1.Objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease
Zhaoxi DONG ; Yang SHI ; Jiaming SU ; Yaxuan WEN ; Zheyu XU ; Xinhui YU ; Jie MEI ; Fengyi CAI ; Xinyue ZANG ; Yan GUO ; Chengdong PENG ; Hongfang LIU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(3):398-411
Objective:
To investigate the objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease (DKD).
Methods:
A cross-sectional study enrolled 134 patients with DKD G3-5 stages who met the diagnostic criteria for damp-heat syndrome in DKD. The patients were treated at Dongzhimen Hospital, Beijing University of Chinese Medicine, from May 2023 to January 2024. The patients were divided into three groups: DKD G3, DKD G4, and DKD G5 stage, with 53, 33, and 48 patients in each group, respectively. Clinical general data (gender, age, and body mass index) and damp-heat syndrome scores were collected from the patients. The YZAI-02 traditional Chinese medicine (TCM) AI Tongue Image Acquisition Device was used to capture tongue images from these patients. The accompanying AI Open Platform for TCM Tongue Diagnosis of the device was used to analyze and extract tongue manifestation features, including objective data on tongue color, tongue quality, coating color, and coating texture. Clinical data and objective tongue manifestation characteristics were compared among patients with DKD G3-5 based on their DKD damp-heat syndrome status.
Results:
No statistically significant difference in gender or body mass index was observed among the three patient groups. The DKD G3 stage group had the highest age (P<0.05). The DKD G3 stage group had a lower score for symptoms of poor appetite and anorexia(P<0.05) than the DKD G5 group. No statistically significant difference was observed in damp-heat syndrome scores among the three groups. Compared with the DKD G5 stage group, the DKD G3 stage group showed a decreased proportion of pale color at the tip and edges of the tongue (P<0.05). The DKD G4 stage group exhibited an increased proportion of crimson at the root of the tongue, a decreased proportion of thick white tongue coating at the root, a decreased proportion of pale color at the tip and edges of the tongue, an increased hue value (indicating color tone) of the tongue color in the middle, an increased brightness value (indicating color lightness) of the tongue coating color in the middle, and an increased thickness of the tongue coating (P<0.05). No statistically significant difference was observed in other tongue color proportions, color chroma values, body characteristics, coating color proportions, coating color chroma values, and coating texture characteristics among the three groups.
Conclusion
Tongue features differ in different stages of DKD damp-heat syndrome in multiple dimensions, enabling the inference that during the DKD G5 stage, the degree of qi and blood deficiency in the kidneys, heart, lungs, liver, gallbladder, spleen, and stomach is prominent. Dampness is more likely to accumulate in the lower jiao, particularly in the kidneys, whereas heat evil in the spleen and stomach is the most severe. These insights provide novel ideas for the clinical treatment of DKD.
2.Production of GTKO pigs and kidney xenotransplantation from pigs to rhesus macaques
Yan WANG ; Yue CHANG ; Chang YANG ; Taiyun WEI ; Xiaoying HUO ; Bowei CHEN ; Jiaoxiang WANG ; Heng ZHAO ; Jianxiong GUO ; Hongfang ZHAO ; Xiong ZHANG ; Feiyan ZHU ; Wenmin CHENG ; Hongye ZHAO ; Kaixiang XU ; Ameen Jamal MUHAMMAD ; Zhendi WANG ; Hongjiang WEI
Organ Transplantation 2025;16(4):526-537
Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68+ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.
