Objective:To determine the contents of four flavonoids by establishing HPLC specific chromatogram for Alpiniae Katsumadai Semen; To evaluate the differences of Alpiniae Katsumadai Semen from different producing areas.Methods:The specific chromatogram was developed on a column of Thermo Acclaim C18 with acetonitrile-0.1% phosphoric acid solution as the mobile phase by gradient elution at a flow rate of 1.0 ml/min. The detective wavelength was 260 nm, and the column temperature was 30 ℃. Similarity evaluation, PCA analysis, and OPLS-DA analysis were conducted. The contents of Alpinetin, Pinocembrin, Cardamonin, Alnustone in 16 batches of Alpiniae Katsumadai Semen.Results:There were 9 characteristic peaks in the specific chromatogram of Alpiniae Katsumadai Semen. Except the sample of S2 (Hainan producing area), the similarity of Alpiniae Katsumadai Semen in different producing areas was greater than 0.90; PCA analysis divided 16 batches of Alpiniae Katsumadai Semen into 2 categories, and OPLS-DA analysis identified 4 differential biomarkers, with the order of impact being peak 3>peak 5>Alpinetin>Cardamonin. Among them, the quality of Alpiniae Katsumadai Semen from Guangdong producing area was generally stable. Moreover, there were significant differences in the contents of Alpinetin and Cardamonin among the indicator components of Alpiniae Katsumadai Semen from different producing areas.Conclusion:This method can effectively analyze the differences in the quality of Alpiniae Katsumadai Semen from different producing areas, providing reference for the quality evaluation of Alpiniae Katsumadai Semen.

Objective:To elucidate the expression of angiopoietin-like protein 4 (ANGPTL4) in LPS-induced septic cardiomyopathy tissue and cardiomyocyte, and to explore the mechanism of ANGPTL4 in septic cardiomyopathy.Methods:Fifty C57BL/6 mice, aged 8 weeks, were randomly(random number) divided into a treatment group (LPS) and a control group ( n = 25 each). The mice in the treatment group were intraperitoneally injected with LPS (10 mg/kg) to establish a sepsis model. After 24 h, the myocardial tissues of the mice in the sepsis group and the control group, which were caused by LPS, were collected for RNA sequencing to pick out the differentially expressed gene of ANGPTL4.Ventricular myocytes of neonatal mice were taken, and the silencing and overexpression vectors of ANGPTL4 were transfected. After 48 hours of transfection, the cells were collected for subsequent detection. Western blot method was used to detect the expression of apoptotic factors Bax, Bcl-2, and Caspase 3 in mouse ventricular myocytes; CCK8 method was used to detect the activity of ventricular myocytes; using the Annexin V-FITC and PI double staining method, the apoptosis of ventricular myocytes was detected. Results:RNA-seq analysis revealed a statistically significant upregulation of ANGPTL4 expression at both transcriptional and translational levels in the ventricular tissue of septic mice, as compared to the control group ( P<0.05). The results of qRT-PCR and Western blot indicated that the mRNA and protein levels of ANGPTL4 in the ventricular tissues and cardiomyocytes of mice treated with LPS were significantly increased ( P<0.05). After transfection of the silencing and overexpression vectors of ANGPTL4 in cardiomyocytes, it was found that compared with NC, the mRNA and protein expression levels of ANGPTL4 in the si-ANGPTL4 group significantly decreased ( P<0.05), the vitality of ventricular myocytes increased ( P<0.05), the expressions of apoptosis-related factors Bax and Caspase 3 significantly decreased ( P<0.05), and the expression of Bcl-2 significantly increased ( P<0.05), and the number of apoptotic cells significantly decreased ( P<0.05); while the transfection of the overexpression vector of ANGPTL4 showed an opposite trend. Conclusions:In septic myocardial tissue and cardiomyocyte, the expression of ANGPTL4 is elevated, resulting in the inhibition of ventricular myocyte viability and the promotion of cardiomyocyte apoptosis.

In order to promote the development of hair transplantation, particularly the establishment of standards, the Hair Transplantation Expert Group of Plastic and Aesthetic National Medical Quality Control Center invited experts in the field of hair transplantation across China and formed a draft of the Expert Consensus on Standard Terminology for Hair Transplantation based on the collation of relevant literature and monographs. By combining Delphi methodology and consensus meetings, the expert group conducted several rounds of consultations on issues such as follicular unit anatomy, harvesting, dissection, and implantation, ultimately presenting 54 terminologies. This consensus considers the original Chinese and English meanings as well as the professional characteristics of terms used in hair transplantation, highlights the unique features and advantages of hair transplantation, and preserves its essence while promoting innovation, with the hope of providing valuable reference for clinical researchers engaged in hair transplantation both domestically and internationally, and better facilitating the standardization and advancement of the field.

