OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Hegu" (LI4) and "Taichong" (LR3) on DNA methylation of the solute carrier family 6 member 4 (SLC6A4) gene promoter region in the hippocampus of depressed rats, and to explore the potential antidepressant mechanism of EA. METHODS: Thirty male Sprague-Dawley rats were randomly divided into a blank group, a model group, a medication group, a 5-Azacytidine (5-AZA) group, and an EA group, 6 rats in each group. Depression models were established in the model group, the medication group, the 5-AZA group, and the EA group using chronic unpredictable mild stress (CUMS) combined with solitary housing. The medication group was treated with intragastric administration of fluoxetine hydrochloride capsules; the 5-AZA group was treated with intraperitoneal injection of 5-AZA; the EA group was treated with EA at bilateral "Hegu" (LI4) and "Taichong" (LR3), with disperse-dense wave, frequency of 2 Hz/100 Hz, and intensity of 1-1.2 mA, 20 min each session. All the treatment was given in three groups once daily for 21 consecutive days. Behavioral changes were evaluated by sucrose preference test, open field test, and novelty-suppressed feeding test. Serum levels of serotonin (5-HT), dopamine (DA), and norepinephrine (NE) were measured by ELISA. The expression of SLC6A4 and 5-HT1AR protein and mRNA in hippocampus was detected by Western blot and real-time quantitative PCR, respectively. DNA methylation status of the SLC6A4 promoter region in hippocampal tissue was analyzed by bisulfite sequencing PCR (BSP). RESULTS: Compared with the blank group, the model group showed decreased sucrose preference, reduced total locomotor distance, and prolonged latency to feeding (P<0.05), decreased serum 5-HT, DA, and NE levels (P<0.05), downregulated hippocampal SLC6A4 and 5-HT1AR protein and mRNA expression (P<0.05), and increased CpG site methylation rate of the SLC6A4 promoter region (P<0.05). Compared with the model group, the medication group, the 5-AZA group, and the EA group exhibited increased sucrose preference, increased total locomotor distance, shortened latency to feeding (P<0.05), elevated serum 5-HT, DA, and NE levels (P<0.05), upregulated hippocampal SLC6A4 and 5-HT1AR protein and mRNA expression (P<0.05), and reduced CpG site methylation rate of the SLC6A4 promoter (P<0.05). Compared with the medication group and the 5-AZA group, the EA group showed higher sucrose preference, greater total locomotor distance, shorter latency to feeding (P<0.05), and increased serum DA and NE levels (P<0.05). CONCLUSION EA could improve depressive behaviors in depressed rat models. The underlying mechanism may involve inhibition of SLC6A4 hypermethylation in the hippocampus on the serotonergic system, upregulation of SLC6A4 and 5-HT1AR protein and mRNA expression, and elevation of monoamine neurotransmitters such as 5-HT.
Animals ; Electroacupuncture ; Male ; Hippocampus/metabolism* ; Rats, Sprague-Dawley ; Rats ; Acupuncture Points ; DNA Methylation ; Depression/metabolism* ; Promoter Regions, Genetic ; Serotonin Plasma Membrane Transport Proteins/metabolism* ; Humans

OBJECTIVES: To investigate the effects of early life intervention with Bifidobacterium bifidum BD-1 (B. bifidum BD-1) on hyperactivity in a female mouse model of attention deficit hyperactivity disorder (ADHD) and explore the underlying mechanisms. METHODS: Eight newborn female Wistar-Kyoto (WKY) rats and 6 spontaneous hypertensive rats (SHRs) were gavaged with saline and another 6 SHRs were gavaged with B. bifidum BD-1 (10⁹ CFU) daily for 3 weeks. Open field test of the rats was conducted at 7 weeks, and fecal samples were collected at weaning (3 weeks) and at 7 weeks for 16S rRNA sequencing. Immunofluorescent staining was used to detect dopamine transporter (DAT) and tyrosine hydroxylase (Th) levels in the striatum and activated microglia in the prefrontal cortex. Treg cells in the mesenteric lymph nodes, spleen and blood were analyzed using flow cytometry. RESULTS: The SHRs traveled a significantly greater distance in open fields test than WKY rats, and this behavior was significantly attenuated by B. bifidum BD-1 intervention. The expression of DAT and Th in the striatum was significantly lower in the SHRs than in WKY rats, while B. bifidum BD-1 treatment obviously increased Th levels in the SHRs. B. bifidum BD-1 intervention significantly deceased the number of activated microglia and increased Treg cell counts in the spleen of SHRs. The treatment also enhanced α diversity in gut microbiota of the SHRs and resulted in a decreased Firmicutes/Bacteroidota ratio, more active Muribaculaceae growth, and suppression of Clostridia_UCG-014 proliferation. CONCLUSIONS Early life intervention with B. bifidum BD-1 alleviates hyperactivity in female SHRs by modulating the gut microbiota and peripheral immune response, suppressing neuroinflammation and improving dopaminergic system function. These findings provide evidence for early prevention strategies and support the development and application of psychobiotics for ADHD.
Animals ; Female ; Rats ; Rats, Inbred WKY ; Rats, Inbred SHR ; Attention Deficit Disorder with Hyperactivity/therapy* ; Bifidobacterium bifidum ; Probiotics/therapeutic use* ; Dopamine Plasma Membrane Transport Proteins/metabolism* ; Tyrosine 3-Monooxygenase/metabolism* ; Gastrointestinal Microbiome ; Disease Models, Animal

