ObjectiveTo screen a set of short tandem repeat (STR) markers covering rat chromosomes 1-20 and the X chromosome, with 2-4 markers on each chromosome, and establish a dedicated marker panel for genetic contamination detection and strain identification of 5 commonly used inbred rat strains. MethodsSix 6-8 week-old rats (half male and half female) were selected from F344, BN, DA, Lewis, and PVG strain, respectively. Genomic DNA was extracted from rat tail tissues. A total of 61 STR markers were selected, including 27 from the national standard GB 14923—2022 and 34 from the literature. Singleplex STR PCR amplification combined with capillary electrophoresis was used for genotyping of the 5 inbred rat strains, and genotype data were analyzed using GeneMapper ID v3.2 software. Based on the STR genotyping results, genetic distances between strains were calculated with GenAlEx 6.51b2 software, and a phylogenetic tree of the inbred rat strains was constructed using MEGA7 software. ResultsAmong the 61 STR markers, non-specific amplification was observed for D15mit3, while no specific amplification product was obtained for D3wox7. LCA, AGT, and D5Hmgc2 exhibited no inter-strain polymorphism across the 5 inbred strains, while the remaining 56 markers exhibited inter-strain polymorphism. These 56 markers covered rat chromosomes 1-20 and the X chromosome (2-4 markers per chromosome). Among them, 42 markers could be used for genetic contamination detection in inbred rat strains. The amplified product lengths of D7wox14, D15rat123, and D20wox3 differed among the 5 strains, which could be applied for strain identification. The phylogenetic tree showed that the number of STR allelic differences between BN and F344 strains was 49, with a genetic distance of 1.775, both of which were the highest among all strain pairs. This indicated that the two strains exhibited the highest level of genetic differentiation and were the most distantly related. Among the 5 strains, F344, DA, and PVG clustered together, indicating their relatively close genetic distance, while BN and Lewis formed another cluster, indicating a relatively close genetic distance between them. The genetic relationships among the 5 strains were consistent with previous reports. ConclusionThis study successfully screened a set of STR markers for molecular genetic detection of 5 commonly used inbred rat strains, including F344, BN, DA, Lewis, and PVG, and established specific marker combinations for genetic contamination detection and strain identification, respectively.

