ObjectiveThis paper aims to investigate and evaluate the staged efficacy and safety of the representative empirical prescription of the “Zhu Ke Jiao” theory， Qijia Rougan prescription， combined with entecavir in the treatment of hepatic fibrosis in chronic hepatitis B. MethodsA multicenter randomized controlled clinical study was conducted， and 101 patients diagnosed with chronic hepatitis B-related hepatic fibrosis （CHB-HF） who met the diagnosis and inclusion criteria were randomly assigned to an observation group （Qijia Rougan prescription + entecavir） and a control group （entecavir）. The treatment duration was 24 weeks. Liver stiffness measurement （LSM）， fibrosis-4 index （FIB-4）， portal vein diameter， hepatitis B serology， biochemical indicators， hepatic fibrosis markers in serum ［hyaluronic acid （HA）， laminin （LN）， procollagen Ⅲ peptide （PⅢP）， and type Ⅳ collagen （Ⅳ-C）］， and traditional Chinese medicine syndrome scores were used as efficacy evaluation indicators. Efficacy assessments and explorations of different staged subgroups of Qijia Rougan prescription were conducted according to LSM values based on the Metavir pathological staging standard. ResultsA total of 98 cases were included for statistical analysis， with 49 cases in the observation group and 49 in the control group. The general data of the patients in both groups were comparable. Compared with the same group before treatment， the observation group showed a significant reduction in LSM and FIB-4 （P<0.01）， as well as notable improvements in LN， Ⅳ-C， and various TCM syndrome scores （P<0.05， P<0.01）. When compared to the control group after treatment， the observation group demonstrated significant improvements in LSM， FIB-4， and various TCM syndrome score indicators （P<0.05， P<0.01）， indicating that the observation group performed better than the control group. Subgroup analysis of the regression of hepatic fibrosis stages showed that compared to the same group before treatment， the observation group had better improvement in regression of stages F2 and F3 （P<0.05）. When compared to the control group after treatment， the observation group exhibited superior improvement in regression of stage F3 （P<0.05）. No adverse events occurred in either group during the treatment period. ConclusionCompared with entecavir alone， the combination of Qijia Rougan prescription and entecavir significantly improves the degree of hepatic fibrosis and clinical TCM symptoms in patients. The optimal intervention period is primarily during stage F3， which is a potential “interception” point of the “Zhu Ke Jiao” theory.

Objective To obtain the recombinant Argonaute protein(TtAgo)from Thermus thermophilus using gene cloning and pro-tein purification techniques and detect its endonuclease activity against the KRAS 12D mutation in vitro,thereby to provide experimental evidence for its subsequent application in the detection of gene mutations.Methods The TtAgo gene sequence was amplified from the genome of Thermus thermophilus by the PCR technology.Then,the amplified sequence was inserted into the prokaryotic expression vec-tor pET-28a to construct the recombinant vector pET-28a-TtAgo.After the recombinant TtAgo protein was induced to be expressed,it was purified sequentially by the nickel ion affinity chromatography(Ni-NTA),heparin affinity chromatography,and size-exclusion chromatography.The high-specificity nuclease activity of the recombinant TtAgo protein mediated by a short DNA guide strand(gDNA)was verified in vitro.Results The prokaryotic expression vector pET-28a-TtAgo was successfully constructed by the gene cloning tech-nique,and the soluble expression of the recombinant TtAgo protein was achieved.With the help of protein purification technology,the high-purity recombinant TtAgo protein was obtained.Under the mediation of gDNA,the recombinant TtAgo protein could specifically cleave wild-type single-stranded DNA(ssDNA),double-stranded DNA(dsDNA),and RNA molecules,while had no cleavage activi-ty for tumor KRAS 12D mutant molecules.Conclusion The high-purity recombinant TtAgo protein with nuclease activity is successful-ly obtained,and its targeted cleavage activity towards the KRAS 12D mutation site is detected in vitro.

