Objective: To evaluate the clinical efficacy of digitally guided precision crown lengthening in secondary aesthetic rehabilitation cases, and to provide a clinical reference for digitally guided crown lengthening procedures and secondary aesthetic restorations. Methods: We present a case of a patient with tetracycline-stained teeth, partial detachment of anterior resin veneers, and gingival margin discrepancies. The patient underwent digitally guided precision crown lengthening followed by secondary aesthetic rehabilitation. Multimodal data, including intraoral, facial, and CBCT scans, were integrated to construct a four-dimensional virtual patient model (incorporating teeth, face, bone, and occlusion) for surgical planning and 3D-printed guide fabrication. Secondary aesthetic restoration was performed after achieving stable post-surgical outcomes. Based on this case, we conducted a detailed analysis and reviewed relevant literature on crown lengthening in secondary aesthetic rehabilitation. Results: The gingival contour of the anterior teeth exhibited significant improvement, with enhanced symmetry and stable gingival margin positioning that closely matched the preoperative design. The crown lengthening procedure demonstrated high precision, and the final outcome was aesthetic and functional. Literature review indicated that secondary restorations frequently present challenges such as gingival contour discrepancies and inflammation. Aesthetic crown lengthening in the anterior region should optimize both soft and hard tissue morphology to meet aesthetic standards, with digital technology improving procedural accuracy. Conclusion Precision crown lengthening effectively addresses gingival margin discrepancies in secondary aesthetic rehabilitation, ensuring stable gingival positioning and superior aesthetic outcomes. This approach is particularly suitable for cases with high aesthetic demands.

Tryptophan metabolism plays an important role in immunometabolism in sepsis,and the expression of indoleamine 2,3-dioxygenase 1(IDO1),the key enzyme of tryptophan metabolism,is up-regulated during sepsis.This study was designed to construct IDO1 gene knockout THP-1 cell line using CRISPR/Cas9,and to provide a cell model for studying the role of IDO1 in macrophage function.Three gRNAs were designed for the IDO1 gene,and inserted into YKO-Lentiviral gRNA plasmid respectively to construct IDO1-gRNA recombinant plasmid,which then confirmed by restriction enzyme digestion and sequencing,and packaged as Lenti-IDO1-gRNA lentivirus.THP-1 cells were infected with Cas9 lentivirus to construct a THP-1 Cas9 cell line stably expressing Cas9 protein.Then THP-1 Cas9 cells were infected with the Lenti-IDO1-gRNA lentivirus for IDO1 gene knockout,and the monoclonal cell lines were screened by limited dilution method.The efficiency of IDO1 knockout was identified by T7E1 digestion test,PCR product sequencing and Western blotting.The proliferative activity of THP-1 cells was detected by CCK8 assay;the expression of CD11b,CD68 and CD 14 in THP-1 macrophages were detected by flow cytometry;and the phagocytic function of THP-1 macrophages was detected by neutral phagocytosis test.T7E1 restriction enzyme digestion results showed that all the three gRNAs could effectively edit IDO1 gene,and gRNA2 had the highest editing efficiency.Sequencing of PCR products showed that IDO1 gene was mutated in three THP-1 IDO 1-KO monoclonal cell lines,and Western blot result showed that IDO1 protein was not expressed,suggesting THP-1 IDO1-KO cells were successfully constructed.CCK-8 proliferation assay showed that IDO1 gene knockout could inhibit the proliferation of THP-1 cells.Flow cytometry showed that IDO1 gene knockout down-regulated the expression of CD1 1b and CD68 in THP-1 macrophages,while up-regulated the expression of CD 14.The results of neutral red phagocytosis test showed that IDO1 gene knockout promoted the phagocytosis of macrophages.In conclusion,the IDO1 gene knockout THP-1 cell line has successfully constructed by CRISPR-Cas9,and IDO1 plays an important role in regulating the proliferation activity and differentiation of THP-1 cells,as well as the phagocytic function of THP-1 macrophages.