3.Assessment of the implementation of Radiation shielding requirements for radiotherapy room—Part 4: Radiotherapy room of 252Cf neutron afterloading (GBZ/T 201.4-2015)
Yuze YANG ; Hongfang WANG ; Haoxian YANG ; Quan WU ; Mingsheng LI ; Bala HARI ; Yongzhong MA ; Zechen FENG ; Bin BAI ; Jie GAO ; Wei ZHOU ; Weixu HUANG ; Zhengjie SHI ; Hezheng ZHAI
Chinese Journal of Radiological Health 2025;34(5):660-665
Objective To track and evaluate the implementation and application of the occupational health standard Radiation shielding requirements for radiotherapy room—Part 4: Radiotherapy room of 252Cf neutron afterloading (GBZ/T 201.4-2015) by radiation health technical service agencies, medical institutions, health supervision agencies, and radiotherapy facility design units, and to provide a scientific basis for the further revision and implementation of this standard. Methods Following the Guideline for health standards tracking evaluation (WS/T 536-2017) and the project implementation plan, relevant practitioners were randomly selected for a questionnaire survey. The survey primarily focused on their awareness, standard training, application, and revision suggestions of GBZ/T 201.4-2015. The results were summarized and analyzed. Results A total of 168 evaluation questionnaires were collected from relevant practitioners in 28 provinces. Only 31.6% of the respondents reported being “well familiar” or “ familiar” with the standard, 27.4% of the respondents believed that the standard was widely used, and 45.2% of the respondents believed that the standard could meet the needs of their work. Only 14.9% of the respondents had received relevant training on the standard, more than half of the respondents had not applied the standard within the past 10 years, and 45.2% of the respondents believed that the standard "needs to be revised". Conclusion Due to the small number of californium-252 neutron afterloading radiotherapy devices in operation on the market, the overall awareness of the standard is low, suggesting that relevant authorities need to strengthen training and publicity of the standard, and that certain sections of the standard need to be revised or merged.
4.Aerobic exercise improves blood lipid and blood glucose of obese,diabetic and atherosclerotic rats by re-ducing chemerin
Xiaojing LIN ; Shaoting FU ; Hongfang YANG
Chinese Journal of Rehabilitation Medicine 2024;39(11):1561-1571
0bjective:To confirm the role of decreased chemerin on aerobic exercise-induced improvement of glucose and lipid metabolism using exogenous chemerin in obese,diabetic and atherosclerotic rats,and to explore whether the mechanisms is related to the key enzymes or proteins of glucose and lipid metabolism mediated by peroxi-some proliferator-activated receptor γ(PPARγ)in the peripheral metabolic organs(liver,muscle and fat),includ-ing lipoprotein triglyceride lipase(ATGL)and lipoprotein lipase(LPL)and glucose transporter 4(GLUT4). Method:①Animal experiments:Total of 130 male rats aged 6 weeks were included in this study.In addition to the normal control(Con,n=17)rats,the model rats with obesity(OB),atherosclerosis(AS)and diabetes mellitus(DM)were established by 8-week high-fat diet,8-week high-fat diet combined with intraperitoneal injection of Vitamin D3 or of low dose of streptozotocin,respectively.All the successfully established model rats were ran-domly divided into:the control[AS(n=8),OB(n=8)and DM(n=10)and the exercise(EAS(n=8),EOB(n=8)and EDM(n=10)]rats.All exercise groups were participated in the 4-week moderate-intensity treadmill running exercise with gradually increased load,six days per week and one time per day.Serum chemerin were detect-ed by ELISA.Protein levels of chemerin,CMKLR1,PPARγ,ATGL,LPL and GLUT4 in tissues(liver,gastrocne-mius and perirenal fat)were detected by Western blot.In order to confirm the mediation role of PPARγ,diabet-ic rats were randomly divided into 4 groups:DM(n=10),EDM(n=10),EDM plus PPARγ agonist pioglitazone(EDP,n=10)and EDM plus PPARγ antagonist GW9662(EDG,n=10),and the protein levels of ATGL,LPL and GLUT4 were detected.②Cell experiments:3T3-L1 cells were treated with exogenous chemerin(200 ng/ml)for 48 h,then the cells were collected to detect the protein levels of PPARγ,ATGL,LPL and GLUT4. Result:①The serum chemerin and the protein levels of chemerin and CMKLR1 in tissues(liver,gastrocnemius and perirenal fat)of EAS,EOB and EDM groups rats were significantly decreased after 4-week aerobic exer-cise,with the improvement of blood glucose and lipid.The protein levels of PPARγ,ATGL,LPL in liver,gas-trocnemius and perirenal fat,and GLUT4 in gastrocnemius were significantly increased in the EAS,EOB and EDM rats after 4-week aerobic exercise.② In DM rats,the increase of ATGL,LPL and GLUT4 induced by aerobic exercise were partially reversed by PPARγ inhibitor GW9662 and further enhanced by PPARγ agonist pioglitazone.(3)The protein levels of PPARγ,ATGL,LPL and GLUT4 in 3T3-L1 cells were significantly re-duced after treating the cells with exogenous chemerin. Conclusion:The improved effect of 4 weeks of aerobic exercise on blood glucose and blood lipid in rats with obesity,diabetic and atherosclerosis is related to the reduction of chemerin,thereby upregulating key en-zymes and proteins of glycolipid metabolism(including ATGL,LPL and GLUT4).The effect of chemerin on the above key enzymes and protein is mediated by PPARγ in diabetes rats.
