogene Proteins c-akt/metabolism* ; Caspase 3/metabolism* ; Liver Neoplasms/genetics* ; Molecular Docking Simulation ; Sincalide/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Hep G2 Cells ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis

This study investigated the effect and mechanism of morin in inducing autophagy and apoptosis in hepatocellular carcinoma cells through the protein kinase B(Akt)/mammalian target of rapamycin(mTOR)/signal transducer and activator of transcription protein 3(STAT3) pathway. Human hepatocellular carcinoma SK-HEP-1 cells were stimulated with different concentrations of morin(0, 50, 100, 125, 200, and 250 μmol·L~(-1)). The effect of morin on the viability of SK-HEP-1 cells was detected by Cell Counting Kit-8(CCK-8). The effect of morin on the proliferation and apoptosis of SK-HEP-1 cells was investigated using colony formation assay, flow cytometry, and BeyoClick~(TM) EdU-488 with different concentrations of morin(0, 125, and 250 μmol·L~(-1)). The changes in the autophagy level of cells treated with morin were examined by transmission electron microscopy and autophagy inhibitors. The impact of morin on the expression levels of proteins related to the Akt/mTOR/STAT3 pathway was verified by Western blot. Compared with the control group, the morin groups showed decreased viability of SK-HEP-1 cells in a time-and concentration-dependent manner, increased number of apoptotic cells, up-regulated expression level of apoptosis marker PARP, up-regulated phosphorylation level of apoptosis-regulating protein H2AX, decreased number of positive cells and the colony formation rate, an upward trend of expression levels of autophagy-related proteins LC3-Ⅱ, Atg5, and Atg7, and decreased phosphorylation levels of Akt, mTOR, and STAT3. These results suggest that morin can promote apoptosis, inhibit proliferation, and induce autophagy in hepatocellular carcinoma cells, and its mechanism of action may be related to the Akt/mTOR/STAT3 pathway.
Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; TOR Serine-Threonine Kinases/metabolism* ; Apoptosis ; Autophagy ; Cell Proliferation ; Cell Line, Tumor ; STAT3 Transcription Factor/metabolism*

Objective:To study the policy framework and core content of assistive technology (AT) services, compare the current status of AT in countries with different levels of development, and discuss the future development of international policies and services of AT. Methods:Based on the policy and theoretical framework of AT of the United Nations (UN) and World Health Organization (WHO), the policy framework and main contents of international AT services were analyzed; the policies, assistive products and service development of AT services in countries with different levels of development worldwide were compared; and the policy and innovative technology development hotspots of international AT were explored. Results:AT service is an important component of rehabilitation services, and the development of AT services is a key step toward achieving the UN Sustainable Development Goal 3, "Ensure healthy lifestyles and promote the well-being of people of all ages". UN and WHO advocated AT services policy. The framework is based on the UN Convention on the Rights of Persons with Disabilities (CRPD) on articles related to AT, and WHO documents of AT and rehabilitation, such as World Report on Disability, the Global Plan of Action on Disability, Rehabilitation in the Health System, and Rehabilitation in the Health System: A Guide to Action, as well as WHA 71.8 on Assistive Technology, Improving Access to Assistive Technology. The core elements of the international AT policy aim to build integrated, people-centered health services, emphasize the integration of AT into the health system and rehabilitation services, establish and develop AT services in six building blocks of WHO health system, thereby improving access to quality AT to achieve universal health coverage. Further in the fields of technology and service innovation, it proposed to establish a 5P model. High-income countries and low- and middle-income countries are facing many difficulties and large differences in AT service policies, assistive product provision, and service coverage. In the future, adoption of the 5P model proposed by WHO for policy and service delivery and technological changes, and the innovation of assistive product development and AT service models will be the hot spots of AT development. Conclusion:The policy on AT at international level is centered on the concept of UN CRPD and is based on prospect of policy documents issued by the UN and WHO on AT services, with the core objective of developing AT services to provide timely, appropriate, and affordable to persons with disabilities, aging, and people needed to improve their health, quality of life and well-being. It proposes to integrate AT into universal health coverage and to deliver AT service through primary health care in order to achieve the UN SDG 3 goal with full coverage of AT. It proposes to integrate AT into rehabilitation services, i.e. leadership and governance, financing, AT professionals, AT delivery, medicine and technology, and health information system monitoring AT services. There are differneces in AT products and AT delivery due to the globel differneces in economic and social development levels and challenges in access to AT services. It is necessary to develop relevant policies, planning and innovations in assistive products and AT services. AT development in the future will focus on the 5P model of AT to implement reforms in the field of product and service delivery as well as technological innovation in order to improve the coverage, availability, accessibility and affordability, integrate AT into the health system within the framework of universal health coverage, achieve full coverage of universal AT services, and enhance the quality of AT and improve consumers' well-being.