OBJECTIVE: To investigate the feasibility and prognostic implications of assessing volume status during early fluid resuscitation in septic patients based on estimated plasma volume status (ePVS). METHODS: A prospective study was conducted. Patients with sepsis admitted to intensive care unit (ICU) of the First Affiliated Hospital of Kunming Medical University from March to December in 2023 were enrolled. The general information and laboratory indicators at ICU admission were recorded, and ePVS, sequential organ failure assessment (SOFA) score, acute physiology and chronic health status evaluation II (APACHE II) score were calculated. The vital signs, arterial blood gas analysis and volume status related indicators before liquid resuscitation (T0h) and 3 hours (T3h) and 6 hours (T6h) of fluid resuscitation were recorded. The diameter and variability of the inferior vena cava (IVC) were measured by ultrasound, and ePVS, percentage change value of estimated plasma volume status (ΔePVS%), difference in central venous-to-arterial partial pressure of carbon dioxide (Pcv-aCO₂), and lactate clearance rate (LCR) were calculated. Patients were divided into sepsis group and septic shock group based on the diagnosis at ICU admission, and septic patients were subdivided into survival group and death group based on their 28-day survival status. The differences in clinical data between the groups were compared. The correlation between ePVS or ΔePVS% and volume status related indicators during early liquid resuscitation was analyzed by Spearman rank sum correlation test. The predictive value of each variable for 28-day survival in patients with sepsis was analyzed by receiver operator characteristic curve (ROC curve), and 28-day death risk factors were analyzed by Logistic regression method. RESULTS: Fifty-four septic patients were enrolled in the final analysis, including 17 with sepsis and 37 with septic shock; 34 survived at 28 days and 20 died, with a 28-day survival rate of 63.0%. Compared with the sepsis group, the septic shock group had a lower venous ePVS at ICU admission [dL/g: 4.96 (3.67, 7.15) vs. 7.55 (4.36, 10.07), P < 0.05]. Compared with the death group, the survival group had higher T6h arterial and venous ΔePVS%, and albumin [Alb; T6h arterial ΔePVS% (%): 11.57% (-1.82%, 31.35%) vs. 0.48% (-5.67%, 6.02%), T6h venous ΔePVS%: 9.62% (3.59%, 25.75%) vs. 1.52% (-9.65%, 7.72%), Alb (g/L): 27.57±4.15 vs. 23.77±6.97, all P < 0.05], lower SOFA score, APACHE II score, AST, T0h Lac, and T3h and T6h norepinephrine dosage [SOFA score: 9.00 (8.00, 10.00) vs. 11.50 (9.25, 14.50), APACHE II score: 18.00 (14.75, 21.25) vs. 25.50 (21.00, 30.00), AST (U/L): 34.09 (23.20, 56.64) vs. 79.24 (25.34, 196.59), T0h Lac (mmol/L): 1.75 (1.40, 2.93) vs. 3.25 (2.33, 5.30), norepinephrine dosage (mg): 0.98 (< 0.01, 3.10) vs. 4.60 (1.05, 8.55) at T3h, 1.82 (0.38, 5.30) vs. 8.20 (2.80, 17.73) at T6h, all P < 0.05]. While there were no significantly differences in other basic data and ePVS at all of the time points before and after resuscitation between the two groups. Correlation analysis showed that T6h venous ePVS was significantly positively correlated with T6h IVC variability in septic patients (r = 0.360, P < 0.05), T0h arterial ePVS was significantly negatively correlated with T3h and T6h liquid intake volume (r₁ = -0.367, r₂ = -0.280, both P < 0.05), and venous ePVS at ICU admission was significantly positively correlated with NT-proBNP at ICU admission (r = 0.409, P < 0.05). T6h venous ΔePVS% was significantly positively correlated with T3h liquid intake volume and T6h LCR (r₁ = 0.286, r₂ = 0.286, both P < 0.05), and significantly negatively correlated with T6h urine volume and T6h change value of Pcv-aCO₂ (ΔPcv-aCO₂; r₁ = -0.321, r₂ = -0.371, both P < 0.05). ROC curve analysis showed that the area under the ROC curve (AUC) of T6h venous ΔePVS% for predicting 28-day survival in septic patients was 0.726 [95% confidence interval (95%CI) was 0.578-0.875, P = 0.006], with a sensitivity of 82.4%, a specificity of 60.0%, and an optimal cut-off value of 3.09%. Binary multifactorial Logistic regression analysis showed that an increase in T6h venous ΔePVS% was a protective factor for 28-day death in patients with sepsis on early fluid resuscitation [odds ratio (OR) = 0.900, 95%CI was 0.834-0.972, P = 0.007]. CONCLUSIONS ePVS may have potential for assessing the volume status of septic patients during early fluid resuscitation. The ΔePVS% during early fluid resuscitation may help to identify septic patients with a poor prognosis.
Humans ; Prognosis ; Fluid Therapy ; Sepsis/physiopathology* ; Prospective Studies ; Plasma Volume ; Intensive Care Units ; Resuscitation ; Male ; Female ; Middle Aged ; Shock, Septic/therapy*

