ObjectiveThis paper aims to study the potential active compound components and action mechanism of Shenqi Dihuangtang in the treatment of diabetic kidney disease （DKD） through network pharmacology and in vivo experimental verification. MethodsUltra-high-performance liquid chromatography-Q-exactive orbitrap mass spectrometry （UPLC-Q-Exactive Orbitrap MS） technology was used to clarify the main active chemical components of Shenqi Dihuangtang， and it was combined with network pharmacology methods such as gene ontology （GO） functional annotations and Kyoto encyclopedia of genes and genome （KEGG） to predict the potential action mechanism of Shenqi Dihuangtang in treating DKD. Subsequently， the DKD model of db/db male mice was established， and the mice were randomly divided into model group， low-dose Shenqi Dihuangtang group （6.10 g·kg^-1）， medium-dose Shenqi Dihuangtang group （12.19 g·kg^-1）， high-dose Shenqi Dihuangtang group （24.38 g·kg^-1）， and daplizin group （1.25 mg·kg^-1）. During the same period， C57BL/6J male mice were selected into normal group and received drug intervention for 8 weeks， respectively. During this period， the body weight and fasting blood glucose （FBG） of the mice were dynamically monitored， and oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were performed at the end of dosing. The levels of serum creatinine （SCr）， blood urea nitrogen （BUN）， uric acid （UA）， albumin （ALB）， and total protein （TP） were measured by an automatic biochemical analyzer， and 24-hour urine protein was measured by a urine protein quantitative kit. Hematoxylin-eosin （HE）， periodic-acid Schiff （PAS）， and Masson staining were employed to observe the renal histopathology. The expression of nephrotic protein Nephrin was observed by immunohistochemistry. Western blot was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins such as TGF-β₁， Smad2/3， α-smooth muscle actin （α-SMA）， neural-cadherin （N-Cadherin）， and snail protein. ResultsUPLC-Q-Exactive Orbitrap MS identified 384 active compounds in the aqueous extract of Shenqi Dihuangtang. According to oral bioavailability≥30% and the five drug-like principles， 44 key active ingredients were screened out， and 169 intersection targets highly correlated with DKD were matched. Among them， there was a significant interaction relationship between tumor necrosis factor（TNF）， interleukin（IL）-6， protein kinase B（Akt）1， Caspase-3， Jun proto-oncogene （JUN）， hypoxia inducible factor-1α（HIF-1α）， B cell lymphoma-2（Bcl-2）， matrix metallopeptidase-9（MMP-9）， IL-1β， and TGF-β₁. GO functional annotations were significantly enriched in cellular components such as membrane rafts， membrane microdomains， and collagen-containing extracellular matrix， molecular functions such as DNA-binding transcription factor binding， R-Smad binding， and Smad protein binding， as well as biological processes such as reactions to lipopolysaccharides（LPS）， reactions to bacteria-derived molecules， and wound healing. The KEGG pathway was significantly enriched in lipids and atherosclerosis， TGF-β signaling pathway， phosphatidylinositol 3 kinase （PI3K）/Akt signaling pathway， etc. In vivo experimental results showed that the high-dose Shenqi Dihuangtang group could significantly reduce FBG levels in db/db mice （P<0.01）， improve OGTT （P<0.01） and ITT （P<0.01） levels， reduce SCr （P<0.01）， BUN （P<0.01）， UA （P<0.01） and 24-hour BUN （P<0.01）， and increase ALB （P<0.01） and TP （P<0.01） levels. Pathological staining confirmed that the high-dose Shenqi Dihuangtang group could significantly reduce the glomerular mesangial matrix area and collagen deposition （P<0.01） and upregulate the positive expression rate of Nephrin （P<0.01）. Western blot results showed that the high-dose Shenqi Dihuangtang group significantly downregulated the expression of TGF-β₁ （P<0.01） and Smad2/3 （P<0.01） signal molecules and inhibited the protein levels of α-SMA （P<0.01）， N-Cadherin （P<0.01）， and Snail （P<0.01）. ConclusionShenqi Dihuangtang can inhibit the TGF-β₁/Smad signaling axis and block the renal EMT process， thereby improving DKD renal fibrosis damage. Further analysis of its key active components and clinical transformation pathways is needed in the future.

