Correlation of Folic Acid Metabolic Gene Polymorphisms, Homocysteine, Vitamin B12, and Red Blood Cell Folate with Adverse Pregnancy Outcomes.

Objective: This study aims to investigate the relationship between folic acid (FA) metabolic gene polymorphisms, homocysteine (Hcy), vitamin B12 (Vit B12), and red blood cell folate (RBCF) with adverse pregnancy. The findings of this study can help in the prevention and treatment of adverse pregnancy in the future. Methods: 118 pregnant women admitted to Qingdao Central Hospital between August 2020 and October 2022 were selected for retrospective analysis, including 62 cases of normal delivery (control group, CG) and 56 cases of adverse pregnancy (research group, RG). The single nucleotide polymorphisms of MTHFR C677T, MTHFR A1298C, and MTRR A66G gene loci were tested in both cohorts. Besides, differences in Hcy, Vit B12, and RBCF levels were observed, as well as Hcy, Vit B12, and RBCF alterations in different genotype carriers in the research group. Results: An elevated proportion of MTHFR 677TT-type gene and MTRR 66GG-type gene carriers and a lower proportion of MTRR 66GG-type gene carriers were found in the research group (χ2 = 4.458, 4.238, 4.206, P = .035, .040, .040). As indicated by the Logistic regression analysis, carriers of MTHFR 677TT and MTRR 66GG gene had an increased risk of adverse pregnancy outcomes (95%CI=2.881-5.942, 1.427-3.809, P < .001), while MTRR 66AG carriers had a decreased risk (95%CI=0.124-1.849, P < .001). Finally, Hcy levels of MTHFR 677TT and MTRR 66GG gene carriers increased, while Vit B12 and RBCF decreased; the opposite was true for MTRR 66AG gene carriers (P < .001). Conclusions: FA metabolic gene polymorphisms, Hcy, Vit B12, and RBCF are closely related to adverse pregnancy outcomes, which is of great significance for future clinical evaluation of adverse pregnancy.

The chromosome-scale reference genome of Rubus chingii Hu provides insight into the biosynthetic pathway of hydrolyzable tannins.

Rubus chingii Hu (Fu-Pen-Zi), a perennial woody plant in the Rosaceae family, is a characteristic traditional Chinese medicinal plant because of its unique pharmacological effects. There are abundant hydrolyzable tannin (HT) components in R. chingii that provide health benefits. Here, an R. chingii chromosome-scale genome and related functional analysis provide insights into the biosynthetic pathway of HTs. In total, sequence data of 231.21 Mb (155 scaffolds with an N50 of 8.2 Mb) were assembled into seven chromosomes with an average length of 31.4 Mb, and 33 130 protein-coding genes were predicted, 89.28% of which were functionally annotated. Evolutionary analysis showed that R. chingii was most closely related to Rubus occidentalis, from which it was predicted to have diverged 22.46 million years ago (Table S8). Comparative genomic analysis showed that there was a tandem gene cluster of UGT, carboxylesterase (CXE) and SCPL genes on chromosome 02 of R. chingii, including 11 CXE, eight UGT, and six SCPL genes, which may be critical for the synthesis of HTs. In vitro enzyme assays indicated that the proteins encoded by the CXE (LG02.4273) and UGT (LG02.4102) genes have tannin hydrolase and gallic acid glycosyltransferase functions, respectively. The genomic sequence of R. chingii will be a valuable resource for comparative genomic analysis within the Rosaceae family and will be useful for understanding the biosynthesis of HTs.

Colorimetric grading scale can promote the standardization of experiential and sensory evaluation in quality control of traditional Chinese medicines.