Objective: To investigate the potential of conventional cranial computed tomography (CT) in assessing the early neurological deterioration(END), long-term prognosis, and traditional Chinese medicine (TCM) syndrome elements during the acute phase in patients with acute ischemic stroke (AIS). Methods: This study included 101 patients with AIS onset within 24 h in the Emergency Department of Fangshan Hospital, Beijing University of Chinese Medicine, from November 2019 to May 2021. To investigate the correlation between the relevant characteristics of the first conventional cranial CT in patients with AIS onset within 24 h and END, 90 d prognosis, and initial syndrome elements, the presence or absence of END, the 90 d prognosis (non-disabling outcome or functionally independent outcome), and the establishment of syndrome elements (internal fire, phlegm-dampness, blood stasis, qi deficiency, yin deficiency) were used as dependent variables and grouping criteria. Results: This study included 61 males and 40 females, with an age of (64.43±10.56) years. The time from onset to conventional cranial CT examination was 3.50 (1.50, 9.75) h. Among the patients, there were 70 cases (69.3%) of mild AIS, 30 cases (29.7%) of moderate AIS, and one case (1.0%) of severe AIS. Fifteen patients (14.9%) received intravenous thrombolysis. Among the 101 patients, six syndrome elements were observed within 24 h of onset: internal wind in 101 cases (100.0%), internal fire in 58 cases (57.4%), phlegm-dampness in 60 cases (59.4%), blood stasis in 67 cases (66.3%), qi deficiency in 39 cases (38.6%), and yin deficiency in 23 cases (22.8%). The incidence of END was higher in patients with lesions in the contralateral cerebral hemisphere to the affected limb (32.9%) than in those without such lesions (10.7%), showing a strong positive correlation with END occurrence (OR=4.082, P = 0.026). The incidence of END was higher in patients with lesions in the basal ganglia region (33.3%) and the carotid system blood supply area (32.8%) than in those without lesions in the basal ganglia region (15.8%) and the carotid system territory (14.7%), showing moderate positive correlations with END occurrence (OR=2.667, P =0.047; OR=2.836, P=0.044). The proportion of non-disabling outcomes was lower among patients with white matter degeneration (30.8%) and lesions in the contralateral cerebral hemisphere to the affected limb (52.1%) than in those without white matter degeneration (63.6%) and without such lesions in the contralateral cerebral hemisphere to the affected limb (78.6%), both showing strong negative correlations with the occurrence of non-disabling outcomes (OR=0.254, P=0.034; OR=0.296, P=0.015). Similarly, the proportion of functionally independent outcomes was lower among individuals with white matter degeneration (30.8%) and lesions in the contralateral cerebral hemisphere to the affected limb (64.4%) than in those without white matter degeneration (77.3%) and without such lesions in the contralateral cerebral hemisphere to the affected limb (89.3%), both also showing strong negative correlations with the occurrence of functionally independent outcomes (OR=0.131, P=0.001; OR=0.217, P=0.014). The incidence rates of internal fire, blood stasis, and yin deficiency syndrome elements were 66.7%, 73.0%, and 30.2%, respectively, among patients with lesions in the basal ganglia region, compared to 42.1%, 55.3%, and 10.5% among those without lesions in this region. The presence of lesions in the basal ganglia region showed moderate to strong positive correlations with internal fire and yin deficiency syndrome elements (OR=2.750, P=0.016; OR=3.670, P=0.028). Patients with lesions in the centrum semiovale and corona radiata regions (66.7%) had a higher incidence of qi deficiency than those without lesions in this region (33.7%), showing a strong positive correlation with the occurrence of qi deficiency (OR=3.931, P=0.022). No CT characteristics were found to be correlated with phlegm-dampness syndrome elements. Conclusion The first cranial CT in patients with AIS has potential application value in predicting disease progression, assessing prognosis, and diagnosing syndromes, which can provide physicians with diagnostic and treatment decisions to improve the long-term prognosis of patients with AIS.

Objective:To explore the therapeutic effects of Jiugong abdominal meridian massage combined with Zhuang medicine line acupoint moxibustion in patients after radical resection of colorectal cancer.Methods:A convenience sampling method was used to select 200 patients who underwent radical resection of colorectal cancer at Huzhou First People's Hospital from January 2022 to December 2023. Patients were randomly divided into three groups using a random number table: the control group ( n=62), the Zhuang medicine line acupoint moxibustion group ( n=66), and the combined group ( n=72). The control group received conventional interventions, the Zhuang medicine line acupoint moxibustion group received Zhuang medicine line acupoint moxibustion in addition to the conventional intervention, and the combined group received Jiugong abdominal meridian massage in addition to the Zhuang medicine line acupoint moxibustion. Gastrointestinal function recovery and pain scores were compared among the three groups after the intervention. Results:After intervention, there were statistically significant differences in the recovery time of bowel sounds, time to tolerate solid food by mouth, time to first bowel movement, gastric body motility frequency, total gastrointestinal output rate, gastric emptying rate, and pain scores among the three groups ( P<0.05) . Conclusions:Jiugong abdominal meridian massage combined with Zhuang medicine line acupoint moxibustion intervention can effectively improve the gastrointestinal function indicators and reduce postoperative pain in patients after radical resection of colorectal cancer.

