OBJECTIVE: To investigate the effects of combined platelet-rich plasma (PRP) and arterial supercharging technique on the survival rate and functional restoration of cross-body region skin flaps in rabbits. METHODS: Twelve healthy 6-month-old New Zealand White rabbits were randomly assigned to 4 groups ( n=3): sham group, PRP group, anastomosis group, and combined treatment group. An axial skin flap with an area of 12 cm×6 cm on the inner side of the hind limbs of all animals were prepared, with the saphenous artery as the main blood supply. Following the ligation of both the proximal and distal ends of the saphenous artery across all groups, the sham group received no further intervention, the PRP group was subjected to PRP injection, the anastomosis group underwent in situ end-to-end anastomosis of the distal saphenous artery, and the combined treatment group received both in situ distal saphenous artery anastomosis and PRP administration. Flap survival was evaluated and recorded on postoperative days 1, 3, and 7, with survival rates calculated accordingly. On day 7, flap tissue samples were harvested for HE staining to assess basal tissue morphology. Additionally, immunohistochemical staining was conducted to detect the expression of α-smooth muscle actin (α-SMA), vascular endothelial growth factor (VEGF), and CD31 in the flap tissues. RESULTS: At postoperative day 1, no significant difference in flap survival rates were observed among the 4 groups ( P>0.05). At day 3, the PRP group showed no significant difference compared to the sham group ( P>0.05); however, both the anastomosis and combined treatment groups exhibited significantly higher survival rates than the sham group ( P<0.05), the combined treatment group further demonstrated superior survival rates compared to both the PRP and anastomosis groups ( P<0.05). At day 7, the combined treatment group maintained significantly higher survival rates than all other groups ( P<0.05), while both the PRP and anastomosis groups exceeded the sham group ( P<0.05). HE staining at day 7 revealed persistent inflammatory cell infiltration, sheet-like erythrocyte deposition, and disordered collagen fibers in the sham group. The PRP group showed nascent microvessel formation and early collagen reorganization, whereas the anastomosis group displayed mature microvasculature with resolved interstitial edema. The combined treatment group exhibited differentiated microvessels with densely packed collagen bundles. Immunohistochemical analysis at day 7 demonstrated significantly larger relative area percentages of α-SMA, VEGF, and CD31 positive cells in the combined treatment group compared to all other groups ( P<0.05). Both the PRP and anastomosis groups also showed significantly higher values than the sham group ( P<0.05). CONCLUSION The combination of PRP and arterial supercharging techniques significantly enhances flap healing, potentially through mechanisms involving augmented angiogenesis and improved blood supply.
Animals ; Rabbits ; Platelet-Rich Plasma ; Surgical Flaps/blood supply* ; Graft Survival ; Anastomosis, Surgical/methods* ; Ischemia/surgery* ; Arteries/surgery* ; Skin/blood supply* ; Vascular Endothelial Growth Factor A/metabolism* ; Male ; Skin Transplantation/methods*

Objective To investigate the effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its association with the regulation of the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear translocation. Methods A total of 32 male Sprague-Dawley rats were randomly divided into sham-operation group (S group), model group (O group), low-dose Yinchenhao decoction group (LY group), and high-dose Yinchenhao decoction group (HY group), with 8 rats in each group. For the rats in the S group, the upper common bile duct was isolated without ligation, and for those in the other groups, double ligation of the middle and upper 1/3 of the common bile duct was performed to establish a model of obstructive jaundice. After 7 days, the rats in the LY group and the HY group were given Yinchenhao decoction by gavage at a dose of 6.3 and 18.9 mL/kg, respectively, while those in the S and O groups were given an equal volume of distilled water by gavage every day for 7 consecutive days, and the rats were treated on day 14. ELISA was used to measure the serum levels of total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), blood urea nitrogen (BUN), and creatinine (Cr); spectrophotometry was used to measure the activity of the oxidative stress factors superoxide dismutase (SOD) and malondialdehyde (MDA) in renal tissue; quantitative real- time PCR and Western blotting were used to measure the mRNA and protein expression levels of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), and NAD(P)H quinone dehydrogenase 1 (NQO1) in renal tissue; immunohistochemistry was used to measure observe the nuclear translocation of Nrf2 protein in renal tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further pairwise comparison within groups. Results Compared with the S group, the O group had significant increases in the levels of TBil, DBil, ALT、GGT, BUN, and Cr, a significant reduction in the activity of SOD, and a significant increase in the level of MDA (all P < 0.05). Compared with the O group, the LY group and the HY group had significant reductions in liver and renal function parameters, a significant increase in the activity of SOD, and a significant reduction in the level of MDA (all P < 0.05). Compared with the S group, the O group had significant reductions in the mRNA and protein expression levels of Nrf2 and NQO1 in renal tissue (all P < 0.05), and compared with the O group, the LY group and the HY group had significant increases in the mRNA and protein expression levels of Nrf2 and NQO1 (all P < 0.05), while there was no significant difference in the protein expression level of Keap1 between groups ( P > 0.05). Compared with the S group, the O group had a significant reduction in the positive rate of Nrf2 in cell nucleus in renal tissue ( P < 0.05), and compared with the O group, the LY group and the HY group had a significant increase in the positive rate of Nrf2 in cell nucleus ( P < 0.05). Conclusion Yinchenhao decoction can effectively alleviate renal injury caused by obstructive jaundice, possibly by upregulating the protein expression of Nrf2 in renal tissue and regulating the nuclear translocation of Nrf2 protein, so as to mediate the protein expression of downstream NQO1, regulate oxidative stress response caused by obstructive jaundice, and thereby alleviate renal injury in rats.