Objective To investigate the status of oral frailty(OF)in patients who underwent chemotherapy for lung cancer,identify key factors influencing OF,and develop a risk prediction model.Methods Using convenience sampling,431 lung cancer inpatient were recruited from three Tier-IIIA hospitals in Jiangsu Province between September and November 2024 as the training cohort.The patients were divided into OF and non-OF groups.Relevant data were compared between the two groups.Multifactorial logistic regression analysis was performed to determine factors that associated with OF,and a risk prediction model was created accordingly.Receiver operating characteristic(ROC)curve analysis was used to predict model performance.In December 2024,additional 185 patients from one other Tier-IIIA hospitals were recruited to validate the developed model.Results The prevalence of OF among lung-cancer patients undergoing chemotherapy was 58.93%.Following listed items were identified as the risk factors of OF(all P<0.05):older in age(OR=3.420),poor education(OR=0.030),brain metastasis(OR=7.880),high nutritional risk screening 2002 score(OR=1.550),elevated C-reactive protein(OR=1.100),and elevated lactate dehydrogenase(OR=1.010).ROC area under the curve(AUC)of the model was 0.860(95%CI:0.830-0.900)in modelling cohort and 0.840(95%CI:0.780-0.900)in validation cohort.Hosmer-Lemeshow goodness-of-fit test yielded χ 2=4.870,P=0.770 for the training set and χ 2=2.770,P=0.950 for the validation set.Conclusion The risk prediction model for OF developed in this study demonstrates a good predictive performance and can facilitate early identification of high-risk patients,thereby providing a scientific basis for clinical interventions.

OBJECTIVE T o understand the resistance capability of berberine combined with levofloxacin in vitro to methicillin-resistant Staphylococcus aureus(MRSA).METHODS Totally 100 strains of MRSA were isolated from Shanghai Eighth People's Hospital from Jul.2023 to Jun.2024,the minimum inhibitory concentrations(MICs)of berberine and levofloxacin against the strains were determined by microbroth dilution method.The anti-bacterial effect of the combined use of the two drugs on MRSA was observed.The effects of the single and com-bined use of berberine and levofloxacin on sterilization of MRSA standard strain USA 300-LAC and their influence on cellular morphology were observed.The influence of berberine on mRNA levels of drug resistance genes and ef-flux pump-related genes in the standard strain USA 300-LAC and clinical strains was determined by means of real-time fluorescent quantitative PCR.RESULTS Among the 100 clinical isolates of MRSA,most of them were isola-ted from respiratory tract specimens of respiratory medicine department and geriatrics department,and 75.00％of the strains were resistant to levofloxacin.The MICs of both berberine and levofloxacin against the standard strain USA300-LAC were 128 μg/ml;the MICs against the clinical MRSA isolates were 64 to 1024 μg/ml and 0.125 to 256 μg/ml,respectively.Berberine combined with levofloxacin showed additive effect on 17(44.74％)strains of MRSA.The combined use of the two drugs showed the most remarkable in vitro antibacterial effect.The cellular structures of the strains were unclear and the contents of bacterial bodies increased when berberine and levofloxacin were used in combination.Berberine showed various degree of inhibitory effect on gyrA,gyrB,grlA and nor A genes.CONCLUSIONS Berberine is capable of resisting the MRSA and shows synergistic or additive effect with levofloxacin.The combined use of the two drugs may reduce the MIC against levofloxacin,destroy the morpholog-ical structure,and inhibit the expression of gyrA,gyrB,grlA and nor A genes.

Objective To establish a gas chromatography-mass spectrometry method for simultaneous determination of the contents of volatile components muscone,menthol,borneol,isoborneol,camphor,and carvone in Wuli Huichun pills,and to evaluate the quality of Wuli Huichun pills based on authenticity,effectiveness and safety.Methods The ethyl acetate extract was analyzed by HP-5MS capillary column(30 m×0.25 mm,0.25 μm)with temperature programming.The injection port temperature was set at 230℃,and the split ratio was set at 10∶1.The carrier gas was high-purity helium gas.The injection port was in constant flow mode,with a carrier gas flow rate of 1.0 mL·min-1.The detector was a mass spectrometer with a triple quadrupole in series.The ion source was an electron bombardment source(EI)with a temperature of 250℃.The collision gas was argon.The temperature of the mass spectrometry transmission interface was 250℃.Scanning mode was SIM mode,with a solvent delay of 2 minutes.Results The linear relationships of the six components were good(r>0.999 0),the average recoveries were 93.57%-97.96%,and the RSD was 0.73%-2.5%.Conclusion The method is simple,accurate,and efficient,and can be used for the quality evaluation of Wuli Huichun pills.