BACKGROUND

Platelet rich plasma (PRP) is increasingly used as an adjunct treatment for androgenetic alopecia (AGA). PRP preparation involves anticoagulant tubes, such as PRP kits or regular tubes such as EDTA. Studies have shown that platelet concentration in PRP correlates with the growth factors present. However, there is limited data regarding the platelet yield across different anticoagulant tubes. This study will aid in the use of EDTA tube as an alternative to the commercially available PRP kits.

This study aims to determine the platelet concentration level obtained in EDTA tubes versus the commercially available PRP kit in patients with AGA.

This was a cross-sectional, quantitative study in 27 adult males with AGA. The venous blood extracted were placed in EDTA and PRP kit tubes and were centrifuged. A hemoanalyzer was used to obtain the platelet concentration levels. Statistical analyses using paired t-test was performed using the STATA MP Software.

Comparative analyses indicated that the mean difference in platelet concentration between the PRP kit and EDTA was -472.67 (95% Cl = -575.40 to - 369.93). Thus, the mean platelet concentration (t=-9.46, p=0.001) was statistically different between the two groups with a significantly higher mean platelet concentration with EDTA as compared to the PRP kit.

The platelet concentration levels obtained from the EDTA tubes were higher than the PRP kit. Thus, the EDTA tube may be a good alternative to the costly commercially available PRP kit.

Objective:To analyze the efficacy, influencing factors, and risk warning of multi-plane low-temperature plasma radiofrequency ablation（MLT-RFA） in the treatment of obstructive sleep apnea hypopnea syndrome（OSAHS）. Methods:A total of 118 OSAHS patients admitted from October 2022 to June 2024 were selected as the research subjects. They were divided into mild group（n=46）, moderate group（n=52）, and severe group（n=20） according to the severity of their condition. MLT-RFA treatment was used for all patients. After surgery, the results of polysomnography（PSG） and the changes in the Calier Sleep Apnea Quality of Life Index（SAQLI） were observed before and after treatment. The incidence of complications after treatment was recorded, and the clinical efficacy of the patients was evaluated. At the same time, they were divided into a treatment effective group（n=106） and an ineffective group（n=12） according to their effects. The general clinical data of the two groups were compared, and binary logistics regression analysis was conducted to identify independent factors that affect treatment efficacy and construct a model. ROC curve analysis was used to evaluate the diagnostic efficacy of the model. Results:The treatment effectiveness rate of the mild group was 93.48%, the moderate group was 90.38%, and the severe group was 80.00%. There was no statistically significant difference in the treatment effectiveness rate among the three groups（P>0.05）. The AHI of the mild group, moderate group, and severe group increased sequentially, while the LSaO2and SAQLI scores decreased sequentially. After treatment, the AHI of all three groups decreased compared to before treatment, while the LSaO2and SAQLI scores increased compared to before treatment, and the differences were statistically significant（P<0.05）. The pre-treatment AHI of the effective group was lower than that of the ineffective group, and the pre-treatment LSaO2and SAQLI were higher than those of the ineffective group, with statistically significant differences（P<0.05）. Pre-treatment LSaO2and pre-treatment SAQLI are independent factors affecting the efficacy of MLT-RFA（P<0.05）. The AUC of pre-treatment LSaO2, pre-treatment SAQLI, and combined prediction were 0.907, 0.763, and 0.947, respectively, with sensitivities of 0.896, 0.840, and 0.917, and specificities of 0.833, 0.667, and 0.887, respectively. Conclusion:MLT-RFA has a significant effect on the treatment of OSAHS, and the AHI, LSaO2, and SAQLI of patients before treatment can predict the treatment effect, with LSaO2 and SAQLI being independent influencing factors. The combinerd prediction model exhibits high diagnostic efficiency, sensitivity, and specificity.
Radiofrequency Ablation/methods* ; Plasma Gases ; Sleep Apnea, Obstructive/surgery* ; Polysomnography ; Postoperative Complications/epidemiology* ; Quality of Life ; Severity of Illness Index ; Treatment Outcome ; Humans