Objective To compare the early clinical efficacy of arthroscopic bridge suture combined with strengthening thread fixation for posterior cruciate ligament(PCL)tibial insertion avulsion frac-tures and traditional open double-row anchor suture fixation,and to explore a safe,effective,and min-imally invasive reduction and fixation method for PCL tibial insertion fractures in knee joint under ar-throscopy.Methods This study was a retrospective study.Sixty patients with PCL tibial insertion frac-tures(Meyers&McKeever types II-IV)and admitted to the Department of Orthopedics of Kunming Hospital of Traditional Chinese Medicine between June 2021 and June 2023 were included,and ran-domly divided into a traditional group(age:39.56±3.24 years,male/female:16/14)and a minimally in-vasive group(age:40.32±4.38,male/female:18/12),each of 30,according to their admission or-der.All patients were of Grade III positive in the drawer test before surgery.The traditional group un-derwent posterior medial inlay incision,reduction under direct vision and double-row anchor suture fix-ation,while the minimally invasive group underwent reduction under total arthroscopy,suture-bridge suture at the tendon-bone junction of PCL,and one anchor suture placed at the femoral insertion of PCL,with the anchor suture line serving as the reinforcing suture.Then,the anchor and suture-bridge suture lines were respectively pulled into the two tunnels behind the tibia from the front and then pulled out from the front of the tibia,and the reinforcing suture was knotted in front of the tib-ia.The bridge suture line was fixed in front of the tibia with an external row anchor.Six months after surgery,both groups were evaluated the efficacy using the posterior drawer test and Lysholm score.Re-sults At the follow-up six months after the surgery,both groups had good fracture healing,with one wound infection and delayed healing in the traditional group and total one-stage heal in the other group.According to the physical examination 6 months after surgery,all were negative in the posterior drawer test,except one of degree II positive in the traditional group.Moreover,the average Lysholm score of the traditional group increased from 65.23±3.48 before surgery to 87.64±4.58 points after surgery(P<0.05),while that of the minimally invasive group increased from 64.35±2.52 to 86.82±2.58(P<0.05),showing no significant differences between the two groups(P>0.05).Conclusion Both techniques in this study can achieve excellent surgical outcomes in the treatment of posterior cruciate ligament avulsion fractures at the tibial insertion site.

Objective:To investigate the effect of acupoint massage combined with oral immune therapy (C-OIT) on oral feeding and growth and development of very low birth weight infants, so as to provide reference for healthy growth of very low birth weight infants.Methods:A prospective randomized controlled study was conducted. The very low birth weight infants admitted to the neonatal intensive care unit, Affiliated Hospital of Jining Medical University from January 2020 to December 2021 were selected using the convenience sampling method. They were divided into group A and group B using the random number table method, with 150 cases in each group. Group A received routine nursing and C-OIT. On this basis, group B received acupoint massage. Oral feeding, growth and development, and other indexes were compared between the two groups.Results:Finally, 150 cases were included in group A and group B, respectively. In group A, there were 78 male cases and 72 female cases, with gestational age of (30.49 ± 1.57) weeks. In group B, there were 74 male cases and 76 female cases, with gestational age of (30.61 ± 1.63) weeks. The time for abdominal distension to disappear, time to oral feeding initiation, time to full oral feeding and gastric tube indwelling time in group B [(4.35 ± 2.19), (4.26 ± 1.02), (19.03 ± 1.84), (6.27 ± 1.23) d] were shorter than those in group A[(8.03 ± 2.34), (8.63 ± 1.74), (20.49 ± 1.62), (9.34 ± 1.85) d], and the differences were statistically significant ( t values were 7.29-26.54, all P<0.05). The total score of the Preterm Oral Feeding Readiness Scale in group B (32.49 ± 1.52) was higher than that in group A (29.40 ± 3.14), and the difference was statistically significant ( t=10.84, P<0.05). At discharge, the difference values of weight, head circumference and height of group B [(0.92 ± 0.10) kg, (4.31 ± 0.61) cm and (4.08 ± 0.53) cm] were higher than those of group A[(0.81 ± 0.09) kg, (3.47 ± 0.57) cm and (3.81 ± 0.42) cm], and the differences were statistically significant ( t=10.01, 12.32, 4.89, all P<0.05). The length of hospitalization in group B (30.26 ± 4.91) d was shorter than that in group A (38.54 ± 5.27) d, and the difference was statistically significant ( t=14.07, P<0.05). Conclusions:Acupoint massage combined with C-OIT can shorten the time to oral feeding, and improve feeding intolerance of very low birth weight infants. It is expected to improve the growth and development status and quality of life of very low birth weight infants in early stage.