Experiential and sensory evaluation is an ancient method that remains important in the current quality control system of Traditional Chinese Medicines (TCMs). The process is rapid and convenient when evaluating the quality of crude materials in TCM markets. However, sensory evaluation has been met with skepticism because it is mainly based on experience and lacks a scientific basis. In this study, rhubarb was selected to demonstrate how color-based sensory evaluation could differentiate the quality of herbal medicines objectively. The colors of the rhubarb samples, expressed as RGB values, were obtained from different parts and forms of the plant, including the plant's surface, fracture surface color, and a powdered form with or without treatment with a color-developing reagent. We first divided the rhubarb samples into three grades based on the total content of five hydroxyanthraquinone derivatives, the major pharmacological components in rhubarb. Then, a three-layer back-propagation artificial neural network (BP-ANN), calibrated with selected training samples, was used to correlate the quality of the rhubarb with its color. The color of the rhubarb powder after coloration attained the highest accuracy (92.3%) in predicting the quality grade of the test samples with the established artificial neural networks. Finally, a standardized colorimetric grading scale was created based on the spatial distribution of the rhubarb samples in a two-dimensional chromaticity diagram according to the colors of the powdered rhubarb after color enhancement. By comparing the color between the scale and the tested samples, similar to performing a pH test with indicator paper, subjects without sensory evaluation experience could quickly determine the quality grade of rhubarb. This work illustrates the technical feasibility of the color-based grading of rhubarb quality and offers references for quantifying and standardizing the sensory evaluation of TCMs, foods and other products.

[Research on good usage practice for Chinese Materia Medica (I): chemical equivalence of different prepare procedures and optimal conditions for good clinical usage of rhubarb].

OBJECTIVE: To probe into the influences of different granule sizes and to prepare procedures on rational clinical usage of rhubarb based on chemical equivalence. METHOD: The effects of particle size, extract solvent, extract time and repeat times, and pre-extract or pro-extract of rhubarb on the extract amounts of the anthraquinones (AQs) were compared. RESULT: The different prepare procedures investigated in the paper revealed significant influence on the extract amounts of the AQs and those extracts were not chemical equivalent. Ethanol extracted more AQs than water did, when other conditions were same. When extracted with water, the rhubarb of piece size 0.8-1.2 cm could extract relatively high amount of AQs nearly equal to superfine grinded powders, and the former was cheap. The water extraction of AQs showed an increasing trend with the extraction time extended. And pro-extract manner with water could extract more AQs than pre-extract manner with a extraction time of 30, 60 min. The water extraction of AQs repeated two times exceeded half of the amount of totally six times. When extracted with ethanol, the rhubarb of fine powders could extract relatively high amount of AQs nearly equal to superfine grinded powders. And pre-extract manner with ethanol could extract more AQs than pro-extract manner. The ethanol extraction of AQs increased in 30 min and then increased slower. The ethanol extraction of AQs repeated two times exceeded 70% of the amount of totally six times. So, the optimal conditions for water extraction rhubarb were pro-extract, two times repeated and 30 min per time; and the optimal conditions for ethanol extract were pre-extract, two times repeated and 30 min per time. CONCLUSION: The different prepare procedures showed significant influence on the extraction of rhubarb AQs. There is great need to establish a good usage practice (GUP) for Chinese Materia Medica to maintain rational clinical usage.

[Novel method for dynamic monitoring and early-warning on wild resources of traditional Chinese medicines based on price ratio between drug and foodstuff].

OBJECTIVE: To probe into the new idea along with establishment of a novel method for dynamic monitoring and early-warning on the wild resources of traditional Chinese medicines (TCMs). METHOD: The alterations of wild traditional Chinese medicinal resources were assessed through the price ratio between drug and foodstuff (PRDF) indicating the balance between supply and demand of the specific TCMs, referred to the price ration between pork to foodstuff which is used in national monitoring to the balance between pork supply and demand. RESULT: Since the price of rice was tightly controlled by government, it was selected as a relatively stable reference to build the PRDF in order to take away the non-marketing influence to TCMs price such as CPI and inflation rate. The modified relative alteration trend of TCMs price had been researched through comparing different formulae to build PRDF, including absolute average month price of TCMs, month average price ratio of TCMs to foodstuff (rice) , month-on-month change of TCMs to rice, year-on-year change of TCMs to rice, and difference in value of period-on-period change (DVPPC). In the research, Cordyceps, Glycyrrhiza and totally five herbs were selected as model drugs and the price data were collected from 2002 to 2008. The results showed that DVPPC calculated of relative long time window was more sensitive and stable to reflect the relative alteration trend of TCMs price. For instance, the DVPPC of Ligustici showed continuously increase trend in recent years. This suggested appearance of unbalance between supply and demand of Ligustici, and forced policy intervention to maintain reasonable and continuable utilization of Ligustici resource. CONCLUSION: The proposed method and the formula of DVPPC revealed some useful guidance for dynamic monitoring the wild resources of TCMs.