ObjectiveTo determine whether HBV DNA polymerase is associated with T-cell failure and thus mediates the immune escape of HBV-related hepatocellular carcinoma （HCC） tumor cells， and to investigate the specific molecular mechanisms. MethodsLiver cancer cell lines Huh7 and HepG2 stably transfected with HBV DNA polymerase expression plasmid with Flag （Flag-HBV-P） and intercellular adhesion molecule-1 （ICAM1） were co-cultured with Jurkat cells， and MTT assay， qRT-PCR， and ELISA were used to measure Jurkat cell proliferation， activation （CD69 expression）， and secretion of the cytokine IFN-γ. RNA-seq was used to screen for differentially expressed immune-associated molecules between stably transfected cell lines and control cells， and mRNA half-life and protein half-life assays were used to determine the specific levels of the immune-associated molecules that were affected by HBV DNA polymerase. Related websites were used to predict the transcription factors that may bind to the promoter region of this immune-associated molecule， Western blot was used to verify the effect of transcription factors on the immune-associated molecule， and rescue experiment was used to determine whether HBV DNA polymerase affects the expression level of the immune-associated molecule through this transcription factor. The independent-samples t test was used for comparison between two groups. ResultsThe experimental group had significant reductions in Jurkat cell proliferation， activation， and cytokine secretion compared with the control group （all P<0.01）. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had significant reductions in the mRNA and protein expression levels of ICAM1 （all P<0.01）. Website prediction identified the ICAM1 promoter and preliminarily highlighted NFKB1， RELA， and STAT3. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had a significant reduction in the protein expression level of p65 （all P<0.01）. After p65 overexpression， there was a significant increase in the protein expression level of ICAM1， and after the expression of p65 was reduced， there was a significant reduction in the protein expression level of ICAM1 （all P<0.01）. In the rescue experiment， there was no significant difference in the protein expression level of ICAM1 between the control group and the experimental group after p65 overexpression （all P>0.05）. After the overexpression of ICAM1， there were no significant differences in the proliferation， activation， and cytokine secretion of Jurkat cells between the control group and the experimental group （Huh7 and HepG2 cell lines） （all P>0.05）. ConclusionHBV DNA polymerase downregulates the level of ICAM1 to mediate HCC immune escape by inhibiting the expression of p65 in NF-κB.

BACKGROUND: Intervertebral disk (IVD) degeneration, which can cause lower back pain, is a major predisposing factor for disability and can be managed through multiple approaches. However, there is no satisfactory strategy currently available to reconstruct and recover the natural properties of IVDs after degeneration. As tissue engineering develops, scaffolds with embedded cell cultures have proved critical for the successful regeneration of IVDs. METHODS: In this study, an integrated scaffold for IVD replacement was developed. Through scanning electron microscopy and other mechanical measurements, we characterized the physical properties of different hydrogels. In addition, we simulated the physiological structure of natural IVDs. Nucleus pulposus (NP) cells and annulus fibrosusderived stem cells (AFSCs) were seeded in gelatin methacrylate (GelMA) hydrogel at different concentrations to evaluate cell viability and matrix expression. RESULTS: It was found that different concentrations of GelMA hydrogel can provide a suitable environment for cell survival. However, hydrogels with different mechanical properties influence cell adhesion and extracellular matrix component type I collagen, type II collagen, and aggrecan expression. CONCLUSION This tissue-engineered IVD implant had a similar structure and function as the native IVD, with the inner area mimicking the NP tissue and the outer area mimicking the stratified annulus fibrosus tissue. The new integrated scaffold demonstrated a good simulation of disc structure. The preparation of efficient and regeneration-promoting tissueengineered scaffolds is an important issue that needs to be explored in the future. It is hoped that this work will provide new ideas and methods for the further construction of functional tissue replacement discs.