Objective:To investigate the intervention effect and active mechanism of traditional Chinese medicine Marsdenia tenacis-sima on colitis-associated colorectal cancer(CAC).Methods:High-resolution mass spectrometry and a precise mass number data-base were used to identify the chemical components of Marsdenia tenacissima,and network pharmacology methods were used to obtain the main active components and targets of Marsdenia tenacissima in the treatment of colorectal cancer(CRC)and analyze its mecha-nism of action against CRC.A mouse model of CAC was established,and then Marsdenia tenacissima was analyzed in terms of its effect on disease activity index(DAI),body mass,colorectal morphology,and serum levels of tumor necrosis factor-α(TNF-α)and interleu-kin-1β(IL-1β)in CAC mice.Furthermore,16S rDNA sequencing technique was used to assess the impact of Marsdenia tenacissima on the diversity of intestinal microbiota in mice.Results:A total of 50 components were identified by mass spectrometry,and 37 active components were identified by network pharmacology.Furthermore,313 targets related to CRC were identified.A total of 958 GO terms and 173 KEGG terms were obtained,among which inflammation,oxidative stress,and cancer pathways showed a relatively high de-gree of association with Marsdenia tenacissima.After modeling,the mice in each group had the manifestations such as diarrhea,bloody stool,prolapse of anus,and weight loss,and the model group had a significant increase in DAI score after each DSS cycle,while the Marsdenia tenacissima group had a significant reduction in DAI score of the model mice after the first,second,and third cycles(P=0.007,P<0.001,and P<0.001).In addition,compared with the model group,the Marsdenia tenacissima group had a significant in-crease in colorectal length,alleviation of intestinal swelling,a relatively intact structure of epithelial cells,a reduction in inflammatory cell infiltration,a significant increase in goblet cells,a reduction in atypical hyperplasia,and significant reductions in the levels of TNF-α and IL-6 in CAC mice(P<0.001 and 0.002).The Marsdenia tenacissima group had significant reductions in the abundance and diversity of intestinal microbiota,and compared with the model group,the Marsdenia tenacissima group had a significant reduction in the relative abundance of Bacteroides and significant increases in the relative abundances of Muribaculaceae and Clostridia-UCG-014(P=0.002 and 0.009).Conclusion:Marsdenia tenacissima exerts an intervention effect on colorectal cancer in CAC mice,possibly by regulating the pathways associated with inflammation and cancer and altering the diversity of intestinal microbiota.

Objective:To explore the therapeutic effects of Jiugong abdominal meridian massage combined with Zhuang medicine line acupoint moxibustion in patients after radical resection of colorectal cancer.Methods:A convenience sampling method was used to select 200 patients who underwent radical resection of colorectal cancer at Huzhou First People's Hospital from January 2022 to December 2023. Patients were randomly divided into three groups using a random number table: the control group ( n=62), the Zhuang medicine line acupoint moxibustion group ( n=66), and the combined group ( n=72). The control group received conventional interventions, the Zhuang medicine line acupoint moxibustion group received Zhuang medicine line acupoint moxibustion in addition to the conventional intervention, and the combined group received Jiugong abdominal meridian massage in addition to the Zhuang medicine line acupoint moxibustion. Gastrointestinal function recovery and pain scores were compared among the three groups after the intervention. Results:After intervention, there were statistically significant differences in the recovery time of bowel sounds, time to tolerate solid food by mouth, time to first bowel movement, gastric body motility frequency, total gastrointestinal output rate, gastric emptying rate, and pain scores among the three groups ( P<0.05) . Conclusions:Jiugong abdominal meridian massage combined with Zhuang medicine line acupoint moxibustion intervention can effectively improve the gastrointestinal function indicators and reduce postoperative pain in patients after radical resection of colorectal cancer.

OBJECTIVE To establish an HPLC method for determination of the related substances in papaverine hydrochloride. METHODS NanoChrom ChromCore 120 C8 column was used; the mobile phase A consisted of 3.4 g·L−1 potassium dihydrogen phosphate aqueous solution, adjust pH to 3.5 with phosphoric acid-acetonitrile(90∶10), the mobile phase B was methanol, with gradient elution at the flow rate of 0.8 mL·min−1; the detection wavelength was 238 nm; the column temperature was 50 ℃. RESULTS The minimum separation between the main component and each impurity was >1.5; Papaverine and its thirteen impurities showed a good linear relationship in the self-concentration range(r>0.999); and the average recoveries were 93.1%−101.2% with RSDs of 2.3%−8.1%. CONCLUSION The method is accurate, sensitive and reliable, which is suitable for the determination of related substances in papaverine hydrochloride.

