ObjectiveTo explore the impacts of material type and loading volume of rigid containers on the hydrogen peroxide low temperature plasma sterilization of thoracoscopic instruments, to identify the best rigid containers and loading volume of thoracoscopic instruments. MethodsThoracoscopic instruments sterilized by STERRAD^® 100NX hydrogen peroxide low temperature plasma in Shanghai Pulmonary Hospital affiliated to Tongji University from August to September 2024 were selected as the research items. According to the material of rigid containers, the instruments were divided into polyethylene case group (A), stainless steel case group (B) and silicone resin case group (C). In terms of the loading volume, the rigid containers were divided into (loading capacity <80%) groups of 8, 10 and 12 instruments. The results of physical monitoring, the first type of chemical indicator card monitoring, and the five types of card luminal chemical process challenge device (PCD) monitoring of the 9 groups of A8, A10, A12, B8, B10, B12, C8, C10 and C12 were compared and evaluated. ResultsCompared to A8, A10 A12, C8, C10 or C12 groups, the thoracoscope instruments in the stainless steel containers in B8, B10 or B12 group had higher hydrogen peroxide concentrations and shorter elapsed time in the pressure check phases 1 and phases 2, with the differences statistically significant (P<0.05), followed by the silicone resin case group and the polyethylene case group. The nine groups of physical parameter monitoring, the first type of chemical indicator monitoring, and the five types of chemical PCD monitoring for lumen sterilization achieved 100% qualification rates, and there were no significant differences in the qualified rates of sterilization among the 9 groups (P>0.05). ConclusionWhen using hydrogen peroxide low temperature plasma to sterilize thoracoscopic instruments, it is recommended to use stainless steel or silicone resin rigid containers with a controlled loading capacity (≤12) to ensure optimal sterilization quality.

Background::Radiation (IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment (TME). Wee1, a cell cycle regulator, can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1 (CDK1). Meanwhile, programed death-1/programed death ligand-1 (PD-1/PDL-1) blockade is closely related to TME. This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody (anti-PD-1 Ab) on radiosensitization of hepatoma.Methods::The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay on human and mouse hepatoma cells HepG2, Hepa1-6, and H22. The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro. A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice, which were divided into control group, IR group, AZD1775 group, IR + AZD1775 group, IR + anti-PD-1 Ab group, and the IR + AZD1775 + anti-PD-1 Ab group. Cytotoxic CD8 + T cells in TME were analyzed by flow cytometry. Results::Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro. AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage. AZD1775 treatment also reduced the proportion of PD-1 +/CD8 + T cells in the spleen of hepatoma subcutaneous xenograft mice. Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferon γ (IFNγ) + or Ki67 + CD8 T cells and decreasing the levels of CD8 + Tregs cells in the tumor and spleen of the hepatoma mice model, indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγ expression, enhancing CD8 + T cells proliferation, and weakening CD8 + T cells depletion. Conclusions::This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improving cytotoxic CD8 + T cells in TME.

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGAD^{S24R/D88R/Y309K} was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and Lpgad^{S24R/D88R/Y309K} genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD₆₀₀) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.
Glutamate Decarboxylase/genetics* ; Lactobacillus plantarum/genetics* ; Catalysis ; gamma-Aminobutyric Acid ; Hydrogen-Ion Concentration ; Glutamic Acid

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

OBJECTIVE：To investigate the effects of pharmaceutical management intervention on the use of TCM injection in primary medical institutions. METHODS ：Twenty grass-roots health centers in Yichun city ，Jiangxi province were randomly selected. Relying on Department of Pharmacy and Health Services of the Municipal Health Committee and the Quality Control Center for Pharmacy Affairs of the City ，Yichun People ’s Hospital took the lead in establishing the prescription evaluation criteria for TCM injection ，training pharmaceutical service skills ，conducting special reviews on prescriptions/medical orders ，improving control measures ，obtaining relevant data of drug use in primary health centers by issuing questionnaires on the use of TCM injection and randomly selecting prescriptions/medical orders for comments. The relevant data without drug administration （before intervention）from Jan. to Jun. 2018 were taken as the non-intervention group ，and the relevant data with drug administration （after intervention）from Jan. to Jun. 2019 were taken as the intervention group. The use of TCM medicine injection ，DDDs，DDC and sequencing ratio were compared before and after intervention. At the same time ，the prescriptions/medical orders of TCM injections before（1 222 copies）and after （1005 copies）intervention were randomly selected for comment and the reasonable situations of the prescriptions/medical orders were compared before and after the intervention. RESULTS ：After intervention ，total amount ， consumption sum and DDDs of top 10 TCM injections in the list of amount were significantly decreased. The reasonable rate of prescriptions and medical orders was 48.04％ before intervention ， but increased to 70.65％ after intervention. Besides ， unreasonable situations before and after intervention all manifested as inappropriate indications. After the intervention ，the unreasonable rates of Xuesaitong powder for injection （lyophilized），Xueshuantong injection ，Xingnaojing injection （two specifications），Shengmai injection ，Xueshuantong for injection （lyophilized）and Qingkailing injection were significantly lower than before intevention （P＜0.05）. CONCLUSIONS ：Pharmaceutical administration intervention can reduce the total amount and the consumption sun of TCM injection in primary medical institutions ，and promote rational drug use in the clinic.