Objective:To understand the infection status and molecular types of rhinovirus (RV) among cases of Acute Respiratory Infections (ARIs) in Luohe City, Henan Province, from 2017 to 2022.Methods:From October 2017 to June 2022, clinical and epidemiological data were collected from 2 270 cases of ARIs at Luohe Central Hospital in Henan Province. Throat swab specimens were obtained from these cases. Real-time quantitative polymerase chain reaction (qPCR) was used to screen for RV-positive specimens. Subsequently, the positive samples were subjected to nested reverse transcription polymerase chain reaction (nested RT-PCR) to amplify the full-length VP1 region. Using the MEGA software, along with 169 RV reference strains recommended by the International Committee on Taxonomy of Viruses, a phylogenetic tree was constructed to determine RV types.Results:Among the 2 270 cases of ARIs, there were 1 283 male cases (56.52%). The median age ( Q 1, Q 3) was 3 (1, 6) years, with the population under 5 years old accounting for 68.59% (1 557/2 270). RV was detected in 137 cases (6.04%), of which 68 cases (49.64%) showed co-detection with other viruses, with the most common being co-detection with enterovirus, accounting for 14.60% (20/137). The RV detection rates in the age groups of 0-4 years, 5-14 years, 15-59 years, and≥60 years were 6.42% (100/1 557), 4.69% (21/448), 3.80% (6/158), and 9.35% (10/107), respectively, with no statistically significant differences ( χ2=5.310, P=0.150). The overall detection rates of RV before (2017-2019) and during (2020-2022) the COVID-19 pandemic showed no statistically significant difference ( χ2=1.823, P=0.177). A total of 109 VP1 sequences were obtained, including 62 types. Among them, RV-A, RV-B, and RV-C had 42, 3, and 17 types respectively. Conclusion:RV is one of the predominant pathogens in ARIs cases in Luohe City, Henan Province, from 2017 to 2022. Multiple types of RV co-circulate without any apparent dominant type.

Objective To investigate the effect of ultrasound-guided bilateral retrolaminar block(RLB)on postoperative analgesia and myocardial protection in the patients with heart valve replacement under car-diopulmonary bypass(CPB).Methods A total of 70 patients with scheduled valve replacement under CPB in this hospital from May 2022 to March 2023 were selected as the study subjects and divided into the simple general anesthesia group(group A)and ultrasound-guided bilateral RLB combined general anesthesia group(group B)by the random number table method,35 cases in each group.There was no special treatment in the group A;before anesthesia induction,the group B underwent bilateral RLB under ultrasound guidance through the laminar level of the fourth thoracic vertebra.All patients used the patient-controlled intravenous analgesia(PCIA)pump after surgery.The mean arterial pressure(MAP)and heart rate(HR)of the two groups were recorded before anesthesia induction(T0),during intubation(T1),during sternal incision(T2),before CPB(T3),at 10 min after CPB stop(T4)and at 5 min after surgery(T5).The time of first awakening,extubation time and scores of the Numeric Pain Scale(NRS)in the resting and coughing states after extubation were re-corded in the two groups.The dose of intraoperative sufentanil and the oxycodone dose for remedy analgesia after extubation were recorded in the two groups.The levels of malondialdehyde(MDA),superoxide dis-mutase(SOD),cardiac troponin Ⅰ(cTnⅠ)and creatine kinase isoenzyme(CK-MB)were detected by ELISA at T0,30 min after CPB beginning(T6),30 min after CPB end(T7),postoperative 24 h(T8)and 48 h(T9).The occurrence situation of adverse events such as nausea and vomiting,puncture site infection,local anesthetic poisoning and puncture injury were recorded in the two groups.Results Compared with the group A,MAP and HR at T1,T2 and T3 in the group B were significantly decreased.The time of first awakening and extuba-tion after surgery were advanced.After extubation,the NRS score at rest and cough was significantly reduced;the dosage of sufentanil and oxycodone was reduced;the levels of MDA,cTnⅠ and CK-MB at T6,T7 and T8 were significantly decreased,the SOD level was significantly increased,and the differences were statistically significant(P＜0.05).There were no adverse events occurrence such as nausea and vomiting,puncture site in-fection,local anesthetic poisoning and puncture injury in both groups.Conclusion Ultrasound-guided bilateral RLB could stabilize the intraoperative hemodynamics and provide perfect analgesia in the patients with CPB heart valve replacement,meanwhile reduce the stress response and play certain role in myocardial protection.

Objective:To understand the infection status and molecular types of rhinovirus (RV) among cases of Acute Respiratory Infections (ARIs) in Luohe City, Henan Province, from 2017 to 2022.Methods:From October 2017 to June 2022, clinical and epidemiological data were collected from 2 270 cases of ARIs at Luohe Central Hospital in Henan Province. Throat swab specimens were obtained from these cases. Real-time quantitative polymerase chain reaction (qPCR) was used to screen for RV-positive specimens. Subsequently, the positive samples were subjected to nested reverse transcription polymerase chain reaction (nested RT-PCR) to amplify the full-length VP1 region. Using the MEGA software, along with 169 RV reference strains recommended by the International Committee on Taxonomy of Viruses, a phylogenetic tree was constructed to determine RV types.Results:Among the 2 270 cases of ARIs, there were 1 283 male cases (56.52%). The median age ( Q 1, Q 3) was 3 (1, 6) years, with the population under 5 years old accounting for 68.59% (1 557/2 270). RV was detected in 137 cases (6.04%), of which 68 cases (49.64%) showed co-detection with other viruses, with the most common being co-detection with enterovirus, accounting for 14.60% (20/137). The RV detection rates in the age groups of 0-4 years, 5-14 years, 15-59 years, and≥60 years were 6.42% (100/1 557), 4.69% (21/448), 3.80% (6/158), and 9.35% (10/107), respectively, with no statistically significant differences ( χ2=5.310, P=0.150). The overall detection rates of RV before (2017-2019) and during (2020-2022) the COVID-19 pandemic showed no statistically significant difference ( χ2=1.823, P=0.177). A total of 109 VP1 sequences were obtained, including 62 types. Among them, RV-A, RV-B, and RV-C had 42, 3, and 17 types respectively. Conclusion:RV is one of the predominant pathogens in ARIs cases in Luohe City, Henan Province, from 2017 to 2022. Multiple types of RV co-circulate without any apparent dominant type.