Objective:To observe the effects of NDRG1 on proliferation, migration and lumen formation of retinal vascular endothelial cells (RF/6A cells) in monkeys under high glucose condition. Methods:RF/6A cells were divided into normal group, mannitol group, high glucose group, small interfering RNA (siRNA) negative control group without target gene (siRNA group), 30 nmol/L siRNA down-regulated NDRG1 genome (siNDRG1 group) and 50 nmol/L siNDRG1 group. Normal group cells were cultured conventionally. The mannitol group was added with 25 mmol/L mannitol, and the high-glucose group was added with 25 mmol/L glucose. In the siRNA group, 25 mmol/L glucose was added, and then blank siRNA was added for induction. The 30 and 50 nmol/L siNDRG1 groups were added with 25 mmol/L glucose and induced with 30 and 50 nmol/L siRNDRG1, respectively. All cells were incubated for 24 h for follow-up experiments. Cell proliferation was observed by 4', 6-diaminidine 2-phenylindole staining. Cell counting kit-8 staining was used to detect cell activity. The expression level of NDRG1 mRNA and protein was detected by Western blot and real-time quantitative polymerase chain reaction. Cell migration was observed by cell scratch assay. Cell lumen formation assay was used to detect lumen formation. The two-tailed Student t test was used to compare the two groups. One-way analysis of variance was used to compare groups. Results:There were significant differences in cell proliferation rate ( t=36.659, 57.645) mobility rate ( t=24.745, 33.638) and lumen formation number ( t=41.276, 22.867) between high glucose group and normal group and mannitol group ( P <0.01). Compared with normal group and mannitol group, the relative expression levels of NDRG1 gene mRNA and protein in high glucose group were significantly decreased, with statistical significance ( t=46.145, 21.541, 36.738, 32.976; P<0.001). Compared with the siRNA negative group, the relative expression levels of NDRG1 gene mRNA and protein in 30 nmol/L siNDRG1 group and 50 nmol/L siNDRG1 group were significantly decreased, and the differences were statistically significant ( t=44.275, 40.7577, 57.167, 25.877; P<0.01). Compared with normal group and siRNA group, cell mobility in 30 nmol/LsiNDRG1 group was increased, and the difference was statistically significant ( t=57.562, 49.522; P<0.01). Compared with normal group and siRNA group, the number of cell lumen formation in 30 nmol/LsiNDRG1 group was significantly increased in the same field of vision, and the difference was statistically significant ( t=63.446, 42.742; P<0.01). Conclusion:Down-regulation of NDRG1 gene can improve the activity, migration and lumen formation of RF/6A cells under hyperglycemia.

Objective:To observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells.Methods:The experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. Results:In vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance ( t=25.372, 22.419, 25.372; P<0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance ( F=36.821, 38.692, 29.421; P<0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant ( F=39.649, P<0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance ( F=27.472, 22.315, 31.147, 27.472; P<0.05). Conclusion:Over-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.

Objective:To observe the effect of metformin (Met) on inflammatory bodies and focal death in human retinal microvascular endothelial cells (hRMEC) in diabetes mellitus (DM) microenvironment.Methods:Experimental research was divided into in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 9 healthy C57BL/6J male mice were randomly divided into DM group, normal control group, and DM+Met group, with 3 mice in each group. DM group and DM+Met group mice were induced by streptozotocin to establish DM model, and DM+Met group was given Met 400 mg/ (kg · d) intervention. Eight weeks after modeling, the expression of NLRP3, cleaved-membrane perforating protein D (GSDMD) and cleaved-Caspase-1 in the retina of mice in the normal control group, DM group and DM+Met group were observed by immunohistochemical staining. In vitro cell experiments: hRMEC was divided into conventional culture cell group (N group), advanced glycation end products (AGE) group, and AGE+Met group. Joining the AGE, AGE+Met groups cells were induced by 150 μg/ml of glycation end products, and 2.0 mmol/L Met was added to the AGE+Met group. Pyroptosis was detected by flow cytometry; 2' ,7'-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe was used to detect the expression of reactive oxygen species (ROS) in cells of each group. Real-time fluorescence quantitative polymerase chain reaction and Western blot were used to detect the relative mRNA and protein expression levels of NLRP3, cleaved-GSDMD, cleaved-Caspase-1 in each group of cells. Single factor analysis of variance was used for comparison among the three groups.Results:In vivo animal experiments: compared with the DM group, the expression of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in the retina of normal control group and DM+Met group mice was significantly reduced, with significant difference among the 3 groups ( F=43.478, 36.643, 24.464; P<0.01). In vitro cell experiment and flow cytometry showed that the pyroptosis rate of AGE group was significantly higher than that of N group and AGE+Met group ( F=32.598, P<0.01). The DCFH-DA detection results showed that the intracellular ROS levels in the N group and AGE+Met group were significantly lower than those in the AGE group, with the significant difference ( F=47.267, P<0.01). The mRNA ( F=51.563, 32.192, 44.473; P<0.01) and protein levels ( F=63.372, 54.463, 48.412; P<0.01) of NLRP3, cleaved-GSDMD, and cleaved-Caspase-1 in hRMEC of the AGE+Met group were significantly reduced compared to the N group. Conclusion:Met can down regulate the expression of NLRP3 inflammatory body related factors in hRMEC and inhibit pyroptosis.