Objective To compare the early clinical efficacy of arthroscopic bridge suture combined with strengthening thread fixation for posterior cruciate ligament(PCL)tibial insertion avulsion frac-tures and traditional open double-row anchor suture fixation,and to explore a safe,effective,and min-imally invasive reduction and fixation method for PCL tibial insertion fractures in knee joint under ar-throscopy.Methods This study was a retrospective study.Sixty patients with PCL tibial insertion frac-tures(Meyers&McKeever types II-IV)and admitted to the Department of Orthopedics of Kunming Hospital of Traditional Chinese Medicine between June 2021 and June 2023 were included,and ran-domly divided into a traditional group(age:39.56±3.24 years,male/female:16/14)and a minimally in-vasive group(age:40.32±4.38,male/female:18/12),each of 30,according to their admission or-der.All patients were of Grade III positive in the drawer test before surgery.The traditional group un-derwent posterior medial inlay incision,reduction under direct vision and double-row anchor suture fix-ation,while the minimally invasive group underwent reduction under total arthroscopy,suture-bridge suture at the tendon-bone junction of PCL,and one anchor suture placed at the femoral insertion of PCL,with the anchor suture line serving as the reinforcing suture.Then,the anchor and suture-bridge suture lines were respectively pulled into the two tunnels behind the tibia from the front and then pulled out from the front of the tibia,and the reinforcing suture was knotted in front of the tib-ia.The bridge suture line was fixed in front of the tibia with an external row anchor.Six months after surgery,both groups were evaluated the efficacy using the posterior drawer test and Lysholm score.Re-sults At the follow-up six months after the surgery,both groups had good fracture healing,with one wound infection and delayed healing in the traditional group and total one-stage heal in the other group.According to the physical examination 6 months after surgery,all were negative in the posterior drawer test,except one of degree II positive in the traditional group.Moreover,the average Lysholm score of the traditional group increased from 65.23±3.48 before surgery to 87.64±4.58 points after surgery(P<0.05),while that of the minimally invasive group increased from 64.35±2.52 to 86.82±2.58(P<0.05),showing no significant differences between the two groups(P>0.05).Conclusion Both techniques in this study can achieve excellent surgical outcomes in the treatment of posterior cruciate ligament avulsion fractures at the tibial insertion site.

Objective:To investigate the effect of acupoint massage combined with oral immune therapy (C-OIT) on oral feeding and growth and development of very low birth weight infants, so as to provide reference for healthy growth of very low birth weight infants.Methods:A prospective randomized controlled study was conducted. The very low birth weight infants admitted to the neonatal intensive care unit, Affiliated Hospital of Jining Medical University from January 2020 to December 2021 were selected using the convenience sampling method. They were divided into group A and group B using the random number table method, with 150 cases in each group. Group A received routine nursing and C-OIT. On this basis, group B received acupoint massage. Oral feeding, growth and development, and other indexes were compared between the two groups.Results:Finally, 150 cases were included in group A and group B, respectively. In group A, there were 78 male cases and 72 female cases, with gestational age of (30.49 ± 1.57) weeks. In group B, there were 74 male cases and 76 female cases, with gestational age of (30.61 ± 1.63) weeks. The time for abdominal distension to disappear, time to oral feeding initiation, time to full oral feeding and gastric tube indwelling time in group B [(4.35 ± 2.19), (4.26 ± 1.02), (19.03 ± 1.84), (6.27 ± 1.23) d] were shorter than those in group A[(8.03 ± 2.34), (8.63 ± 1.74), (20.49 ± 1.62), (9.34 ± 1.85) d], and the differences were statistically significant ( t values were 7.29-26.54, all P<0.05). The total score of the Preterm Oral Feeding Readiness Scale in group B (32.49 ± 1.52) was higher than that in group A (29.40 ± 3.14), and the difference was statistically significant ( t=10.84, P<0.05). At discharge, the difference values of weight, head circumference and height of group B [(0.92 ± 0.10) kg, (4.31 ± 0.61) cm and (4.08 ± 0.53) cm] were higher than those of group A[(0.81 ± 0.09) kg, (3.47 ± 0.57) cm and (3.81 ± 0.42) cm], and the differences were statistically significant ( t=10.01, 12.32, 4.89, all P<0.05). The length of hospitalization in group B (30.26 ± 4.91) d was shorter than that in group A (38.54 ± 5.27) d, and the difference was statistically significant ( t=14.07, P<0.05). Conclusions:Acupoint massage combined with C-OIT can shorten the time to oral feeding, and improve feeding intolerance of very low birth weight infants. It is expected to improve the growth and development status and quality of life of very low birth weight infants in early stage.

