Objective To explore the role of neogambogic acid in the characteristics of colorectal cancer stem cells (CRC-CSCs) through the Wnt/β-catenin signaling pathway. Methods The colorectal cells SW480 and HCT166 were divided into control group and neogambogic acid groups (1.5, 3, 6, and 12 μmol/L). The viability of CRC-CSCs was determined by MTT method, and spheroid and clone formation assays were used to assess the capacity of spheroid formation and self-renewal ability of the cells. The effects of neogambogic acid on the apoptosis and cell cycle of CRC-CSCs were evaluated by flow cytometry assays. Real-time quantitative PCR was used to detect the mRNA expression levels of relative markers (CD133, CD44, ALDH1, Oct4, and Nanog) of CRC-CSCs, and the protein expression levels of the self-renewal marker (PCNA), apoptosis markers (cleaved caspase-3 and cleaved caspase-9), and Wnt/β-catenin signaling pathway markers (p-GSK3β, GSK3β, β-catenin, and Wnt) were analyzed using Western blot. Results Compared with the control group, after neogambogic acid treatment, the viability of SW480 and HCT116 cells decreased (P<0.05), the spheroid forming ability and the clone numbers of CRC-CSCs decreased (P<0.001, P<0.01) but the cell apoptosis rate increased (P<0.01), and cell cycle was arrested in G₀/G₁ phase. Moreover, neogambogic acid downregulated the mRNA and protein expression of relative markers of CRC-CSCs (CD133, CD44, ALDH1, Oct4, and Nanog), PCNA, p-GSK3β, β-catenin, and Wnt (P<0.05) and upregulated the expression of cleaved caspase-3, cleaved caspase-9, and GSK3β (P<0.01). Conclusion Neogambogic can inhibit the stem cell properties of colorectal cells via inhibition of Wnt/β-catenin signaling pathway. As a result, neogambogic acid may be an attractive agent against colorectal cancer.

By systematically combing ancient and modern literature， this paper examined Tribuli Fructus and Astragali Complanati Semen（ACS） used in the famous classical formulas from the aspects of name， origin， production area， harvesting and processing， clinical efficacy， so as to provide a basis for the development of famous classical formulas containing such medicinal materials. The results showed that the names of Tribuli Fructus in the past dynasties were mostly derived from its morphology， and there were nicknames such as Baijili， Cijili and Dujili. The name of ACS in the past dynasties were mostly originated from its production areas， and there were nicknames such as Baijili， Shayuan Jili and Tongjili. Because both of them had the name of Baijili， confusion began to appear in the Song dynasty. In ancient and modern times， the main origin of Tribuli Fructus were Tribulus terrestris， and ancient literature recorded the genuine producing areas of Tribuli Fructus was Dali in Shaanxi and Tianshui in Gansu， but today it is mainly cultivated in Anhui and Shandong. The fruit is the medicinal part， harvested in autumn throughout history. There is no description of the quality of Tribuli Fructus in ancient times， and the plump， firm texture， grayish-white color is the best in modern times. Traditional processing methods for Tribuli Fructus included stir-frying and wine processing， while modern commonly used is purified， fried and salt-processed. The ancient records of Tribuli Fructus were spicy， bitter， and warm in nature， with modern research adding that it is slightly toxic. The main effects of ancient and modern times include treating wind disorders， improving vision， promoting muscle growth， and treating vitiligo. The mainstream base of ACS used throughout history is Astragalus complanatus. Ancient texts indicated ACS primarily originated from Shaanxi province. Today， the finest varieties come from Tongguan and Dali in Shaanxi. The medicinal part is the seed， traditionally harvested in autumn. Modern harvesting occurs in late autumn or early winter， followed by sun-drying. Ancient texts valued seeds with a fragrant aroma as superior， while modern standards prioritize plump， uniform and free of impurities. Traditional processing methods for ACS included frying until blackened and wine-frying， while modern practice commonly employs purification methods. In terms of medicinal properties， the ancient and modern records are sweet and warm in nature. Due to originally classified under Tribuli Fructus， its effects were thus regarded as equivalent to those of Tribuli Fructus， serving as the medicine for treating wind disorders， additional functions included tonifying the kidneys and treating vitiligo. The present record of its efficacy is to tonify the kidney and promote Yang， solidify sperm and reduce urine， nourish the liver and brighten the eye， etc. Based on the textual research results， it is suggested that when developing the famous classical formulas of Tribuli Fructus medicinal materials， we should pay attention to the specific reference object of Baijili， T. terrestris and A. complanatus should be identified and selected， and the processing method should be in accordance with the requirements of the formulas.

The clinical data of a case of compound oxidative phosphorylation deficiency type 10 (COXPD10) caused by a new site mutation of MTO1 gene in the Department of Pediatrics, Affiliated Hospital of Southwest Medical University on December 29, 2020 were retrospectively analyzed.The patient was a 2 months and 19 days old boy of Han nationality.The main clinical manifestations were shortness of breath, hyperlactic acidemia, hyperammonemia and brain damage.Cardiac hypertrophy was not obvious.Heterozygous mutations at c. 344delA and c. 1055C>T sites in the MTO1 gene have not been reported in domestic and foreign literature.COXPD10 caused by MTO1 gene mutations may result in diversified clinical manifestations due to inconsistent mutation sites.For hyperlactic acidemia with unknown predisposing factors, early genetic examination should be conducted to confirm the possibility of COXPD10.