Objective:To explore the clinical safety and efficacy of transurethral oral mucosa urethroplasty for urethral meatus and navicular fossa stricture reconstruction.Methods:Retrospective analysis of 9 patients who underwent transurethral repair of urethral meatus and navicular fossa stricture by oral mucosa in our hospital from October 2021 to December 2022. The average age was (58.4±10.4) years old. 5 patients had a history of transurethral endoscopic surgery, 2 had penile lichen sclerosis, and 2 had no obvious causes. Nine patients were diagnosed with urethral meatus and navicular fossa stricture through retrograde urethrography before surgery. The average maximum preoperative urine flow rate was(3.2±0.7)ml/s. Surgical procedure: The incision was firstly made at 6 o'clock using ophthalmic scalpel, the entire layer of urethral scar was opened, and gradually penetrated into the urethral cavity until it reached the normal mucosa of the urethra. A fan-shaped wound was obtained by cutting the scar of 4 to 8 o'clock. The enlarged urethral lumen could smoothly pass through the F24 urethral probe. Measure the stricture length and width, and trim the oral mucosa to the appropriate shape. One arm of the 5-0 absorbable suture passed through the tip of the oral mucosal flap and the normal urethral mucosa outside the apex of the urethral fan-shaped wound, and then passed through the skin on the ventral side of the penis. The other arm of the suture passed through the apex of the fan-shaped wound and passes through the skin on the ventral side of the penis. Tighten the suture to bring the oral mucosa into the urethral cavity and cover the wound surface. If the narrow length was longer, we could suture three stitches to fix the oral mucosa with the V-shaped apex of the fan-shaped wound in a similar way, and the rest could be sutured and fixed with the urethral wound edge in direct vision. The actual measured average length of urethral stricture during the surgery was(1.6±0.5) cm. The appearance of the glans penis, stricture recurrence, maximum urine flow rate, and patient's urination symptoms were recorded after surgery 1 to 3 months. Functional success was defined as the lack of patient reported obstructive voiding symptoms, satisfaction with the appearance of the glans penis, and a slit like external urethral orifice.Results:All 9 patients successfully completed the surgery and the average maximum urine flow rate was(21.5±3.7)ml/s after 3 months of follow up. The overall successful rate was 100%.One patient experienced spraying urination 1 month later after removing the catheter. Examination revealed that protrusion and separation were found at the urethral anastomosis, and symptoms disappeared after urethral dilation. The other patients did not have any obvious complications, satisfactory with the appearance of the penis head and urination.Conclusions:Transurethral oral mucosal repair of urethral meatus and navicular fossa stricture could be a safe, and effective surgical method. It not only solves the problem of urination, but also takes into account the cosmetic effect of penis.

This article reports a case of Fournier gangrene after transrectal biopsy of the prostate. A 69-year-old male patient was admitted to our hospital for dysuria. The preoperative diagnosis was bladder stones and suspicious prostate cancer. During the same time, the patient underwent transurethral holmium laser lithotripsy and transrectal prostate biopsy. Fournier gangrene occurred on the second day after the operation. After active treatment such as upgraded antibiotics and debridement of the scrotum and perineum, the condition was controlled. The perineal wound healed 2 months later without secondary suture. Perioperative transurethral examination should be avoided during transrectal prostate biopsy. To avoid delayed diagnosis and treatment, it is necessary to improve the understanding of postoperative complications of Fournier's gangrene.

ObjectiveTo investigate the value of remote consultation of heart sound acquisition in screening and referral of neonates with congenital heart diseases （CHD） in primary hospitals. MethodsA total of 4 030 neonates with non-critical diseases were selected. They were born in Shanghai Pudong New Area Maternal and Child Health Hospital from November 5， 2019 to March 31， 2021. After birth， routine cardiac auscultation was performed and remote consultation of heart sound collection were performed at the same time in combination with percutaneous oxygen saturation measurement to screen CHD. The children with any positive screening index were advised to verify the diagnosis by cardiac ultrasound examination in Shanghai Children's Medical Center. The diagnostic value of different screening methods was compared. ResultsA total of 110 cases were detected positive by routine screening. Among them， 16 cases were lost to follow-up， and 46 cases were confirmed by cardiac ultrasound examination， with a positive diagnosis rate of 48.94% （46/94）. A total of 51 cases were detected positive by routine screening and remote consultation of heart sound collection simultaneously. Among them， 42 cases were confirmed by cardiac ultrasound examination， with a positive diagnosis rate of 82.35% （42/51）. The difference between the two positive diagnosis rates was statistically significant （P<0.001）. ConclusionRemote consultation of heart sound acquisition on the basis of routine neonatal CHD screening can effectively improve the positive diagnosis rate of CHD screening in primary hospitals， and reduce unnecessary referrals. This method is simple and feasible. It has practical value in primary hospitals that lack professional technicians for the diagnosis and treatment of CHD.