Assessment of Individual and Mixed Effects of Six Minerals on Thyroid Hormones in Chinese Pregnant Women.

The biosynthesis of thyroid hormones is essential for brain and neurological development. It requires iodine as a key component but is also influenced by other nutrients. Evidence for the combined nutrient status in relation to thyroid hormones during pregnancy is limited. We aimed to investigate the joint associations of iodine, selenium, zinc, calcium, magnesium and iron with maternal thyroid functions in 489 pregnant women from Hangzhou, China. Serum levels of six essential minerals and thyroid function parameters were measured during the first antenatal visit. Linear regression, quantile g-computation and Bayesian kernel machine regression were used to explore the individual and joint relationships between the six minerals and thyroid hormones. Linear regression analyses revealed that calcium was positively associated with free triiodothyronine (FT3). Zinc was positively associated with free thyroxine (FT4). Iodine was negatively associated with thyroid-stimulating hormone (TSH) and positively associated with FT3 and FT4. The quantile g-computation and BKMR models indicated that the joint nutrient concentration was negatively associated with TSH and positively associated with FT3 and FT4. Among the six minerals, iodine contributed most to thyroid function. The findings suggested that maintaining the appropriate concentration of minerals, either as individuals or a mixture, is important for thyroid health during pregnancy.

Ameliorative effects of silybin against avermectin-triggered carp spleen mitochondrial dysfunction and apoptosis through inhibition of PERK-ATF4-CHOP signaling pathway.

The aim of this study was to investigate the splenic tissue damage of environmental biological drug avermectin to freshwater cultured carp and to evaluate the effect of silybin on the splenic tissue damage of carp induced by avermectin. A total of 60 carp were divided into 4 groups with 15 carp in each group, including the control group fed with basic diet, experimental group fed with basal diet and exposed to avermectin (avermectin group), experimental group fed with basal diet supplement silybin (silybin group), and experimental group fed with basal diet supplement silybin and exposed to avermectin (silybin + avermectin group). The whole test period lasted for 30 days, and spleen tissue was collected for analysis. In this study, H&E staining, mitochondrial purification and membrane potential detection, ATP detection, DHE staining, biochemical tests, qPCR, immunohistochemistry, and apoptosis staining were used to evaluate the biological processes of spleen tissue injury, mitochondrial function, oxidative stress, apoptosis, and endoplasmic reticulum stress. The results show that silybin protected carp splenic tissue damage caused by chronic avermectin exposure, decreased mitochondrial membrane potential, decreased ATP content, ROS accumulation, oxidative stress, apoptosis, and endoplasmic reticulum stress. Silybin may ameliorate the splenic tissue damage of cultured freshwater carp caused by environmental biopesticide avermectin by alleviating mitochondrial dysfunction and inhibiting PERK-ATF4-CHOP-driven mitochondrial apoptosis. Adding silybin into the diet becomes a feasible strategy to resist the pollution of avermectin and provides a theoretical basis for creating a good living environment for freshwater carp.

Difenoconazole disrupts the blood-brain barrier and results in neurotoxicity in carp by inhibiting the Nrf2 pathway mediated ROS accumulation.

Excessive use of hard-to-degrade pesticides threatens the ecological health of aquatic systems. This study aimed to investigate difenoconazole (DFZ) residues in the environment induced neurotoxicity in carp and the underlying mechanisms. A total of thirty-six carps were divided into three groups and exposed to 0, 0.5, and 2.0 mg/L DFZ for 96 h, respectively. The alterations in behavior and blood-brain barrier (BBB) were examined, and potential mechanisms were explored using immunological assays and biochemical methods. The results showed that DFZ exposure caused behavioral freezing, reduced feeding, and neuronal necrosis in carp. Mechanistically, DFZ triggered ROS accumulation and destroyed the balance between oxidation and antioxidation with increased lipid peroxidation product MDA contents and reduced antioxidant enzymes SOD and CAT activities in the carp brain by inhibiting the NF-E2-related factor 2 (Nrf2) pathway. The activation of oxidative stress further reduced tight junction proteins and MMP levels, thereby destroying BBB and leading to DFZ leakage into the brain. Increased BBB permeability additionally led to DFZ activation of nuclear factor kappa-B signaling-mediated inflammatory cytokine storm, exacerbating neuroinflammation. Meanwhile, DFZ exposure activated mitochondria-associated apoptosis in the carp's brain by up-regulating Bcl-2 associated X protein, cleaved-caspase3, and cytochrome C and decreasing B-cell lymphoma-2 levels. Interestingly, the carp's brain initiated a protective autophagic response via the PI3K/AKT/TOR pathway intending to counteract the neurotoxicity of DFZ. Overall, we concluded that accumulation of DFZ at high concentrations in the aquatic systems disrupted the BBB and resulted in neurotoxicity in carp through inhibition of Nrf2 pathway-mediated ROS accumulation. This study provides a reference for monitoring DFZ residues in the environment and a new target for the treatment of DFZ-induced neurotoxicity in carp.