Objective:To understand the basic nutrition situation of primary and secondary school students in the urban area of Yichun City， and to provide corresponding strategies and measures for the improvement of the nutrition of children and adolescents. Methods:A stratified sampling method was used to select the urban primary and secondary school students in 10 counties （cities， districts） in Yichun City from 2019 to 2020 for physical examinations to assess their nutritional status. Results:Among 8 921 primary and middle school students surveyed， 62（0.69%） were stunted in growth， 735（8.24%） were emaciated， 1 130（12.67%） were overweight， and 662（7.42%） were obese. The weight loss rate of primary school students was the highest （10.54%， 360/3 414）， followed by junior middle school students （7.07%， 214/3 026）， and senior high school students （6.49%， 161/2 481）. The difference was statistically significant （χ²=39.51， P<0.05）. The proportion of unbalanced nutritional status of boys was 35.66% （1 578/4 425）， which was higher than that of girls （22.49%， 1 011/4 496）， and the difference was statistically significant （χ²=187.91， P<0.01）. Conclusion:Both malnutrition and overweight/obesity exist in primary and middle school students in Yichun City. We should pay special attention to boys and lower grade students， and intervene different nutritional problems.

Scutellariae Radix is a kind of traditional Chinese medicine, which has the functions of heat-clearing and damp-drying, purging fire and detoxifying, hemostasis and miscarriage prevention. Modern pharmacological studies show that Scutellariae Radix has various effects, such as anti-oxidation, anti-inflammatory, liver protection and antiviral microorganisms. By searching the documents in the past ten years, the author has found that Scutellariae Radix and its active components play an important role in protecting the liver. It can prevent and cure liver injuries caused by different reasons, including acute or chronic hepatitis, liver fibrosis and liver cancer. Among all kinds(chemical, immunological, alcoholic, nonalcoholic, viral, ischemia-reperfusion, etc.) of acute or chronic hepatitis, most studies are on CCl_4 induced chemical liver injury. Scutellariae Radix and its active components can significantly reduce the serum transaminase level in hepatitis animals, and reduce the degree of liver pathological damage. The mechanisms include antioxidant stress, anti-inflammatory, anti-apoptosis, inhibition of immunity, anti-virus and regulation of lipid metabolism, etc. Scutellariae Radix and its active components can significantly inhibit the activation of hepatic stellate cells and reduce extracellular matrix, and its anti-fibrosis mechanism involves antioxidation, anti-inflammatory, inducing apoptosis of hepatic stellate cells and so on. Whether in vivo or in vitro, Scutellariae Radix and its active components show a good anti-hepatocarcinoma effect, especially on hepatocarcinoma. Its prevention and treatment mechanisms for hepatocarcinoma mainly include blocking cancer cell cycle, inhibiting cancer cell metastasis, promoting cancer cell apoptosis and inducing autophagy. It can be seen that Scutellariae Radix has multiple functions and mechanisms in liver protection, and has a great development potential. Therefore, this paper reviews the prevention and treatment effects and mechanism of Scutellariae Radix and its components on different liver diseases, in order to provide reference for in-depth study, development and clinical application in the prevention and treatment of liver disease.
Animals ; Anti-Inflammatory Agents ; Drugs, Chinese Herbal ; Flavonoids ; Medicine, Chinese Traditional ; Scutellaria baicalensis

To detect the concentration of triptolide in skin and joint after percutaneous administration,an HPLC-MS/MS method and skin and joint micro-dialysis( MD) method of triptolide were established in this study. The separation was achieved on triple quadrupole( AB QTRAP4500) and phenomenex-C18( 4. 6 mm×150 mm,5 μm,luna) column with acetonitrile-water with 0. 1% formic acid( 65 ∶35) as the mobile phase at a flow rate of 0. 7 m L·min-1. An electrospray ionization( ESI) source was applied and operated in the positive multiple reaction monitoring( MRM) mode. The fragment ion for triptolide was m/z 361. 1→145. 0. The effects of different perfusion [Ringer's,PBS( p H 7. 4),30% ethanol saline]drug concentrations and flow rates on the recovery rate,as well as the relationship between the recovery rate and the loss rate were determined by incremental( dialysis) and reduction( retrodialysis) methods.The reduction method was applied in the in vivo study to investigate and determine the stability of the probe recovery rate in 10 h. The results of HPLC-MS/MS detection method conformed to the requirements of biological samples. The perfusion fluid was 30% ethanol saline. The recovery rate of skin and joint probes in vitro of triptolide increased within the flow rate of 0. 5-2. 5 μL·min-1. In order to increase the timeliness of data and the accuracy,the flow rate was determined to be 1 μL·min-1,and the sample interval was determined to be 0. 5 h. The recovery rate of triptolide in skin and joint probes in vitro and the loss rate were stable and equal despite of change of triptolide concentration within 10-200 μg·L-1. This indicated that the effect of drug concentration on the MD probe recovery rate was small,and the recovery rate could be replaced by the loss rate. The loss rate in vivo using MD method was measured at 10 h,indicating that the transfer rate of triptolide was stable within 10 h. The established method of triptolide in MD and HPLC-MS/MS can be applied to investigate the kinetic in skin and joint after percutaneous administration of triptolide.
Chromatography, High Pressure Liquid ; Diterpenes ; pharmacokinetics ; Epoxy Compounds ; pharmacokinetics ; Humans ; Joints ; metabolism ; Phenanthrenes ; pharmacokinetics ; Reproducibility of Results ; Skin ; metabolism ; Tandem Mass Spectrometry