Objective: To investigate the characteristics and prevalence of HIV infection among AIDS volunteer counseling and testing ( VCT ) clients in Shangcheng District, Hangzhou City from 2016 to 2019, so as to provide insights into local AIDS control. Methods: The demographic features, reasons for counseling and detection of anti-HIV antibody were captured from the VCT clinic of Shangcheng Center for Disease Control and Prevention from 2016 to 2019, and the reasons for counseling and prevalence of HIV infections were analyzed. Results: A total of 2 205 clients were included, among whom 1 920 participants ( 87.07% ) were male, 1 094 ( 49.61% ) were at ages of < 30 years, 1 293 ( 58.64% ) were single, and 1 165 ( 52.83% ) had an education level of diploma and above. The common reasons for counseling included a history of homosexual behaviors with men (887 clients, 40.23%), non-commercial irregular heterosexual behaviors ( 661 clients, 29.98% ), commercial heterosexual behaviors ( 308 clients, 13.97% ), HIV-positive spouse/regular sex partners ( 123 clients, 5.58% ), and no high-risk behaviors ( 47 clients, 2.13% ). The positive rate of anti-HIV antibody was 2.95% among the 2 205 clients, and a high positive rate was observed among clients at ages of 50 years and below ( 6.15% ), single clients ( 3.71% ), clients with an education level of primary school and below ( 6.04% ) and clients with HIV-positive spouse/regular sex partners ( 11.38% ). Conclusions Single men at ages of less than 30 years and with a high education level are predominant among VCT clients in Shangcheng District, and a history of homosexual behaviors with men is the predominant reason for counseling. There is a relative low prevalence rate of HIV infection among VCT clients in Shangcheng District; however, a high prevalence rate is found among single visitors at ages of 50 years and older, with an education level of primary school and below and HIV-positive spouse/regular sex partners.

Diabetic nephropathy (DN) is considered the primary causes of end-stage renal disease (ESRD) and is related to abnormal glycolipid metabolism, hemodynamic abnormalities, oxidative stress and chronic inflammation. Antagonism of vascular endothelial growth factor B (VEGF-B) could efficiently ameliorate DN by reducing renal lipotoxicity. However, this pharmacological strategy is far from satisfactory, as it ignores numerous pathogenic factors, including anomalous reactive oxygen species (ROS) generation and inflammatory responses. We found that the upregulation of VEGF-B and downregulation of interleukin-22 (IL-22) among DN patients were significantly associated with the progression of DN. Thus, we hypothesized that a combination of a VEGF-B antibody and IL-22 could protect against DN not only by regulating glycolipid metabolism but also by reducing the accumulation of inflammation and ROS. To meet these challenges, a novel anti-VEGFB/IL22 fusion protein was developed, and its therapeutic effects on DN were further studied. We found that the anti-VEGFB/IL22 fusion protein reduced renal lipid accumulation by inhibiting the expression of fatty acid transport proteins and ameliorated inflammatory responses

Objective: To analyze the characteristics of young students inquiry to voluntary counseling and testing (VCT) clinics, and to provide reference for AIDS prevention and control effectiveness of local schools. Methods: A total of 1 223 students aged 7-28 years were collected and characterized through HIV Testing Consulting Information Management System in Shangcheng VCT clinics during 2016-2020. The epidemic characteristics were analyzed by cross sectional studies. Results: Young students accounted for 25.21%(1 223/4 852) of the total counseling population in VCT clinics from 2016 to 2020. The main reason for counseling was non high risk (28.29%, 346 cases), followed by MSM (26.41%, 323 cases) and non commercial non fixed heterosexual behavior(25.92%, 317 cases). The counseling proportion of non commercial non fixed heterosexual behavior and commercial heterosexual sex behavior declined with time( χ 2 trend =-6.04, -3.59, P <0.01). The average of HIV positive detection rate among young students was 3.03%(37 cases), which increased with time( χ 2 trend=2.45, P <0.01). Conclusion Sex education and HIV/AIDS knowledge education are necessary in primary and secondary schools. Targeted strategies are in great need in terms of MSM behavior intervention.