Controlled CRISPR-Cas9 Ribonucleoprotein Delivery for Sensitized Photothermal Therapy.

Manipulation of CRISPR delivery for stimuli-responsive gene editing is crucial for cancer therapeutics through maximizing efficacy and minimizing side-effects. However, realizing controlled gene editing for synergistic combination therapy remains a key challenge. Here, a near-infrared (NIR) light-triggered thermo-responsive copper sulfide (CuS) multifunctional nanotherapeutic platform is constructed to achieve controlled release of CRISPR-Cas9 ribonucleoprotein (RNP) and doxorubicin for tumor synergistic combination therapy involving in gene therapy, mild-photothermal therapy (PTT), and chemotherapy. The semiconductor CuS serves as a "photothermal converter" and can stably convert NIR light (808 nm) into local thermal effect to provide photothermal stimulation. The double-strand formed between CuS nanoparticle-linked DNA fragments and single-guide RNA is employed as a controlled element in response to photothermal stimulation for controlled gene editing and drug release. Hsp90α, one subunit of heat shock protein 90 (Hsp90), is targeted by Cas9 RNP to reduce tumor heat tolerance for enhanced mild-PTT effects (≈43 °C). Significant synergistic therapy efficacy can be observed by twice NIR light irradiation both in vitro and in vivo, compared to PTT alone. Overall, this exogenously controlled method provides a versatile strategy for controlled gene editing and drug release with potentially synergistic combination therapy.

Identification of active compound combination contributing to anti-inflammatory activity of Xiao-Cheng-Qi Decoction via human intestinal bacterial metabolism.

Human intestinal bacteria play an important role in the metabolism of herbal medicines, leading to the variations in their pharmacological profile. The present study aimed to investigate the metabolism of Xiao-Cheng-Qi decoction (XCQD) by human intestinal bacteria and to discover active component combination (ACC) contributing to the anti-inflammatory activity of XCQD. The water extract of XCQD was anaerobically incubated with human intestinal bacteria suspensions for 48 h at 37 °C. A liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) method was performed for identification of the metabolites. In addition, the anti-inflammatory effects of XCQD and biotransformed XCQD (XCQD-BT) were evaluated in vitro with cytokines in RAW264.7 cells induced by lipopolysaccharide (LPS). A total of 51 compounds were identified in XCQD and XCQD-BT. Among them, 20 metabolites were proven to be transformed by human intestinal bacteria. Significantly, a combination of 14 compounds was identified as ACC from XCQD-BT, which was as effective as XCQD in cell models of inflammation. In conclusion, this study provided an applicable method, based on intestinal bacterial metabolism, for identifying combinatory compounds responsible for a certain pharmacological activity of herbal medicines.

[Research progress of platelet activating factor(PAF) receptor antagonist].

Platelet activating factor(PAF), an endogenous synthesized phospholipid transmitter, has widely biological activities. It has "signal transmission" effect in various life processes, but abnormality of concentration in vivo will promote or aggravate the diseases, such as, cerebral ischemia, myocardial injury, multi-organ failure, asthma, injury of liver and kidney, severely affecting the normal life activities of body. In recent years, with the development of medical science and technology, more and more attention has been paid to the research of platelet activating factor receptor antagonist. Components of animals, plants, microbial fermentation, and synthetic composition all can reflect such activity. Ginkgolide B and cytopone are the most representative herbal compositions at present. This paper referred to the research status of platelet activating factor receptor antagonist in recent years, made a summary of the researches on biological effect of platelet activating factor and platelet receptor antagonist of different sources, so as to provide a reference for the exploration of effective and safe platelet activating factor receptor antagonists.

