Identification of Bupleurum Seeds by ITS2 Genetic Barcoding and Seed Morphology / 中国实验方剂学杂志

Objective:To accurately identify Bupleurum seeds by traditional morphological identification method combined with DNA barcoding technique. Method:A total of 41 seed samples on the market were collected and 75 ribosomal DNA internal transcribed spacer 2 (ITS2) sequences of 15 varieties were downloaded from the GenBank database as experimental materials. The seeds were measured and observed by stereomicroscope and vernier caliper, and their 1 000-grain weights were calculated. Genomic DNA was extracted from the seeds and used as a template, and ITS2 sequences were amplified using polymerase chain reaction (PCR) and bidirectional sequencing. Species identification was conducted based on BLAST method, neighbor-joining (NJ) phylogenetic tree method, Kimura two-parameter model (K2P) genetic distance method, and secondary structure of ITS2 sequence. Result:There were slight differences in the length, width, cross-section, and 1 000-grain weight among Bupleurum seeds from different origins. The ITS2 sequences of B. chinense seeds had 2 intraspecific variable sites and 3 haplotypes, the maximum intraspecific genetic distance (0.009) was far smaller than the minimum interspecific genetic distance (0.032). B. chinense and B. scorzonerifolium in the NJ phylogenetic tree were clustered into independent branches with good monophyletic property. The secondary structure of ITS2 sequences could make up for the shortcomings of NJ tree in identifying variants. The collected 41 seeds included 30 B. chinense seeds, 3 B. scorzonerifolium seeds, 5 B. falcatum seeds, 2 B. marginatum var. stenophyllum seeds, and 1 B. smithii var. parvifolium seeds. Conclusion:The B. chinense seeds on the market have problems of diverse sources and chaotic origins. Based on the combination of ITS2 gentic barcoding and seed morphological identification, the Bupleurum seeds can be accurately identified, which provides scientific bases for establishing the quality standard of Bupleurum seeds, standardizing the cultivation of B. chinense, and solving the quality problems of B. chinense from the source, and provides a reference for the accurate identification of other medicinal plant seeds or seed medicinal materials.

Clinical Efficacy of Modified Zhibo Dihuangwan in Treatment of Purpura Nephritis with Deficiency of Liver and Kidney Yin in Children / 中国实验方剂学杂志

Objective:To observe the clinical efficacy of modified Zhibo Dihuangwan on henoch-schonlein purpura nephritis with deficiency of liver and kidney yin in children (HSPN) and its effect on immune inflammatory response and hypercoagulable state. Method:Totally 120 patients were randomly divided into observation group (60 cases) and control group (60 cases) by random number table. Patients in two group was orally given prednisolone acetate tablets, 1.5-2 mg·kg-1·d-1, 2 times. Four weeks later, the drug was taken orally every other day, and the dosage decreased gradually after 4 weeks. Besides, patients in control group was intravenously dripped with cyclophosphamide, 8-12 mg·kg-1·d-1, for 2 days, and stopped for 2 weeks before another treatment course. The treatment lasted for 6 months. In the control group,Dabuyin Wan was taken orally,3 g/time,3 times/d.Patients in observation group was also added with modified Zhibo Dihuang Wan, 1 doe/day. The treatment lasted for 6 months. Urine routine was tested once a month, and disappearance time and rate of hematuria and albuminuria were recorded. The 24 h urine protein quantification, levels of microalbuminuria (mAlb) and urinary β2-microglobulin (β2-MG) were assessed before and after treatment. Furthermore, deficiency of liver and kidney Yin was scored, and levels of T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+), fibrinogen (FIB), D-dimer (D-D), fibrin degradation products (FDP), interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-10 (IL-10) were detected. Result:The clinical efficacy in observation group was superior to that in control group (Z=2.078，P<0.05). Disappearance times of hematuria and albuminuria of children in observation group were shorter than those in control group (P<0.01). The disappearance rate of proteinuria in observation group was 90.48%（38/42）, which was higher than 69.77%（30/43） in control group (χ2=5.694，P<0.05). The 24 h urinary protein quantity, mAlb and levels of β2-MG, FIB, D-D and FDP in observation group were lower than those in control group (P<0.01). The levels of CD3+, CD4+, IL-2 and IFN-γ and the ratio of CD4+/CD8+ in observation group were higher than those in control group (P<0.05), while the CD8+, IL-4 and IL-10 were lower than those in control group (P<0.05). The efficacy in observation group was better than that in control group (Z=2.106，P<0.05). Conclusion:In addition to conventional western medicine therapy, modified Zhibo Dihuang Wan have an effect on HSPN with deficiency of liver and kidney Yin in children by promoting the disappearance of albuminuria and hematuria, shortening the course of disease, improving T lymphocyte subpopulation, reducing inflammatory reaction and correcting hypercoagulable state of blood, with better clinical efficacy and syndrome effect of traditional Chinese medicine.