Objective: This study is designed to investigate the effect and mechanism of Pingkui enema in treatment of ulcerative colitis. Methods: Sixty SD rats at SPF level were randomly divided into 6 groups which were normal group, model group, and the low, medium and high dose groups of Chinese medicine Pingkui enema, and sulfasalazine (SASP) enema group. The normal group and model group were given an equal volume of saline enema after modeling. After continuous administration for 10 days, the histopathological effects were evaluated after extraction, and the serum interleukin-8 (IL-8), interleukin-13 (IL-13), tumor necrosis factor (TNF-α), rat intestinal mucosal bifidobacteria adhesion and adhesin receptor were measured by enzyme-linked immunosorbent assay (ELISA). The real-time fluorescence quantitative PCR (RT-PCR) were used to detect the intestinal bifidobacterial content. Results: Compared with the model group, the content of IL-8 (153.50 ± 8.59 pg/ml, 150.84 ± 5.38 pg/ml vs. 167.13 ± 13.66 pg/ml), TNF-α (330.08 ± 10.02 pg/ml, 287.27 ± 13.89 pg/ml vs. 376.50 ± 19.06 pg/ml) in medium, high dose group of Pingkui enema significantly decreased (P<0.05), while the IL-13 (37.04 ± 3.62 pg/ml, 40.64 ± 5.98 pg/ml vs. 31.57 ± 2.95 pg/ml) significantly increased (P<0.05). The bifidobacteria adhesin (124.84 ± 9.74 ng/L, 149.93 ± 9.57 ng/L, 176.74 ± 11.01 ng/L vs. 101.11 ± 17.18 ng/L) and adhesin receptor (78.44 ± 14.03 ng/L, 99.50 ± 3.63 ng/L, 107.36 ± 5.05 ng/L vs. 60.96 ± 13.89 ng/L) in low-, medium-, high dose group of Pingkui enema significantly increased (P<0.05). The content of bifidobacteria (1 331.4 ± 242.2 copies/μl vs. 490.5 ± 106.0 copies/μl) in the middle dose Pingkui enema group significantly increased (P<0.05). Conclusions The Pingkui enema can promote the colonization of intestinal epithelial cells by promoting bifidobacteria, increase bifidobacterium adhesin and adhesin receptors, up-regulate the expression of IL-13, and down-regulate the expression of IL-8, TNF-α.

OBJECTIVETo compare the effects between acupoint embedding and oral medication for female functional constipation.

METHODSFifty-six female patients were randomized into an embedding group and a medication group (1:1).There were 22 cases in the embedding group and 26 cases in the medication group with total 8 patients dropped out. The main embedding acupoints were Zhongwan (CV 12), bilateral Tianshu (ST 25) and Guanyuan (CV 4), combined with Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Zhigou (TE 6) and Shangjuxu (ST 37). It was given once a week. Oral lactulose was applied in the medication group, 3 times a day. The treatment cycle was 8 weeks. The primary outcome was the percentage of patients whose weekly average number of complete spontaneous bowel movement (CSBM) was increased ≥ 2 compared with baseline during the last 6 weeks in the treatment period. The secondary efficacy indices were compared before and after treatment as well as at follow-up at the 12th week, including the weekly average number and increased number of CSBM, the quality of life of patients with constipation (PAC-QOL), the Bristol stool character score, and the assessment of difficulty in defecation.

RESULTSThe percentage of patients whose weekly average number of CSBMs were increased ≥ 2 in the embedding group significantly increased compared with that in the medication group (<0.05). The weekly average number of CSBM in the two groups increased after treatment compared with those before treatment (both<0.01); the score of Bristol stool character improved (both<0.01); the scores of PAC-QOL and difficulty in defecation decreased (all<0.01). The increasing time of weekly average CSBM in the embedding group was higher than that in the medication group (<0.01); the score of PAC-QOL in the embedding group after treatment was better than that in the medication group (<0.01); the Bristol stool character and difficulty degree in the embedding group after treatment were superior to those in the medication group (both<0.05).

CONCLUSIONSThe acupoint embedding significantly improve the CSMB, Bristol stool character, the difficulty of defecation, and the quality of life for female patients with functional constipation.