Study on the cold and hot properties of medicinal herbs by thermotropism in mice behavior.

It is a common sense that chewing a mint leaf causes a cold feeling, while masticating a piece of ginger root is associated with a hot sensation. The Traditional Chinese Medicine has termed this phenomenon as cold and hot properties of herbs and applied them in treating certain human diseases successfully for thousands of years. Here, we have developed an Animal Thermotropism Behavior Surveillance System, and by using this device and other approaches, we not only verified the existence of, but also characterized and quantitated the cold and hot properties of medicinal herbs in animal behavioral experiments. The results suggested that the hot and cold properties of herbal drugs indeed correlated with the alteration of animal behavior in search for residence temperature.

Clinical study on the treatment of knee osteoarthritis of Shen-Sui insufficiency syndrome type by electroacupuncture.

OBJECTIVE: To study the clinical effificacy of electroacupuncture (EA) on treating knee osteoarthritis (KOA) of Shen ()-Sui () insuffificiency (SSI) syndrome type. METHODS: A total of 245 patients (279 knees) of KOA-SSI were randomly assigned to two groups by lottery: 141 knees in the treatment group and 138 knees in the control group. The treatment group was managed with EA at the dominant points of Neixiyan (Ex-LE4) and Waixiyan (Ex-LE5) as well as the conjugate points of Xuanzhong (GB39) and Taixi (KI3) for 30 min, once a day, with 15 days as one course; 2 courses were applied with a 5-day interval in between. The control group was treated with intra-articular injection of 2 mL hyaluronic acid into the affected joint every 7 days for 5 times in total. The clinical effects on the patients in different stages were observed, and their symptom scores of knee and contents of cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), prostaglandin E(2alpha) (PGE(2alpha)) and matrix metalloproteinases-3 (MMP-3), in the knee joint fluid were measured before and after treatment. RESULTS: The study was completed in 235 patients (263 knees); four patients (7 knees) in the treatment group and six patients (9 knees) in the control group dropped out. Comparison of therapeutic effects (excellent and effective rates) between the two groups showed insignificant differences (P>0.05). Symptom scores of knee and contents of cytokines in the knee flfluid after treatment were lowered signifificantly in the patients of stage I-III in both groups (P<0.05 or P<0.01). However, the lowering of the total symptom score of knee in the patients of stage III in the treatment group was more signifificant (P<0.05). CONCLUSIONS: EA could effectively alleviate the clinical symptoms in KOA patients of stage III, showing an effect superior to that of hyaluronic acid. EA also shows action in suppressing the secretion of IL-1, IL-6, TNF-alpha, PGE(2alpha) and MMP-3 in the knee flfluid.

[Rationality of commercial specification of rhubarb based on chemical analysis].

The differences of 34 rhubarb samples collected on the market and at producing area were investigated by chemical analysis on the contents of anthraquinones and chromatographic fingerprints, in order to assess the rationality of the commercial specification of rhubarb. The results indicated that the commercial specification of rhubarb was not correlated to the contents of anthraquinones as well as the price. The chromatographic fingerprints of rhubarb samples from different producing area were dissimilar, while the commercial specifications were difficult to be separated. Generally, the rhubarb samples produced in famous-region contained more anthraquinones. This demonstrated rationality on the traditional records of the famous-region of rhubarb from a chemical view. In this study, it was firstly reported that rhubarb could be categorized into two types, chrysophanol-type and rhein-type, based on the proportion of the two constituents in the total anthraquinones after acid hydrohysis. It was found that the rhubarb samples of rhein-type were mostly produced in famous-regions, such as Qinghai, Xizang, West Sichuan and Gansu. The literatures reported that rhein was superior to chrysophanol at many pharmacological effects and pharmacokinetic properties. Hence, we primarily considered that rhein-type rhubarb might be high-quality. These results were helpful to improve the commercial specification of rhubarb from a view of chemical information.

[Delphi-based study on scientificity of organoleptic evaluation for rhubarb commercial specification].