Objective: To investigate the feasibility of long noncoding RNA (lncRNA)_AK096792 as a clinical predictor of bronchopulmonary dysplasia (BPD) in preterm infants. Methods: All the cord blood(2-5 mL) of very low birth weight (VLBW) preterm infants born in Huai′an First Hospital Affiliated to Nanjing Medical University were collected from December 1, 2015 to December 1, 2017.Moreover, the peripheral blood(2 mL) of those VLBW infants diagnosed with BPD was also collected.A total of 36 infants with BPD were collected.Another 36 cases of premature children with VLBW were chosen as control group according to random number table.The relative content of lncRNA_AK096792 in cord blood and peripheral blood was detected by using real-time quantitative PCR (qPCR). Additionally, the correlation of lncRNA_AK096792 levels between cord and peripheral blood of BPD infants was analyzed.The sensitivity and specificity of lncRNA_AK096792 for BPD were analyzed by using receiver operating curve test. Results: (1)LncRNA_AK096792 was a common, evolutionarily conserved, non-coding RNA present in both mouse and human.(2) The expression level of lncRNA_AK096792 in peripheral blood was significantly higher than that in cord blood in BPD group[(463.3±352.0)% vs.(50.0±37.5)%], and the difference was significant(P<0.001), and they were highly correlated (r=0.825, P<0.001). (3) The level of lncRNA_AK096792 in cord blood in BPD group was signi-ficantly higher than that in non-BPD group [(484.3±280.5)% vs.(101.2±28.6)%], and the difference was significant(P<0.001). (4)When lncRNA_AK096792 served as a clinical predictor for BPD, the specificity was 83.3%, the sensitivity was 75.6%, and an area under the receiver operating characteristic curve was 0.88(P<0.001). Conclusions LncRNA_AK096792 is highly correlated with the development of BPD.The level of lncRNA_AK096792 in umbilical cord blood of premature infants can be used as an early predictive marker for BPD, it calls for further study.

Objective To evaluate the efficacy of epidural anesthesia combined with different doses ropivacaine and sufentanil for stepwise labor analgesia in latent phase.Methods Two hundred and ten ASA Ⅰ or Ⅱ primiparas with a singleton and vertex presentation at full term in our hospital from February 201 5 to April 201 5 were randomized into seven groups (n =30 each):0.125% ropiva-caine with 0.5 μg/ml sufentanil (group 1);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm) (group 2);0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group3);0.1 5% ropivacaine with 0.5μg/ml sufentanil (group 4);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥ 3 cm)(group 5 );0.1%ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group 6);0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm) (group 7).The intensity of pain was assessed by visual analog scale (VAS).Meanwhile,1abor process,postpartum hemorrhage,Bromage score,postpartum adverse reactions and Apgar score of the neonates were also observed.Results No significant difference was found in VAS score after epi-
dural block between groups at each time.The latent period of group 2 and 3 were shorter than that in group 1 (P <0.05)and that of group 5 and 6 were shorter than that in group 4 (P <0.05);the ac-tive phase of group 4 were longer than that in group 1 (P <0.05 ).The postpartum hemorrhage of group 2 and 3 were less than that in group 1 (P <0.05),the postpartum hemorrhage of group 5,6 and 7 were more than that in group 2 (P <0.05)and group 3 (P <0.05).The motor nerve block of group 2 and 3 were slightly less than that in group 1 (P <0.05)and the motor nerve block of group 5,6 and 7 were slightly less than that in group 4 (P <0.05).There was no difference of the postpar-tum adverse reactions of maternal and Apgar score in the neonates.Conclusion The dosage of 0.075% or 0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm),while producing the exact analgesic effect,hardly interferes with the 1abor process,the amount of postpartum hemorrhage and the lower limb activity,thus they have no significant effect on the safety of the maternal and the infant.

Objective To study the effect of ursolic acid on human laryngeal carcinoma Hep‐2 cells invasion and to explore its mechanism .Methods Methabenzthiazuron(M TT ) was used to observe the proliferation of Hep‐2 cells treated with various con‐centrations of ursolic acid at different time .Boyden chamber was used to compare ursolic acid effect on cell invasion .Western blot was used to the measure the NF‐κB protein expression .Gelatin zymography was used to the activity of MMP‐9 and MMP‐2 .Results Ursolic acid inhibited human laryngeal carcinoma Hep‐2 cells growth ,its effect showed dose dependent and time dependent (P <0 .01) .Boyden chamber results revealed that after the ursolic acid treatment Hep‐2 cells ,invasion ability showed a concentration de‐pendent decreased (P< 0 .01) .Western blot results revealed that ursolic acid showed a concentration dependent decreased on human laryngeal Hep‐2 cells NF‐κB protein expression(P< 0 .01) .Gelatinases spectrum revealed that ursolic acid a concentration depend ‐ent decreased on human laryngeal Hep‐2 cells MMP‐9 and MMP‐2 cells activity (P< 0 .01) .Conclusion Ursolic acid inhibit human laryngeal cancer Hep‐2 cell proliferation and invasion ,its mechanism is related to down‐regulate MMP‐9 and MMP‐2 activity and the NF‐κB protein expression .