[Clinical observation of acupuncture at Zhiyang (GV 9) Bazhen points combined with pantoprazole for gastroesophageal reflux cough with damp-heat type].

OBJECTIVE: On the basic treatment, to observed the effect difference between acupuncture at Zhiyang (GV 9) Bazhen points combined with pantoprazole and simple pantoprazole for gastroesophageal reflux cough with damp-heat type. METHODS: A total of 102 patients were randomly assigned into an observation group and a control group, 51 cases in each group. The patients in the two groups were given domperidone tablets (10 mg each time, 3 times a day). The patients in the control group were treated with pantoprazole capsule (40 mg each time, once a day). On the basis of the control group, the patients in the observation group were treated with Zhiyang (GV 9) Bazhen points, once a day, 5 times a week. All the treatment was given for 8 weeks. The indexes were observed before and after treatment, including cough symptom at daytime and nighttime scores, TCM symptom (heartburn, acid regurgitation, chest pain, epigastric pain, throat discomfort) scores and quality of life scores of cough questionnaire in Leicester (physiological, psychological and social aspects). The clinical effects were compared. RESULTS: After treatment, the daytime and nighttime cough scores, TCM symptom (heartburn, acid regurgitation, chest pain, epigastric pain, throat discomfort) scores were lower, and the quality of life scores of cough questionnaire in Leicester (physiological, psychological and social aspects) were higher than those before treatment in the two groups (all P<0.05), with better results in the observation group (all P<0.05). The total effective rate of the observation group was 94.1% (48/51), which was better than 80.4% (41/51) of the control group (P<0.05). CONCLUSION: On the basic treatment, acupuncture at Zhiyang (GV 9) Bazhen points combined with pantoprazole can improve TCM symptoms, such as cough, of the patients with gastroesophageal reflux cough with damp-heat type, and improve their quality of life.

[Clinical observation on severe acute pancreatitis treated with electroacupuncture at Dachangshu (BL 25) and Shangjuxu (ST 37) combined with ulinastatin].

OBJECTIVE: To observe the clinical therapeutic effects on severe acute pancreatitis (SAP) treated with electroacupuncture (EA), sparse-dense wave and 2 Hz/15 Hz, at Dachangshu (BL 25) and Shangjuxu (ST 37) assisting ulinastetin and explore the effective therapeutic method for SAP. METHODS: A total of 120 patients of SAP were randomized into an observation group and a control group, 60 cases in each one. In the control group, the routine western medicine was adopted with the intravenous drip with ulinastatin. In the observation group, on the basis of the treatment as the control group, EA was applied at Dachangshu (BL 25) and Shangjuxu (ST 37). The treatment was given once every day, 20 min each time. The treatment was 2 weeks in the two groups. The recovery time of defecation, the recovery time of bowel sound, the remission time of abdominal pain and the hospitalization time were recorded in the patients of the two groups. The changes in the tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and amylase, as well as the scores in the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) were compared in the patients of the two groups before treatment, and on the 3rd, 8th and 14th days of treatment. The therapeutic effects, the cases of surgical transfer or the cases of the transfer of intensive care unit (ICU) were compared between the two groups. RESULTS: The recovery time of defecation, the recovery time of bowel sound and the remission time of abdominal pain, as well as the hospitalization time in the observation group were shorter than those in the control group (all P <0.05). The levels of TNF-α, IL-6 and amylase, as well as the scores of APACHE Ⅱ on the 3rd, 8th and 14th days of treatment were all reduced as compared with those before treatment in the two groups (all P <0.05). The results in the observation group were lower than those in the control group (all P <0.05). The total effective rate was 95.0% (57/60) in the observation group, better than 81.7% (49/60) in the control group (P <0.05). There were 1 case of surgical transfer and 0 case in ICU transfer in the observation group, 3 cases of surgical transfer and 2 cases in ICU transfer in the control group, indicating the significant differences between the two groups (both P <0.05). . CONCLUSION: EA at Dachangshu (BL 25) and Shangjuxu (ST 37) displays the satisfactory accessory therapeutic effects on SAP treated with ulinastatin.