5.Rational selection between internal and external drainage approaches for microinvasive/minimally invasive glaucoma surgery in intraocular pressure control
Hongfang YANG ; Wenhan LU ; Xinghuai SUN
Chinese Journal of Experimental Ophthalmology 2024;42(6):497-502
Glaucoma stands as the leading cause of irreversible blindness, with primary open-angle glaucoma being the predominant type.Although its pathogenic mechanisms are intricate, lowering intraocular pressure remains the most effective treatment.Mainstream glaucoma surgeries aim to lower intraocular pressure by increasing aqueous humor outflow.Over the past two decades, microinvasive/minimally invasive glaucoma surgery (MIGS) has emerged as a hotspot in this field, focusing on minimizing incisions.Domestic practices have followed closely, but clinical outcomes have not been ideal.Based on a comprehensive understanding of the anatomy and pathophysiology of the aqueous outflow pathway and the design principles of various MIGS procedures, ophthalmologists should recognize the advantages and limitations of each technique.It is important to rationally choose between internal and external drainage MIGS and standardize their implementation.In the future, the use of non-invasive multimodal imaging techniques for preoperative assessment of the structural and functional aspects of the eye is expected to facilitate personalized selection and planning for precise treatment.
6.Qidi Tangshen Prescription (QDTS) Regulate Akt1/HIF-1α/Bcl-xl Signaling Pathway to Improve Podocyte Autophagy in Diabetic Nephropathy
Fei GAO ; Huidi XIE ; Borui YU ; Ying ZHOU ; Yang SHI ; Xianhui ZHANG ; Hongfang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):90-97
ObjectiveTo explore the mechanism of Qidi Tangshen prescription (QDTS) in alleviating podocyte injury and reducing urinary protein in diabetic nephropathy (DN). MethodUsing network pharmacology methods, we collected the chemical components and targets of QDTS, as well as the targets related to DN. Subsequently, we constructed a "drug-ingredient-target-disease" network for QDTS in the treatment of DN to systematically elucidate the mechanism. The db/db mice were assigned into the model, QDTS (3.34 g·kg-1), and losartan capsules (10.29 mg·kg-1) groups, and db/m mice served as the normal group. Each group consisted of 8 mice, and they underwent continuous intervention for 8 weeks. After the last administration, mice were euthanized, and the urinary albumin excretion rate (UAER) and renal pathological changes were measured and observed. The expression levels of protein kinase B1 (Akt1), hypoxia-inducible factor-1 alpha (HIF-1α), phosphorylated B-cell lymphoma-extra-large (p-Bcl-xl), as well as autophagy-related indicators microtubule-associated protein 1 light chain 3 (LC3), ubiquitin-binding protein p62 (p62), and autophagy-related gene 6 homolog (Beclin1), were determined. Furthermore, mouse podocytes were divided into the normal glucose (5.5 mmol·L-1), high glucose (35 mmol·L-1), DMSO (35 mmol·L-1 glucose+200 mg·L-1 DMSO), and QDTS (35 mmol·L-1 glucose+200 mg·L-1 QDTS freeze-dried powder) groups. After 48 h of intervention, the protein levels of Akt1, HIF-1α, p-Bcl-xl, LC3, p62, and Beclin1 in podocytes were measured. ResultQDTS had 34 active components acting on 143 targets in the treatment of DN, and 55 targets were related to autophagy, in which Akt1, HIF-1α, and Bcl-xl were the key targets. Compared with the normal group, mice in the model group exhibited significantly increased UAER, glomerular hypertrophy, deposition of blue collagen fibers, thickening of the glomerular basement membrane, and noticeable fusion of podocyte foot processes in some segments. Furthermore, the modeling up-regulated the protein levels of p-Akt1, HIF-1α, and p62 and down-regulating the protein levels of p-Bcl-xl, LC3, and Beclin1 in the renal tissue (P<0.05). Compared with the model group, QDTS and losartan decreased UAER (P<0.05) and alleviated the pathological damage in the renal tissue. Moreover, QDTS and losartan down-regulated the protein levels of p-Akt1, HIF-1α, and p62 and up-regulated the protein levels of p-Bcl-xl, LC3, and Beclin1 in the renal tissue (P<0.05). In comparison to the normal glucose group, the high glucose group displayed up-regulated protein levels of p-Akt1, HIF-1α, and p62 and down-regulated protein levels of p-Bcl-xl, LC3, and Beclin1 in podocytes (P<0.05). Compared with the high glucose group, QDTS down-regulated the protein levels of p-Akt1, HIF-1α, and p62 and up-regulated the protein levels of p-Bcl-xl, LC3, and Beclin1 in podocytes (P<0.05). ConclusionQDTS alleviates podocyte damage and reduced urinary protein in DN by regulating the Akt1/HIF-1α/Bcl-xl signaling pathway, thereby enhancing podocyte autophagy.