Objective:To adapt the Mindful Breastfeeding Scale (MINDF-BFS) into Chinese and assess its reliability and validity among breastfeeding mothers in China.Methods:Following the Beaton cross-cultural adaptation guideline, the original scale was translated, back-translated, discussed by experts, pre-tested, culturally adapted, and revised to develop the Chinese version of the MINDF-BFS. A convenience sampling method was used to select 305 postpartum women from Guangzhou Women and Children's Medical Center, Guangzhou Medical University, who visited between March and June 2024, as the study participants. The reliability and validity of the Chinese version of MINDF-BFS were evaluated.Results:The Chinese version of MINDF-BFS consisted of nine items, with the item-level content validity index ranged from 0.900 to 1.000, and the average scale-level content validity index was 0.990. Exploratory factor analysis extracted one common factor, with a variance contribution of 73.290%. Confirmatory factor analysis showed a good model fit. The Cronbach's α coefficient for the scale was 0.923, the split-half reliability coefficient was 0.915, and the test-retest reliability coefficient was 0.926.Conclusions:The Chinese version of MINDF-BFS has good psychometric properties and is suitable for assessing the mindful breastfeeding levels of Chinese postpartum women.

Objective To obtain the recombinant Argonaute protein(TtAgo)from Thermus thermophilus using gene cloning and pro-tein purification techniques and detect its endonuclease activity against the KRAS 12D mutation in vitro,thereby to provide experimental evidence for its subsequent application in the detection of gene mutations.Methods The TtAgo gene sequence was amplified from the genome of Thermus thermophilus by the PCR technology.Then,the amplified sequence was inserted into the prokaryotic expression vec-tor pET-28a to construct the recombinant vector pET-28a-TtAgo.After the recombinant TtAgo protein was induced to be expressed,it was purified sequentially by the nickel ion affinity chromatography(Ni-NTA),heparin affinity chromatography,and size-exclusion chromatography.The high-specificity nuclease activity of the recombinant TtAgo protein mediated by a short DNA guide strand(gDNA)was verified in vitro.Results The prokaryotic expression vector pET-28a-TtAgo was successfully constructed by the gene cloning tech-nique,and the soluble expression of the recombinant TtAgo protein was achieved.With the help of protein purification technology,the high-purity recombinant TtAgo protein was obtained.Under the mediation of gDNA,the recombinant TtAgo protein could specifically cleave wild-type single-stranded DNA(ssDNA),double-stranded DNA(dsDNA),and RNA molecules,while had no cleavage activi-ty for tumor KRAS 12D mutant molecules.Conclusion The high-purity recombinant TtAgo protein with nuclease activity is successful-ly obtained,and its targeted cleavage activity towards the KRAS 12D mutation site is detected in vitro.

Objective To investigate the status of oral frailty(OF)in patients who underwent chemotherapy for lung cancer,identify key factors influencing OF,and develop a risk prediction model.Methods Using convenience sampling,431 lung cancer inpatient were recruited from three Tier-IIIA hospitals in Jiangsu Province between September and November 2024 as the training cohort.The patients were divided into OF and non-OF groups.Relevant data were compared between the two groups.Multifactorial logistic regression analysis was performed to determine factors that associated with OF,and a risk prediction model was created accordingly.Receiver operating characteristic(ROC)curve analysis was used to predict model performance.In December 2024,additional 185 patients from one other Tier-IIIA hospitals were recruited to validate the developed model.Results The prevalence of OF among lung-cancer patients undergoing chemotherapy was 58.93%.Following listed items were identified as the risk factors of OF(all P<0.05):older in age(OR=3.420),poor education(OR=0.030),brain metastasis(OR=7.880),high nutritional risk screening 2002 score(OR=1.550),elevated C-reactive protein(OR=1.100),and elevated lactate dehydrogenase(OR=1.010).ROC area under the curve(AUC)of the model was 0.860(95%CI:0.830-0.900)in modelling cohort and 0.840(95%CI:0.780-0.900)in validation cohort.Hosmer-Lemeshow goodness-of-fit test yielded χ 2=4.870,P=0.770 for the training set and χ 2=2.770,P=0.950 for the validation set.Conclusion The risk prediction model for OF developed in this study demonstrates a good predictive performance and can facilitate early identification of high-risk patients,thereby providing a scientific basis for clinical interventions.