Objective:To study the prevalence of people with disability, and the demand, service policies and service system for assistive technology (AT) services in Australia. Methods:Referring to policy recommendation and indicators in World Health Organizaion's Rehabilitation in Health Systems, Australia's assistive technology service policy framework, services development and trend of development were analyzed by the means of content analysis and development research. Results:There are about 4.4 million people with disability in Australia, of which 1.4 million (32%) with severe or profound disabilities. Federal, state and territorial governments have respectively issued legislation, policy and related service project, including the National Disability Strategy, National Disability Agreement, the National Disability Insurance Scheme, National Standards for Disability Services and others assistance programs to support disability services. There are about two million Australians with disabilities received AT services support from the mentioned programme and scheme. Australia provides more than ten thousands kinds of AT appliances per year, covering all kinds of products in WHO's Priority Assistive Products List. Conclusion:Australia has developed a federal National Disability Strategy, AT service policies and standards, and established federal and state AT insurance and financial support programs for people with disabilities based on the unmet needs of AT services, and federal and state networks for AT services have been established, and service coordination mechanisms at federal and state levels have been established through the National Disability Agreement, covering all types of people with demand of AT in Australia. It meets the requirements of the policy recommendations and assessment indicators in the WHO's Rehabilitation in Health Systems policy guidelines involving AT services. The future development areas in AT will focus on the fields of policy development, ICF implementation, service delivery system, upgrading of service quality and standard, and new technology application.

Objective:To evaluate the clinical effect of applying peroneal artery perforator flap with super-draining techniques in repairing skin defects of extremities.Methods:The data of patients with limb wounds admitted to the 990th Hospital of the Joint Logistics Support Force of the People’s Liberation Army from March 2014 to August 2019 were analyzed retrospectively. All were repaired with peroneal artery perforator flaps with super-draining techniques.Cutaneous nerves and perforating vessels were anastomosed according to the routine ratio of 1∶2.The superficial vein and the recipient veinwere matched and marked according to the relative position the diameter and length.Then one to three superficial veins were anastomosed in sequence according to the vein mark number.Upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Hand Surgery Branch of the Chinese Medical Association.Patients with lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hindfoot Score System.All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association.Results:A total of 67 cases were included in this cohort. There were 54 males and 13 females, aged from 21 to 65 years, with an average of 44 years.Eight cases were anastomosed with three superficial veins, 38 cases were anastomosed with two flaps, and 21 cases were anastomosed with one flap.The donor sites were closed directly in 56 cases.Eleven cases received full-thickness skin grafting to cover the donor site.One case showed necrosis of the half flap for the arterial crisis. The wound was repaired by full-thickness skin grafting in further treatment. Three cases had hemorrhage under the free flap, which was healed secondarily after debridement. The remaining were healed well.All patients were followed for 6 to 44 months. There was no obvious swelling and blisters in the early postoperative period.The texture of the flap was the same as the surrounding area.In the later stage, the atrophy and scar were not obvious, without pigmentation. The sensation recovered quickly. The donor site healed without dysfunction.Sixteen patients with upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Branch. Twelve cases were excellent, 3 cases were good, and one case was fair. Fifty-one cases of lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hind Foot Score System. Thirty-eight cases were excellent, 11 cases were good, and two cases were fair. All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association. The two-point distance discrimination was 8-10 mm, with an average of 9.1 mm. The sensory function of the flaps reached S3+ in 48 cases and S3 in 19 cases.Conclusions:Application of super-draining technique in peroneal artery perforator flap surgery can effectively prevent blood congestion and reduce the necrosis risk of the free flap and complication rate. It can improve the survival quality of the flap and facilitate the recovery of the function of the receiving area.

Objective:To evaluate the clinical effect of applying peroneal artery perforator flap with super-draining techniques in repairing skin defects of extremities.Methods:The data of patients with limb wounds admitted to the 990th Hospital of the Joint Logistics Support Force of the People’s Liberation Army from March 2014 to August 2019 were analyzed retrospectively. All were repaired with peroneal artery perforator flaps with super-draining techniques.Cutaneous nerves and perforating vessels were anastomosed according to the routine ratio of 1∶2.The superficial vein and the recipient veinwere matched and marked according to the relative position the diameter and length.Then one to three superficial veins were anastomosed in sequence according to the vein mark number.Upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Hand Surgery Branch of the Chinese Medical Association.Patients with lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hindfoot Score System.All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association.Results:A total of 67 cases were included in this cohort. There were 54 males and 13 females, aged from 21 to 65 years, with an average of 44 years.Eight cases were anastomosed with three superficial veins, 38 cases were anastomosed with two flaps, and 21 cases were anastomosed with one flap.The donor sites were closed directly in 56 cases.Eleven cases received full-thickness skin grafting to cover the donor site.One case showed necrosis of the half flap for the arterial crisis. The wound was repaired by full-thickness skin grafting in further treatment. Three cases had hemorrhage under the free flap, which was healed secondarily after debridement. The remaining were healed well.All patients were followed for 6 to 44 months. There was no obvious swelling and blisters in the early postoperative period.The texture of the flap was the same as the surrounding area.In the later stage, the atrophy and scar were not obvious, without pigmentation. The sensation recovered quickly. The donor site healed without dysfunction.Sixteen patients with upper limb wounds were evaluated by the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Branch. Twelve cases were excellent, 3 cases were good, and one case was fair. Fifty-one cases of lower limb wounds were evaluated by the American Orthopaedic Foot and Ankle Surgery Association Ankle-Hind Foot Score System. Thirty-eight cases were excellent, 11 cases were good, and two cases were fair. All patients were evaluated according to the sensory function evaluation standard of the British Medical Research Association. The two-point distance discrimination was 8-10 mm, with an average of 9.1 mm. The sensory function of the flaps reached S3+ in 48 cases and S3 in 19 cases.Conclusions:Application of super-draining technique in peroneal artery perforator flap surgery can effectively prevent blood congestion and reduce the necrosis risk of the free flap and complication rate. It can improve the survival quality of the flap and facilitate the recovery of the function of the receiving area.