Advances in Bioculture Technology of Bupleuri Radix and Biosynthesis Pathways of Saikosaponins / 中国实验方剂学杂志

Synthetic biology is an emerging discipline that analyzes the biosynthesis pathways of active constituents in traditional Chinese medicine and explores genes involved in biosynthesis. Bupleuri Radix is one of the most commonly used Chinese medicinal materials with remarkable medicinal value, its index component is saikosaponins, which has significant anti-inflammatory, anti-viral and anti-tumor activities. However, the current wild resources of Bupleuri Radix have been destroyed, and there were some problems in the process of artificial cultivation. The application of biological culture technology and synthetic biology can expand the sources of saikosaponins and protect resources of Bupleuri Radix. The culture conditions of different plants can be followed without a fixed pattern, and the biosynthetic pathways of different medicinal active ingredients are also inconsistent. At present, there is no review report on the culture technology of Bupleuri Radix and the research on the biosynthesis pathway of saikosaponins. This paper introduces the research progress of biological culture techniques, such as callus culture, adventitious root culture, hairy root culture and suspension cell culture used in synthetic biology, and the biosynthesis pathway of saikosaponins and its key enzyme functional genes. It is suggested to optimize the biological culture technology of Bupleuri Radix by referring to the tissue culture technology of other traditional root medicinal materials, so as to provide a reference for the in-depth study on the biosynthesis pathway and metabolic regulation of saikosaponins.

Clinical efficacy of two different tricuspid annuloplasty techniques in left cardiac valve surgery

Objective:To investigate the mid-and long-term clinical effects of tricuspid valvuloplasty with the implantation of an artificial plastic ring.Methods:Data of 677 patients who had functional tricuspid regurgitation and left cardiac valve disease and underwent tricuspid valvuloplasty and left cardiac valve surgery were retrospectively. Among these patients, 353 underwent simple suture annuloplasty (group A) while the rest 324 patients underwent artificial plastic ring annuloplasty (group B). The two-year and more-than-two-year clinical and ultrasonocardiograph (UCG) follow-up data of the two groups were obtained and compared.Results:A total of 600 patients (88.6%) completed the long-term follow-up (more than two years). The two-year follow-up showed no significant difference in the incidence of mild tricuspid regurgitation between the two groups (82.2% vs. 92.7%, P=0.37). However, there were significantly more cases that developed into moderate to severe tricuspid regurgitation in group A than in group B (17.8% vs. 7.3%, P=0.031). The long-term follow-up revealed that the recurrence rate of tricuspid regurgitation in group B was significantly lower than that in group A (11.0% vs. 25.0%, P=0.029), and the ratio of cases developing into moderate to severe tricuspid regurgitation in group A was significantly higher than that in group B (28.9% vs. 9.9%, P=0.007). The comparison between the two intra-group time segments showed that the development of tricuspid regurgitation in group A was significantly increased (28.9% vs. 17.8%, P=0.022), but in group B it was relatively stable (9.9% vs. 7.3%, P=0.52).Conclusions:Artificial ring annuloplasty is associated with significantly less tricuspid regurgitation than simple suture annuloplasty.