A randomized, double-blind, active parallel controlled trial will be conducted to include patients with acute upper respiratory infection and wind-warmth lung heat disease (heat in lung-wei pattern). Patients with serious bacterial infection (white blood cell count>1.2×1010, neutrophil>80%) will be excluded.so as to further identify Jinye Baidu Granule indications of Wind-warmth lung heat disease (heat in lung-wei pattern).According to the subjects of symptoms of fever and sore throats,Patients will be divided into three blocks： fever, swelling and sore throat, combined fever and swelling and sore throat. Subjects in three blocks will be divided into treatment group and control group with stratified blocked randomization. The treatment group will be treated with Jinye Baidu Granule, and the control group will be treated with Fufang Shuanghua granule. Primary outcome measure of patients with fever will be body temperature recovery time. Primary outcome measure of patients with swelling and sore throat will be throat symptom score. Primary outcome measures of patients combined with fever and swelling and sore throat will be body temperature recovery time and throat symptom score. Before and after all of the patients in the group were collected blood and urine routine, liver and renal function, electrocardiogram (ECG) data as the safety index while observing it adverse events. A total of 623 patients were included. Compared with control group,sore throat symptoms of patients with sore throat and pharyngeal symptoms total score have significantly different. so Jinny Baidu granule have a significant advantage in the treatment of sore throat. All of the experiments showed that the safety of the drug was good. Nausea, vomiting, stomach pain, diarrhea, digestive tract symptoms may occur only occasionally. We should pay attention to the changes when using in patients with spleen deficiency cold to avoid adverse reaction.

Objective To explore the preventive effects of different extracts from Aspidopterys obcordata on renal tubu-lar epithelial cells injury induced by sodium oxalate in vitro, and initially identify the effective part for treating urolithiasis. Methods The injury model of HK-2 cells induced by sodium oxalate was established to screen the active parts of Aspidopterys obcordata by testing the protective effects of different polarity extracts on HK-2 injury cells through MTT method. Results Dif-ferent extracts from Aspidopterys obcordata improved the activity of HK-2 injury cells, which were elevated to 86.17% and 95.42%by 0. 5 mg?mL-1 and 1 mg?mL-1 aqueous extract, respectively. And the activity reached to 93. 59% and 84. 77% by 0.5 mg?mL-1 and 1 mg?mL-1 50% alcohol extracts, reached above 81.56% by 95% alcohol extracts,all of which showed sig-nificant difference compared with the model group. The HK-2 cells viability were elevated to 82. 53% and 91. 58% by 0.5 mg?mL-1 and 1 mg?mL-1 95% alcohol extracted ethyl acetate parts,and increased to 77.24% and 87.22% by 0.5 mg?mL-1and 1 mg?mL-1 of 95% alcohol isolated n-butanol extracts, approached to 95.46% and 81.36% by 0.5 mg?mL-1 and 1 mg?mL-1 water extracts, all of which showed significant difference compared with the model control group. Conclusion The aqueous extracts and alcohol extracts from Aspidopterys obcordata have obvious preventive effects on HK-2 cells injury, among which the ethyl acetate extracts, n-butanol extracts and water extracts present the remarkable effects, which are supposed to be the active parts for inhibiting calcium oxalate stone formation in vitro.

Objective To study the film￣forming property of film￣forming materials in dragon blood ethanol solution, and provide the reference for screening of film￣forming materials for dragon blood plastics. Methods PVA124,PVA1788, PVP￣K30, EC, MC, PVB were chosen to study the film￣forming property of film￣forming materials based on solubility, formation time, and adhesion. Results PVA1788 and PVP￣K30 mixed in dragon blood ethanol solution in proportion of 4:1 showed good film￣forming property, and the membrane was smooth, uniform and easy to tear off.However, PVA124 and PVP￣K30 mixed in proportion of 4:1 precipitated in dragon blood ethanol solution.EC, MC and PVB were not easy to tear off the membrane. Conclusion PVA1788 and PVP￣K30 mixed in proportion of 4:1 and added with propylene glycol is the optimal choice of film￣forming material for dragon blood spray film.