OBJECTIVE To enrich the polymethoxyflavonoid-rich fraction from Citri Reticulatae Pericarpium using D101 macro-porous resin,and further to analyse the chemical constituents using High-Performance Liquid Chromatography Quadrupole-Time-of-Flight Mass Spectrometry(HPLC-Q-TOF-MS).METHODS Citri Reticulatae Pericarpium extract was adsorbed on D101 macro-porous resin and then eluted with different concentrations of ethanol to enrich the polymethoxyflavonoid-rich fraction;the content of the major flavonoid components was determined by high-performance liquid chromatography(HPLC);and the components were further an-alysed by HPLC-Q-TOF-MS.RESULTS The established HPLC method for the simultaneous determination of seven flavonoids in Citri Reticulatae Pericarpium extract was accurate and reliable.The contents of compounds such as nobiletin and tangeritin were signifi-cantly increased in the polymethoxyflavonoid-rich fraction.Using high resolution mass spectrometry(HRMS),70 and 60 compounds were identified from the Citri Reticulatae Pericarpium extract and the polymethoxyflavonoid-rich fraction,respectively,with 53 polyme-thoxyflavonoids being detected in these two extracts.CONCLUSION The results of this study provide an experimental basis for fur-ther identification of the pharmacological substance basis of the polymethoxyflavonoids in Citri Reticulatae Pericarpium and the estab-lishment of quality control methods.

In recent years, the prevalence of gestational diabetes mellitus (GDM) has significantly increased, leading to various adverse outcomes for mothers and their offspring. Early diagnosis and good blood glucose control can significantly improve pregnancy outcomes. Currently, the diagnosis of GDM mainly relies on the oral glucose tolerance test (OGTT), but there are many limitations in clinical application. The new biological marker glycosylated CD59 is a glycosylation product of complement regulatory proteins that can stably exist in body fluids and is linearly related to blood glucose levels, which has the advantages of simple testing and good reproducibility, and has been developed into a stable reagent kit. It has shown good value in the early diagnosis, standard diagnosis, and prediction of pregnancy outcomes of GDM.

Objective To investigate the impact of the Shared Medical Appointment (SMA) model on blood glucose levels and self-management behaviors in patients with gestational diabetes mellitus (GDM). Methods A total of 87 pregnant women diagnosed with GDM at the Obstetrics and Gynecology and Endocrinology Outpatient Departments of the Affiliated Suqian Hospital of Xuzhou Medical University from October 2021 to October 2022 were enrolled and divided into treatment group (n=44) and control group (n=43). The treatment group received SMA-based GDM management, while the control group underwent routine outpatient follow-up. Baseline data, flash glucose monitoring (FGM)-related indicators before and after treatment, and self-management behavior scale scores were compared between the two groups. Results No statistically significant differences were observed between the two groups in age, body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FBG), blood urea nitrogen (BUN), creatinine (Cr), uric acid (UA), triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and glycated hemoglobin (HbA1c) (P>0.05). After treatment, the time in range (TIR) of glucose was higher in the treatment group than that in the control group, while mean amplitude of glycemic excursions (MAGE), standard deviation of blood glucose (SDBG), coefficient of variation (CV), and mean blood glucose (MBG) were lower in the treatment group (P < 0.05). Within the treatment group, post-treatment TIR was higher, and MAGE, CV, SDBG, and MBG were lower than pre-treatment levels (P < 0.05). In the control group, post-treatment TIR was higher, and SDBG and MBG were lower than pre-treatment levels (P < 0.05). After treatment, the total score and scores for dietary management, exercise management, medication management, and blood glucose monitoring management in the self-management behavior scale were significantly higher in the treatment group than in the control group (P < 0.05). In the treatment group, post-treatment scores for all self-management behaviors were higher than pre-treatment levels, while post-treatment total scores and scores for medication management and blood glucose monitoring management in the control group were higher than pre-treatment levels (P < 0.05). Conclusion SMA management model can comprehensively improve blood glucose levels and enhance self-management behavior capabilities in GDM patients.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.