Effect of Long-Term Systolic Blood Pressure Trajectory on Kidney Damage in the Diabetic Population: A Prospective Study in a Community-Based Chinese Cohort / 中华医学杂志(英文版)

<p><b>Background</b>Previous studies have shown that hypertension is an important factor contributing to the occurrence and progression of diabetic kidney damage. However, the relationship between the patterns of blood pressure (BP) trajectory and kidney damage in the diabetic population remains unclear. This prospective study investigated the effect of long-term systolic BP (SBP) trajectory on kidney damage in the diabetic population based on an 8-year follow-up community-based cohort.</p><p><b>Methods</b>This study included 4556 diabetic participants among 101,510 participants. BP, estimated glomerular filtration rate (eGFR), and urinary protein were measured every 2 years from 2006 to 2014. SBP trajectory was identified by the censored normal modeling. Five discrete SBP trajectories were identified according to SBP range and the changing pattern over time. Kidney damage was evaluated through eGFR and urinary protein value. A multivariate logistic regression model was used to analyze the influence of different SBP trajectory groups on kidney damage.</p><p><b>Results</b>We identified five discrete SBP trajectories: low-stable group (n = 864), moderate-stable group (n = 1980), moderate increasing group (n = 609), elevated decreasing group, (n = 679), and elevated stable group (n = 424). The detection rate of kidney damage in the low-stable group (SBP: 118-124 mmHg) was the lowest among the five groups. The detection rate of each kidney damage index was higher in the elevated stable group (SBP: 159-172 mmHg) compared with the low-stable group. For details, the gap was 4.14 (11.6% vs. 2.8%) in eGFR <60 ml·min·1.73 m and 3.66 (17.2% vs. 4.7%), 3.38 (25.0% vs. 7.4%), and 1.8 (10.6% vs. 5.9%) times in positive urinary protein, eGFR <60 ml·min·1.73 m and/or positive urinary protein, and eGFR decline ≥30%, respectively (P < 0.01).</p><p><b>Conclusion</b>An elevated stable SBP trajectory is an independent risk factor for kidney damage in the diabetic population.</p>

Clinical efficacy of two different tricuspid annuloplasty techniques in left cardiac valve surgery

Objective: To investigate the mid-and long-term clinical effects of tricuspid valvuloplasty with the implantation of an artificial plastic ring. Methods: Data of 677 patients who had functional tricuspid regurgitation and left cardiac valve disease and underwent tricuspid valvuloplasty and left cardiac valve surgery were retrospectively. Among these patients, 353 underwent simple suture annuloplasty (group A) while the rest 324 patients underwent artificial plastic ring annuloplasty (group B). The two-year and more-than-two-year clinical and ultrasonocardiograph (UCG) follow-up data of the two groups were obtained and compared. Results: A total of 600 patients (88.6%) completed the long-term follow-up (more than two years). The two-year follow-up showed no significant difference in the incidence of mild tricuspid regurgitation between the two groups (82.2% vs. 92.7%, P=0.37). However, there were significantly more cases that developed into moderate to severe tricuspid regurgitation in group A than in group B (17.8% vs. 7.3%, P=0.031). The long-term follow-up revealed that the recurrence rate of tricuspid regurgitation in group B was significantly lower than that in group A (11.0% vs. 25.0%, P=0.029), and the ratio of cases developing into moderate to severe tricuspid regurgitation in group A was significantly higher than that in group B (28.9% vs. 9.9%, P=0.007). The comparison between the two intra-group time segments showed that the development of tricuspid regurgitation in group A was significantly increased (28.9% vs. 17.8%, P=0.022), but in group B it was relatively stable (9.9% vs. 7.3%, P=0.52). Conclusions: Artificial ring annuloplasty is associated with significantly less tricuspid regurgitation than simple suture annuloplasty.