This study was performed to screen the sequence of specific nanobody targeting SARS-CoV-2 S protein,and to express and purify recombinant anti-SARS-CoV-2 nanobodies,and evaluate their characteristics and application potential.Based on the previously constructed natural bacteriophage nanobody display library,the anti-SARS-CoV-2 S protein nanobody sequences were screened,with biotinylated SARS-CoV-2 S protein receptor-binding domain antigen as target,by biotin-streptavidin liquid phase screening method,phage-ELISA and sequencing.Recombinant anti-SARS-CoV-2 S protein nanobody expression vectors were constructed,the protein was induced by IPTG and then purified,and their characteristics and application potential were analyzed by SDS-PAGE,Westem blot and ELISA methods.Two anti-SARS-CoV-2 S protein nanobody sequences were successfully obtained,and the corresponding prokaryotic expression bacteria were constructed.The recombinant anti-SARS-CoV-2 S protein nanobody was expressed and purified successfully,which can specifically bind to SARS-CoV-2 S protein.In conclusion,the recombinant anti-SARS-CoV-2 S protein nanobodies have screened and expressed successfully,which provides a basis for its application in SARS-CoV-2 detection and treatment,and facilitates related researches.

Objective:To explore the clinical and genetic features of two Chinese pedigrees affected with Autosomal dominant intellectual developmental disorder 49 (MRD49).Methods:Two MRD49 pedigrees which were admitted to the Children′s Hospital Affiliated to Shandong University respectively on January 28, 2021 and November 10, 2022 were selected as the study subjects. Clinical data of the two pedigrees were collected and analyzed. Genomic DNA was extracted from peripheral blood samples of the probands and their family members. The probands were subjected to mutational analysis by high-throughput sequencing. Candidate variants were validated using real-time fluorescence quantitative PCR (q-PCR) or Sanger sequencing and bioinformatic analysis. This study was approved by the Medical Ethics Committee of the Children′s Hospital Affiliated to Shandong University (No. SDFE-IRB/T-2022002).Results:Proband 1 had presented with language delay, motor retardation and intellectual disability, and his maternal grandmother, mother, aunt and cousin all had various degrees of intellectual disability. Sequencing results showed that proband 1 had deletion of exons 3 ~ 7 of the TRIP12 gene. q-PCR verification showed that his mother, aunt, maternal grandmother and cousin had all harbored the same deletion. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP1). Proband 2, who had mainly presented with language delay, motor retardation and intellectual disability, and was found to harbor a heterozygous c.3010C>T (p.Arg1004*) variant of the TRIP12 gene, which was verified to be de novo in origin. Based on the guidelines from the ACMG, the variant was classified as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:This study had diagnosed two MRD49 families through high-throughput sequencing. Above findings have enriched the phenotypic and mutational spectrum of MRD49 in China, which has also facilitated genetic counseling for the two pedigrees.

Background Environmental pollutants can affect N⁶-methyladenosine (m⁶A) level in the body, but the change of m⁶A level in kidney after being exposed to cadmium (Cd) and the molecular mechanism of renal injury need to be further studied. Objective To analyze the associations of m⁶A modification and methyltransferases/demethylases with microRNA-21 (miR-21) and transforming growth factor- β1 (TGF - β1) in kidney of rats exposed to Cd. Methods Twenty-four SPF male SD rats were divided into 4 groups, with 6 rats in each group, and were exposed to Cd by subcutaneous injection of 2.0, 1.0, and 0.5 mg·kg⁻¹ cadmium chloride (CdCl₂) and equal volume of normal saline for 2 weeks, 7 d a week, respectively. The levels of N-acetyl-β-D-glucosidase (UNAG) and albumin (UALB) in urine, and the levels of m⁶A methylation and TGF-β1 in kidney were detected by enzyme-linked immunosorbent assay (ELISA). The level of blood urea nitrogen (BUN) was measured by urease method. The levels of renal oxidative stress indicators such as malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were detected by total bile acid method, water-soluble tetrazolium asssay, and colorimetric method respectively. The relative levels of TGF-β1, methyltransferases, and demethylases in kidney were measured by reverse transcription-polymerase chain reaction. The expression of miR-21 in kidney was detected by fluorescent quantitative polymerase chain reaction. Results After 2 weeks of exposure to Cd, the body weights of rats in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups decreased, and the ratio of kidney/body weight and the levels of BUN, UNAG, and TGF-β1 mRNA and protein increased in the 2.0 mg·kg⁻¹ cadmium chloride group (P<0.05). The expression levels of m⁶A modification, methyltransferases METTL3, METTL14, Wilms’ tumor 1-associated protein (WTAP), and miR-21 were increased both in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups, with significant differences compared with the control group (P<0.05). The results of correlation analysis showed that the m⁶A modification level was negatively correlated with SOD (r=−0.4489, P<0.05) and GSH-Px (r=−0.4874, P<0.05), METTL3 was negatively correlated with MDA (r=−0.5158, P<0.05), while there was a positive correlation between FTO and GSH-Px (r=0.4802, P<0.05). In addition, miR-21 was positively correlated with METTL3 (r=0.7491), METTL14 (r=0.6157), and WTAP (r=0.6660) (P<0.05), TGF-β1 was positively correlated with METTL3 (r=0.5025, P<0.05) but negatively correlated with FTO (r=−0.5634, P<0.05) . Conclusion Cd can induce m⁶A methylation and up-regulation of METTL3, METTL14, WTAP, and miR-21 expression levels in rat kidney tissues, indicating that m⁶A and miR-21 may be associated with Cd-induced renal fibrosis.