OBJECTIVE: In this study the reliability and scientificity of organoleptic evaluation for the commercial specification of Chinese medicinal materials (CMM) was investigated with Delphi method, using rhubarb as an example. The consensus panel of panelists being expert in organoleptic evaluation for CMM was cogitatively selected, with high average authority index of 0.81. The average individual repeatability of selected panelists was 85%, while the group repeatability was above 70%. It was found that the career time of the expert had significant influence on the evaluation result (P < 0.05), which suggested that practice makes repeatability. Some panelists of different career type and education background concluded different results, but no statistical significance was found. Furthermore, four predominant organoleptic parameters, shape, color, texture and section characteristics, were selected based on statistics and panelists' discussion to judge the commercial specification of rhubarb. In conclusion, we firstly demonstrated the organoleptic evaluation is an objective and repeatable approach to judge the commercial specification of CMM; and we recommend Delphi method to do validation and standardization for the conventional experiences of traditional Chinese medicine.

[Antimicrobial activity and chemical differences between the two chemotypes of rhubarbs].

Through our pre-investigation and literature analysis, it was found that rhubarb could be categorized into two types, chrysophanol-type and rhein-type, based on the proportion of the two constituents in the total content of anthraquinones after acid hydrolysis. In this paper, the antimicrobial activities of chrysophanol-type and rhein-type rhubarbs against Staphylococcus aureus were compared with microcalorimetric analysis, in order to illustrate the bioactive differentiability between the two chemotypes. For the aim to display the distinction of chrysophanol and rhein percentage in total anthraquinones, the sampling volume was regulated to make the total anthraquinones equivalent, thus, the antimicrobial difference was only attributed to the difference of chemotypes. The results indicated that the antimicrobial difference between the two chemotypes was confirmable labeled at the biothermokinetic parameters of S. aureus growth affected by the rhubarb samples. The growth rate constant (k1) of the first exponential phase for the growth of S. aureus affected by the rhein-type rhubarb was significantly lower than that of chrysophanol-type (P<0.01), which suggested stronger antimicrobial activity of rhein-type rhubarb than that of chrysophanol-type. However, the antimicrobial activities of rhein-type rhubarbs were not positively correlated to the contents of rhein. It suggested that the antimicrobial activity of rhubarb might be related to some unknown components which were of same accumulating pattern of rhein. The findings in present study provided some experimental evidence on categorizing rhubarb into two chemotypes through the difference of antimicrobial activity on S. aureus by microcalorimetric analysis and, further, offered references to revision of the commercial specification of rhubarb from chemical view.

[Differences between cold and hot natures of processed Radix ginseng rubra and Panax quinquefolius L. based upon mice temperature tropism].

OBJECTIVE: To establish an objective method to estimate the disparity between the cold and hot natures on the basis of an intrinsic correlation between temperature tropism of mice and the cold and hot natures of Chinese medicines. METHODS: Male KM mice were randomly divided into 7 groups of 6 each, namely the normal group (NM), the weak model group (WM), the strong model group (SM), the weak model plus Radix ginseng rubra group (WM + RG), the weak model plus Panax quinquefolius L. group (WM + PQ), the strong model plus Radix ginseng rubra group (SM + RG) and the strong model plus Panax quinquefolius L. group (SM +PQ). The specific herbal drugs were administered intragastricly. To induce the weak model, mice were fed with a limited supply of feed and forced to swim in cold water until almost drowning while the strong model induced by feeding a high-protein diet with an unlimited feed access. The doses of Radix ginseng rubra and Panax quinquefolius L. were 35 mg/g of body weight per day (counted by the quantity of crude material) and lasting for seven days. The NM and model groups without dosing were intragastricly administered with physiological saline of the same volume to the dosing groups. The percentage of the remaining time of mouse on a high temperature (40 degrees C) pad to the total monitoring time was recorded by a self-designed intelligent animal behavior monitoring system. Meanwhile, the drinking volume of mice in each group was measured. Immediately after experiment, the activities of Na(+)K(+)-ATPase and superoxide dismutase (SOD) in liver tissue were measured by assay kits of phosphorus and xanthine oxidase methods respectively. RESULTS: The features of deficient and cold symptom, such as fatigue, stagnant weight growth, decreased water intake, cold limbs and tail etc, were observed in WM group. And the features of heat symptom, such as increased weight and water intake, hyperactivity etc, were observed in SM group. The percentage of time that the mouse remained on 40 degrees C pad of the WM group within the seven days experiment was significantly higher than that of the normal group (70.6% +/- 21.3% vs 52.1% +/- 6.5%, P < 0.05). While the value of SM group (45.7% +/- 4.6% ) was significantly lower than that of the normal group (P < 0.05); the value of WM + RG group and WM + PQ group were 65.6% +/- 7.8% and 75.3% +/- 13.0% respectively (both P < 0.05 compared with WM group); the values of SM + RG group and SM + PQ group were 36.1% +/- 15.5% and 55.5% +/- 7.7% respectively (both P < 0.05 compared with SM group). The activities of Na(+)K(+)-ATPase and SOD of WM mice treated with either Radix ginseng rubra or Panax quinquefolius L. were found to have a significant up-regulation (P < 0.05) as compared with untreated WM mice. But only the Panax quinquefolius L. showed an up-regulating effect upon Na(+)K(+) ATPase and SOD in SM mice. CONCLUSIONS: The external cold and hot natures of Radix ginseng rubra and Panax quinquefolius L. can be represented in an ethological way by the changes of animal's temperature tropism. And such a tropism may be internally regulated by body's energy metabolism.