Objective:To analyze the mutation characteristics of rpoB gene in rifampicin-resistant Brucella strains. Methods:DNA of 4 rifampicin-resistant Brucella strains (JSY-26, G-9, WSY-13 and AW-3) isolated from Xinjiang Uygur Autonomous Region was selected, rifampicin rpoB gene was amplified by PCR and its nucleotide sequence was sequenced. The rpoB gene sequences of rifampicin-resistant Brucella standard strain (RB51) and sensitive strain (ALT-8) were used as reference, the mutation sites and types of the rpoB gene inside and outside the rifampicin resistance determination region (RRDR) of the 4 rifampicin-resistant Brucella strains were analyzed by Mega 7.0 software. Results:Through sequence alignment, both JSY-26 and WSY-13 strains underwent a single base point mutation at the RRDR 1 576 bp of the rpoB gene, with the base changing from guanine (G) to adenine (A). The G-9 strain underwent a single base point mutation at the RRDR 1 606 bp of the rpoB gene, with the base changing from cytosine (C) to A. The AW-3 strain showed 5 mutations of 3 types outside rpoB gene RRDR at 2 536, 2 537, 2 626, 2 636 and 2 654 bp, namely 3 insertion mutations [thymine (T) insertion once and C insertion twice], 1 deletion mutation (C deletion), and 1 single base point mutation (from G to C mutation).Conclusion:The RRDR mutations in the rpoB gene of the rifampicin-resistant Brucella strains are mainly characterized by single base point mutations, while multiple insertion and deletion mutations occur outside the RRDR.

Hippo signaling pathway plays an important role in cell growth, proliferation, apoptosis, and stem cell regeneration. Dysregulation of this pathway has been linked to various diseases, such as cancer, heart disease, lung disease, kidney disease, liver disease, and immune dysfunction, as well as ocular diseases. Consequently, regulating the balance of Hippo signaling pathway may be an effective treatment for ocular diseases. This article reviews the mechanism of action and therapeutic potential of Hippo signaling pathway in ocular diseases, and discusses the potential therapeutic targets of the Hippo signaling pathway, to identify new targets and provide new research ideas for treating ocular diseases.

Metabolic diseases refer to metabolic disorders caused by abnormal material metabolism or energy metabolism of the body such as diabetes,hyperlipidemia,hyperuricemia and liver disease.In recent years,with the change of national diet structure,its incidence has increased year by year,which seriously endangers national physical and mental health.In modern medicine,long-term treatment of metabolic diseases is often used in the treatment of chemical drugs,which is easy to produce drug resistance and liver and kidney damage.Tannin is a class of polyphenolic compounds composed of glucose,gallic acid,flavanols,etc.,which has antibacterial,anti-tumor,anti-oxidation,hypoglycemic,antiviral,anti-inflammatory and analgesic effects,and has a clear easing effect on metabolic diseases.Tannin in the treatment of metabolic diseases has the advantages of less toxic side effects,significant curative effect,low cost,easy to popularize,etc.,which provides a better choice for the treatment of metabolic diseases.

ObjectiveTo establish a set of single nucleotide polymorphisms (SNP) detection protocol for inbred rats based on multiplex PCR-ligase detection reaction (LDR). MethodsA total of 40 rats SNP sites were selected on chromosomes 1-20 and X of rats among 5 inbred strains of rats, and the 40 SNP sites were randomly divided into four groups. A genetic detection protocol for 4 groups of SNP in inbred rats based on multiplex PCR-LDR technology was constructed. 9 commonly used rat strains from two other domestic rat suppliers were detected by this protocol. Finally, the feasibility of this protocol was verified by comparing the amplification effects of different DNA polymerases by a third-party laboratory. ResultsWhen using the constructed SNP detection protocol for inbred rats to test 5 rat strains, all sites in each sample obtained good amplification results. The 9 commonly used rat strains from two other rat suppliers in china were also well amplified by this SNP detection protocol, and 40 SNPs were homozygous in each Inbred strain. The results of detection of the same rat DNA samples with three different DNA polymerases showed that the Multiplex PCR Kit, AmpliTaq Gold 360 DNA polymerase and Platinum II Taq hot start DNA polymerase had electrophoretic peaks of amplification products at all SNP sites in groups 1 to 3, and Platinum II Taq hot start DNA polymerase had one less electrophoretic peak of the amplification products at the SNP sites in group 4. In addition, inter-laboratory comparisons showed consistent results for the same amplification system. ConclusionBased on multiplex PCR-LDR technology, this study successfully established a SNP detection protocol for rats covering all autosomes and X chromosomes with the excellent stability and repeatability.