7.Implication of Qidi Tangshen Prescription (QDTS) on Podocyte Pyroptosis in Diabetes Nephropathy by Regulating MAPK14/RELA/Caspase-8 Signaling Pathway
Fei GAO ; Borui YU ; Huidi XIE ; Ying ZHOU ; Yang SHI ; Xianhui ZHANG ; Hongfang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):67-75
ObjectiveTo explore the molecular mechanism of Qidi Tangshen prescription (QDTS) in regulating podocyte pyroptosis in diabetes nephropathy (DN). MethodThrough in vivo experiment, db/db mice were divided into the model group, QDTS group (3.34 g·kg-1), valsartan capsule group (10.29 mg·kg-1), with db/m mice serving as the normal control. Each group consisted of 8 mice, and they underwent continuous intervention for 8 weeks. After the last administration, mice were euthanized, and kidney pathological changes were observed. Additionally, the expression levels of pyroptosis-related indicators, including NOD-like receptor protein 3 (NLRP3), Gasdermin D protein (GSDMD), and interleukin-1β (IL-1β) protein, were examined. Through in vitro experiment, mouse podocytes were divided into the normal glucose group (5.5 mmol·L-1 glucose), high glucose group (35 mmol·L-1 glucose), DMSO group (35 mmol·L-1 glucose+200 mg·L-1 DMSO), and QDTS group (35 mmol·L-1 glucose+200 mg·L-1 QDTS freeze-dried powder). After 48 hours of intervention, the expression levels of NLRP3, GSDMD, and IL-1β proteins were measured in podocytes. A drug-ingredient-target-disease interaction network for QDTS in the treatment of DN was constructed by network pharmacology methods. The key signaling pathways regulating podocyte pyroptosis were analyzed, and validation was conducted through in vivo and in vitro experiments. ResultCompared with normal group, glomerular hyperplasia and glomerular basement membrane thickening were observed in model group, and some segments were accompanied by obvious podocellular process fusion. The protein expressions of NLRP3, GSDMD and IL-1β in mouse kidney were increased, the protein expressions of mitogen-activated protein kinase 14 (MAPK14), V-Rel reticuloendotheliosis virus oncogene homology A (RELA) and Caspase-8 in mouse kidney were increased (P<0.05). Compared with model group, kidney pathological injury of mice in QDTS group was significantly reduced, and the expressions of NLRP3, GSDMD and IL-1β in kidney of mice in QDTS group and valsartan group were decreased (P<0.05). The protein expressions of MAPK14, RELA and Caspase-8 in kidney of mice in QDTS group and valsartan group were decreased (P<0.05). Network pharmacology results showed that there were 16 targets for QDTS to regulate DN cell pyrodeath, among which MAPK14, RELA and Caspase-8 were the key targets. Compared with normal glucose group, the protein expressions of NLRP3, GSDMD and IL-1β in high glucose group were increased (P<0.05), and the protein expressions of MAPK14, RELA and Caspase-8 in mouse podocytes were increased (P<0.05). Compared with high glucose group, the expressions of NLRP3, GSDMD and IL-1β in podocytes of mice in QDTS group were decreased (P<0.05), and the expressions of MAPK14, RELA and Caspase-8 in podocytes of mice in QDTS group were decreased (P<0.05). ConclusionQDTS reduces damage to DN podocytes, which is associated with its regulation of the MAPK14/RELA/Caspase-8 signaling pathway and inhibition of podocyte pyroptosis.