Investigation of the differences between the "Cold" and "Hot" nature of Coptis chinensis Franch and its processed materials based on animal's temperature tropism.

The description and differentiation of the so-called "Cold" and "Hot" natures, the primary "Drug Naure" of Chinese medicine, is the focus of theoretical research. In this study, the divergency between the "Cold" and the "Hot" natures was investigated through examining the temperature tropism of mice affected by Coptis chinensis Franch and its processed materials by using a cold/hot plate differentiating technology. After exposure to C. chinensis Franch, the macroscopic behavioral index of the remaining rate (RR) on a warm pad (40 degrees C) significantly increased (P<0.05), suggesting the enhancement of Hot tropism. The internal indexes of adenosine triphosphatase (ATPase) activity and oxygen consuming volume decreased significantly (P<0.05), suggesting the decapability of energy metabolism. This external behavior of Hot tropism might reflect the internal Cold nature of C. chinensis Franch. However, the processed materials of C. chinensis Franch exhibited a different Cold nature in temperature tropism compared with crude C. chinensis Franch (CC): the Cold nature of bile-processed C. chinensis Franch (BC) enhanced while the ginger-processed C. chinensis Franch (GC) changed inversely. The changing sequence was consistent with the theoretical prognostication. It is indicated that the external Cold & Hot natures of Chinese medicine may possibly reflect in an ethological way for the changes of animal's temperature tropism which might be internally regulated by the body's energy metabolism.

[COLD and HOT nature of Coptis & Evodia and their prescriptions investigated with diet restriction/cold-water swimming mice models].

To establish a new method to evaluate the COLD and HOT nature of Coptis & Evodia and their prescriptions Zuojinwan and Fanzuojinwan. Physical models of mice were established by diet restriction with cold-water swimming (weak model, WM) and fed with high protein animal feeds (strong model, SM). An instrument with cold and hot pads was used to investigate the variation of temperature tropism among SM and WM groups of mice affected by drugs. Meanwhile, the oxygen consumption and activity of adenosine triphosphatase (ATPase) were detected, in order to investigate the mechanism of energy metabolism which might be affected by these drugs. The results showed that the drug effects gradually changed in an order of "Coptis-->Zuojinwan--> Fanzuojinwan-->Evodia". In detail, Coptis increased the remaining rate (RR) of mice on hot pad, decreased oxygen consumption and ATPase activity (n=6, P < 0.01 or P < 0.05), while Evodia performed inversely; which indicated the COLD nature of Coptis and HOT nature of Evodia, and confirmed with their traditional definition in medicinal works. In conclusion, the methods applied in this work, can objectively and directly express the nature disparity between the two herbs and predict the tendency of changes of the nature of their combination, which brings a new approach in investigation of the nature theory of traditional Chinese medicine.