Objective:To understand the incidence of post-traumatic stress disorder(PTSD) in fathers with premature infants and analyze the risk factors.Methods:Conveniently, the 203 fathers of premature infants in NICU of Affiliated Hospital of Jining Medical University of Shandong Province from May to August 2021 were selected as the subjects to fill in the general data questionnaire, Perinatal Post-traumatic stress disorder Questionnaire-Chinese edition (PPQ-C), Parents, Perception of Uncertainty Scale, Social Support Rating Scale, Simplified Coping Style Questionnaire. Logistic regression was used to analyze the risk factors of PTSD and establish a nomogram model. ROC curve was used to verify the discrimination ofthe model. Hosmer-Lemeshow goodness-of-fit test and Calibration Plot were used to verify the calibration.Results:PPQ-C total score of 203 fathers was 17.17 ± 8.77, 81 fathers, symptoms were positive and the incidence of PTSD was 39.90%(81/203). Logistic regression analysis showed college degree ( OR = 0.297, 95% CI 0.116 - 0.763, P<0.05), very low birth weight ( OR = 2.491, 95% CI 1.027 - 6.044, P<0.05), sense of disease uncertainty ( OR = 1.038, 95% CI 1.012 - 1.066, P<0.05), negative coping style ( OR = 1.871, 95% CI 1.127 - 3.108, P<0.05) were risk factors of PTSD in fathers with premature infants. The nomogram model was established basing on the results of the Logistic regression analysis, and the ROC curve proved (AUC = 0.751) the model having a good discrimination.The Hosmer-Lemeshow goodness-of-fit test ( P = 0.974) and the calibration plot demonstrated that the prediction values tends to coincide with the actual monitoring values. Conclusions:A higher incidence of PTSD was observed in fathers with hospitalized premature infant. It was related to education level, the infants′s birth weight, disease uncertainty and coping style. Therefore, the nurses should give information support and psychological guidance according to the individual situation of the infants′ father to reduce the incidence of PTSD.

Objective:To explore the research status, research focus and development trend of nursing human resource allocation in nursing homes in China.Methods:The article on nursing human resource allocation in nursing homes in China was systematically retrieved from China National Knowledge Infrastructure (CNKI), VIP and WanFang Data, and the retrieval time limit was from the establishment of the database to June 2022. CiteSpace was used to analyze the amount of articles, authors, institutions, funds and keywords, and draw a knowledge map.Results:A total of 890 articles were included. The number of articles and the support of national level funds were growing. The cooperation network among authors was extensive but not deep enough, and a core group of authors was not formed. A core journal group was formed. The research institute with the largest number of articles was the School of Nursing of Peking University. The article was mainly distributed in Beijing, Shanghai, Guangzhou and other first tier cities. The high-frequency keywords were "combination of medical care and nursing care" "supply" "demand" "talent training" and " care service for the aged", which could be clustered into 11 tags. The keyword cluster atlas showed that the combination of medical care and nursing care had always been the mainstream research direction in the field of nursing human resource allocation in nursing homes in China. "Qualitative research" "rural care for the aged " "care model for the aged " and "care industry for the aged " would become the emerging development trend in this field.Conclusions:The research on the nursing human resource allocation in nursing homes in China is growing. The research focuses on the combination of medical care and nursing care, talent training, care industry for the aged, and the supply and demand of care service for the aged. In the future, we should take the combination of medical care and nursing care as the leading mode, constantly improve our elderly care service system, build a nursing human resource allocation mode with Chinese characteristics in nursing homes, and promote the healthy aging development of society.