8.Total protein level of plasma donors in various regions of China: a survey of 1 373 cases
Yang GAO ; Rong ZHOU ; Hongfang YANG ; Liyi HAN ; Rong HUANG ; Yan HANG ; Demei DONG
Chinese Journal of Blood Transfusion 2024;37(1):58-62
【Objective】 To analyze the influence of plasma donation on human total protein level and the impact of different blood collection tubes on total protein level detection. 【Methods】 A total of 1 373 plasma donors from 11 apheresis plasma stations in 6 provinces/autonomous regions from March to April, 2021 were selected. Whole blood was collected by ordinary blood collection tube without anticoagulant, heparin anticoagulant tube and sodium citrate anticoagulant tube, and then respectively divided into serum group, heparin anticoagulant group, and sodium citrate anticoagulant group. After separating serum and plasma, the samples were subjected to total protein detection using the biuret method. Kruskal-Wallis test was used to compare the total protein levels among different tubes. The plasma donors were divided into male group (n=597) and female group (n=776), and the total protein levels between different genders were compared by t test. The plasma donors were divided into Sichuan group, Hubei group and Gansu group according to the region, and the Games-Howell test was used for comparison. 【Results】 The median serum total protein level of 1 373 donors was 73.1g/L, which was consistent with the reference range of 65-85 g/L. The median total protein levels of the serum group, heparin anticoagulant group and sodium citrate anticoagulant group were 73.1g/L, 73.3g/L and 63.8g/L, respectively, with statistically significant difference (P<0.05). There was statistical significance in total protein level between sodium citrate anticoagulant group and serum group, sodium citrate anticoagulant group and heparin anticoagulant group(P<0.05), but no statistical significance was noticed between serum group and heparin anticoagulant group (P> 0.05). The serum total protein levels of male group and female group were (72.41±5.40)g/L and (73.67±4.95)g/L, reseectively, and the difference was statistically significant (P<0.05). The serum total protein level in Sichuan group, Hubei group and Gansu group was (73.91±4.29)g/L, (74.17±5.11)g/L and (67.09±3.65)g/L, respectively (P<0.05).The difference between Gansu group and Hubei group, Gansu group and Sichuan group was statistically significant (P<0.05), but no significant difference was noticed between Sichuan group and Hubei group (P>0.05). 【Conclusion】 Plasma donors who meet the donation criteria will not experience abnormal total protein levels due to regular plasma donation. There were differences in total protein levels among different blood collection tubes, different genders and different regions. The total protein level of females was higher than that of males. The total protein level was the highest in Hubei province, followed by Sichuan and Gansu.Heparin anticoagulant group was the highest, followed by serum group and sodium citrate anticoagulant group.
9.Predicting the risk of local tumor progression after radiofrequency ablation of hepatocellular carcinoma
Hongfang WANG ; Guanhua YANG ; Minglei WANG ; Yu BAI ; Yong CHEN
Journal of Practical Radiology 2024;40(11):1875-1879
Objective To investigate the risk factors for identifying local tumor progression(LTP)in patients with hepatocellular carcinoma(HCC)after radiofrequency ablation(RFA)and to establish a predictive model.Methods The clinical data of 122 HCC patients treated by RFA were analyzed retrospectively,and then the patients were divided into positive and negative LTP groups according to that whether LTP occurred within 12 months after RFA.The risk factors of LTP were determined using univariate and multivari-ate analysis,and the predictive model was constructed based on these factors and the internal validation was conducted.Results The results of this study showed that multiple number,diameter>2 cm,rough margin,and adjacent to large blood vessels of the tumor could be independent predictors of LTP,which were further incorporated into constructing the predictive model.Internal validation results showed that the area under the curve(AUC)of receiver operating characteristic(ROC)curve was 0.815[95%confidence interval(CI)0.735-0.895],indicating the model with high differentiation ability.The calibration curve was drawn and the Hosmer-Lemeshow goodness-fit test showed that the model had good stability(P>0.05).The decision curve suggested that the model had good clinical application value.Conclusion The independent risk factors of LTP of HCC after RFA are multiple number,diameter>2 cm,rough margin,and adjacent to large blood vessels of the tumor.When the predictive model is integrated with the above factors,it can poten-tially predict the risk of local tumor and may offer useful guidance for individual treatment and follow-up.
10.High-risk phenotypes of genetic disease in a Neonatal Intensive Care Unit population.
Tiantian XIAO ; Qi NI ; Huiyao CHEN ; Huijun WANG ; Lin YANG ; Bingbing WU ; Yun CAO ; Guoqiang CHENG ; Laishuan WANG ; Liyuan HU ; Hongfang MEI ; Yulan LU ; Mengchun GONG ; Xinran DONG ; Wenhao ZHOU
Chinese Medical Journal 2022;135(5):625-627


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