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

The etiology of nervous system diseases is complicated, posing significant harm to patients and often resulting in poor prognoses. In recent years, the role of dopaminergic system in nervous system diseases has attracted much attention, and its complex regulatory mechanism and therapeutic potential have been gradually revealed. This paper reviews the role of dopaminergic neurons, the neurotransmitter dopamine, dopamine receptors and dopamine transporters in neurological diseases (including Alzheimer's disease, Parkinson's disease and schizophrenia), with a view to further elucidating the disease mechanism and providing new insights and strategies for the treatment of neurological diseases.
Humans ; Dopamine/metabolism* ; Nervous System Diseases/physiopathology* ; Parkinson Disease/physiopathology* ; Receptors, Dopamine/metabolism* ; Dopaminergic Neurons/physiology* ; Dopamine Plasma Membrane Transport Proteins/metabolism* ; Alzheimer Disease/physiopathology* ; Schizophrenia/physiopathology* ; Animals

In the clinical stage, suspected hemolytic plasma may cause hemolysis illness, manifesting as symptoms such as heart failure, severe anemia, etc. Applying a deep learning method to plasma images significantly improves recognition accuracy, so that this paper proposes a plasma quality detection model based on improved "You Only Look Once" 5th version (YOLOv5). Then the model presented in this paper and the evaluation system ‌were introduced‌ into the plasma datasets, and ‌the average accuracy of the final classification reached 98.7%‌. The results of this paper's experiment were obtained through the combination of several key algorithm modules including‌ omni-dimensional dynamic convolution, pooling with separable kernel attention, residual bi-fusion feature pyramid network, ‌and‌ re-parameterization convolution. The method of this paper‌ obtains the feature information of spatial mapping efficiently, and enhances the average recognition accuracy of plasma quality detection. This paper presents a high-efficiency detection method for plasma images, aiming to provide a practical approach to prevent hemolysis illnesses caused by external factors.
Algorithms ; Humans ; Hemolysis ; Plasma ; Deep Learning ; Image Processing, Computer-Assisted/methods*

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 effectiveness of arthroscopic superior capsular reconstruction using a "sandwich" patch combined with platelet-rich plasma (PRP) injection in treating massive irreparable rotator cuff tears. METHODS: A clinical data of 15 patients (15 sides) with massive irreparable rotator cuff tears, who were admitted between September 2020 and March 2023 and met the selective criteria, was retrospectively analyzed. There were 8 males and 7 females with an average age of 62.1 years (range, 40-80 years). The rotator cuff tears were caused by trauma in 7 cases and other reasons in 8 cases. The disease duration ranged from 5 to 25 months, with an average of 17.7 months. According to the Hamada grading, the rotator cuff tears were rated as grade 1 in 2 cases, grade 2 in 8 cases, and grade 3 in 5 cases. All patients were underwent superior capsular reconstruction using the "sandwich" patches (autologous fascia lata+polypropylene patch+autologous fascia lata) combined with PRP injection on patches. The pre- and post-operative active range of motion (ROM) of the shoulder joint, American Shoulder and Elbow Surgeons (ASES) score, Constant-Murley score, University of California, Los Angeles Shoulder Rating Scale (UCLA) score, and visual analogue scale (VAS) score were recorded. The subacromial space was measured on the imaging and rotator cuff integrity was assessed based on Sugaya grading. RESULTS: All incisions healed by first intention after operation without any complications such as infection. All patients were followed up 12-18 months (mean, 14.4 months). At last follow-up, the active ROMs of flexion, abduction, external rotation, internal rotation of the shoulder joint, subacromial space, ASES score, Constant-Murley score, and UCLA score increased, and VAS score decreased, showing significant differences when compared with preoperative values ( P<0.05). There was no significant difference in the Sugaya grading between last follow-up and immediately after operation ( P>0.05). CONCLUSION For massive irreparable rotator cuff tears, arthroscopic superior capsular reconstruction using the "sandwich" patches combined with PRP injection can restore stability of the shoulder joint, relieve pain, promote rotator cuff healing, and achieve good short-term effectiveness.
Humans ; Platelet-Rich Plasma ; Female ; Male ; Middle Aged ; Aged ; Rotator Cuff Injuries/therapy* ; Arthroscopy/methods* ; Adult ; Retrospective Studies ; Aged, 80 and over ; Treatment Outcome ; Plastic Surgery Procedures/methods* ; Rotator Cuff/surgery* ; Range of Motion, Articular ; Shoulder Joint/surgery*