Objective:To explore the efficacy of percutaneous osteoplasty (POP) combined with 131I therapy in patients with bone metastases from differentiated thyroid carcinoma (DTC) and assess the survival. Methods:From Januray 2008 to January 2020, 29 DTC patients with bone metastases (16 males, 13 females, age range: 24-64 years) who received POP combined with 131I therapy in Shanghai Sixth People′s Hospital were retrospectively analyzed. Clinical data and characteristics of patients were analyzed. The efficacy and prognosis were evaluated based on the changes of serum thyroglobulin (Tg) and relieving condition of bone pain after the combined treatment. χ2 test was used to determine the association between clinical characteristics and efficacy, and Kaplan-Meier analysis was used to estimate the overall survival (OS) rate. Results:The biochemical response rate of serum Tg was 68.97% (20/29) after the combined treatment. For the influence on changes of serum Tg, whether or not combined with non-osseous distant metastasis, and cumulative dose of 131I treatment(≥22.2 vs <22.2 GBq) were statistically significant ( χ2 values: 5.448 and 4.371, both P<0.05). The rate of bone pain relief was 65.52%(19/29). Age (≥55 vs <55 years) and the cumulative dose of 131I treatment had statistically influences on bone relief ( χ2 values: 7.486 and 5.154, both P<0.05). The 5-years OS rate of patients was 87.68%, while the 10-years OS rate was 65.76%. Conclusion:POP combined with 131I therapy is effective on relieving the pain, reducing the serum Tg to some extent, and improving the long-term survival of DTC patients with bone metastasis.

The chemical constituents of the aerial parts of Lespedeza cuneata (Dum. Cour.) G. Don were investigated using chromatographic techniques, such as silica gel, reversed phase MPLC and preparative HPLC. Five compounds were isolated and their structures were elucidated by spectroscopic data and physicochemical properties, which were identified as 7-O-glucosyllaburnetin (1), kaempferol-3-O-β-D-galactopyranoside (2), kaempferol-3-O-α-L-rhamnoside (3), vitexin (4), and isovitexin (5). Among those, compound 1 is a new compound, compounds 2-3 were isolated from this plant for the first time. Compounds 1-5 were tested for their anti-ulcerative colitis activity by dual luciferase report gene assay targeting xbp1. Compared with control group, compound 1 showed a certain activity on activating the transcription of xbp1, with its relative activating ratio being 1.80 times.

Objective To evaluate the effect of Weisu granule combined with conventional therapy in treatment of chronic atrophic gastritis (CAG). Methods A total of 99 CAG patients who met the inclusion criteria were randomly divided into control group (49 cases) and observation group (50 cases) according to random number table method. The control group was treated with quadruple therapy, while the observation group was treated with Weisu granule on the basis of the control group. Both groups were treated for 90 days. Interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α) were detected by ELISA. The scores of TCM symptoms between the two groups before and after treatment were compared. Adverse reactions during treatment were observed and recorded. The eradication rate of H.pylori was measured by 14C urea breath test, and the clinical efficacy was evaluated. Results The total effective rate was 92.0% (46/50) in the observation group and 73.5% (36/49) in the control group. There were significant differences between the two groups (χ2=5.975, P=0.015). After treatment, the level of serum IL-6, hs-CRP and TNF-α in the observation group was significantly lower than those in the control group (t=7.112, 7.753, 9.310, respectively, all P<0.01). After treatment, the scores of symptoms such as abdominal distension and pain, emotional disturbance, hypochondrium pain and belching in both groups decreased (P<0.01), and those in the observation group was significantly lower than those in the control group (t=12.892, 12.451, 10.596, respectively, P<0.01). The eradication rate of H.pylori was 90.0% (45/50) in the observation group and 24.5% (12/49) in the control group. There was a significant difference between the two groups (χ2=43.381, P<0.01). Conclusions The Weisu granule combined with quadruple therapy has a definite theraputic effect. It can improve the clinical symptoms of patients, improve the clearance rate of H.pylori, and alleviate inflammation.