To investigate the expression level of costimulatory molecule B7-H3 in the tumor tissues and the level of soluble costimulatory molecule B7-H3 (sB7-H3) in the serum of patients with colorectal cancer (CRC), so as to evaluate the clinical value of sB7-H3 in auxiliary diagnosis of CRC. A cross-sectional study design was adopted. A total of 232 CRC patients, 87 patients with benign colorectal diseases, and 59 healthy subjects who were treated in Shanghai Eighth People′s Hospital from January 2020 to December 2022 were selected. The levels of sB7-H3, CEA, CA199, CA724 and CA50 in the serum were detected. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of sB7-H3 and the above-mentioned tumor markers for colorectal cancer (CRC). The expression levels of B7-H3 in CRC tissues and benign colorectal disease tissues were detected by immunohistochemistry. The relationship between the levels of sB7-H3 and clinicopathological features was analyzed statistically. The results showed that compared with the benign disease group or the healthy control respectively, the serum levels of sB7-H3, CEA, CA199, CA724 and CA50 in the CRC group were significantly increased, and the differences were statistically significant ( P<0.05). In the CRC group, the serum levels of sB7-H3 showed a weak positive correlation with CA50, CEA and CA724 (the r values were 0.220, 0.217 and 0.182 respectively; the P values were 0.005,<0.001 and 0.024 respectively), and there was no significant correlation with CA199 (the r value was 0.162; the P value were 0.051). The areas under the curve (AUC) of sB7-H3, CEA, CA199, CA724 and CA50 for diagnosing CRC were 0.862, 0.774, 0.646, 0.677 and 0.644 respectively, and the cut-off values were 20.67 ng/ml, 10.74 U/ml, 3.17 ng/ml, 3.16 U/ml, and 22.55 U/ml, respectively. Taking 20.67 ng/ml as the cut-off value, the positive rate of sB7-H3 in CRC was 62.9%, which was significantly higher than that in patients with benign colorectal diseases (35.6%) and the healthy control group (10%) ( χ2=81.995, P<0.001; χ2=103.56, P<0.001). The positive rates of sB7-H3 and CEA in patients with pathological stages Ⅲ and Ⅳ were significantly higher than those in patients with stages Ⅰ and Ⅱ ( χ2=82.876, P<0.001; χ2=22.617, P<0.001). The positive rate of sB7-H3 in patients with pathological stages Ⅰ and Ⅱ was 56.2%, which was significantly higher than that of CEA (38%) ( χ2=50.378, P<0.001). Immunohistochemistry showed that B7-H3 positive staining was mainly distributed in the cytoplasm. The positive expression rate of B7-H3 in CRC (75.8%) was significantly higher than that in benign colorectal diseases (15.4%) ( χ2=16.133, P<0.001). The serum level of sB7-H3 in CRC patients was positively correlated with the expression level of B7-H3 in tumor tissues ( r=0.766, P<0.001). The serum level of sB7-H3 was significantly correlated with distant metastasis and pathological stage of CRC ( W=899, P=0.002; H=10.465, P=0.015). In conclusion, serum level of sB7-H3 may have certain clinical value in the auxiliary diagnosis of CRC.

Objective:To investigate the iodine nutrition status of children aged 8 - 10 and pregnant women in Xining City, Qinghai Province.Methods:From 2019 to 2021, a stratified cluster sampling method was used to divide 7 counties (districts) under the jurisdiction of Xining City, Qinghai Province into 5 sampling areas according to east, west, south, north, and center each year. One township (town, street) was selected from each area. Forty non boarding students aged 8 to 10 from each primary school (half male and half female, age balanced) and 20 pregnant women from each township (town, street) location were selected to collect edible salt samples at home and a random urine sample to measure salt iodine and urinary iodine level. B-ultrasound was used to measure thyroid volume in children and the goiter rate was calculated.Results:A total of 6 534 samples of household edible salt were collected from children and pregnant women, with an average salt iodine concentration of 25.58 mg/kg. The coverage rate of iodized salt was 97.50% (6 371/6 534), and the qualified iodized salt consumption rate was 89.46% (5 845/6 534). A total of 4 362 urine samples were collected from children, with a median urinary iodine level of 183.10 μg/L. The difference between different years was statistically significant ( H = 20.27, P < 0.001). A total of 2 169 urine samples were collected from pregnant women, with a median urinary iodine level of 168.90 μg/L. The difference between different years was statistically significant ( H = 107.09, P < 0.001). A total of 3 336 cases of thyroid gland examination were conducted in children, including 33 cases of thyroid enlargement, with a goiter rate of 0.99%. There was a statistically significant difference between different years (χ 2 = 15.00, P < 0.001). Conclusion:From 2019 to 2021, children aged 8 to 10 and pregnant women in Xining City are at an appropriate level of iodine, and the achievements in prevention and treatment of iodine deficiency disorders still need to be continuously consolidated.