The activity of Hou-Po-Da-Huang-Tang is improved through intestinal bacterial metabolism and Hou-Po-Da-Huang-Tang selectively stimulate the growth of intestinal bacteria associated with health.

Hou-Po-Da-Huang Tang (HPDHT) was used for the treatment of intestinal tract diseases in China. However, the underlying mechanisms via the intestinal bacteria remain largely unclear. Therefore, the aim of this study was to evaluate the metabolism of HPDHT by the human intestinal bacteria and its modulating effect on the intestinal bacteria. As a result, a total of 34 compounds were identified in HPDHT and transformed HPDHT (T-HPDHT). Among them, 12 metabolites were proved to be transformed by human intestinal bacteria. In vitro assays showed that T-HPDHT exhibited more significant elevation of free radical scavenging activity and suppression on the production of nitric oxide (NO) and TNF-α when comparing to HPDHT. Additionally, in vivo experiment confirmed that HPDHT significantly increased activity of superoxide dismutase (SOD), attenuated the malondialdehyde (MDA) and TNF-α levels in the conventional rats compared with that of pseudo germ-free (PGF) rats. In addition, HPDHT could significantly enhance the mean counts of Bifidobacterium and Lactobacillus and inhibit the growth of Clostridium, and Enterobacteriaceae, relative to controls. Due to the transformation of HPDHT being dependent on the bacterial strain, the effect of HPDHT on the selective growth of Bifidobacterium bifidum 29521 and Lactobacillus plantarum 8014 was evaluated. The kinetic parameters of microbial growth and prebiotic activity scores indicated that HPDHT could selectively stimulate the growth of the strains Bifidobacterium bifidum 29521 and Lactobacillus plantarum 8014. Taken together, metabolism of HPDHT by intestinal bacteria is a critical step towards the emergence of their anti-oxidation, anti-inflammation and prebiotic activities. This study provided valuable information for further pharmacological research on HPDHT.

Evaluation of a TPTX model induced by ischemia.

Parathyroidectomy (PTX), especially total parathyroidectomy (TPTX), is often recommended for severe secondary hyperparathyroidism (SHPT) if other medical treatments fail. Accurate identification and resection of parathyroid gland (PTG) tissue is the cornerstone of PTX. The establishment of a rat TPTX model would be beneficial for several applications but faces the same problems. In this experiment, we studied the mechanisms of ischemia for the accurate identification and excision of PTG tissue to establish TPTX rat models and to analyze the effects of surgical removal of PTG tissue as well as the effects of different types of water intake in rats on clinical indices. We found that the ischemia method had advantages when establishing a rat TPTX model. Removal of the PTG tissue resulted in significantly changed postoperative indices, and varying the types of water intake induced significant differences in these indices after removal of the PTG tissue. The absolute value of the difference between the serum calcium and phosphorus concentrations (|Ca-P|) accurately reflected the effect of removal of the PTG tissue and was superior to the calcium-phosphorus product (Ca × P); Ca × P accurately reflected the effect of varying the types of water intake in rats and was superior to the |Ca-P|.

[On the depth of needle insertion].

The statement on the depth of needle insertion is collected from Neiiing (nternal Classic) and Nanjing (Classic of Medical Problems) and the relationship of it is discussed with meridians, diseases, symptoms, location of sickness, qi and blood changes in human body, time, layers of needle insertion, reinforcing and reducing of acupuncture, etc. The factors are explored for the determination of the puncture depth, which provides the important guidance for the depth of insertion, needling sensation, reinforcing and reducing manipulations, acupuncture effect and the improvement of acupuncture efficacy. The complex and mystery of acupuncture technique focus on the control of insertion depth. In clinical practice, the increase of needle insertion depth may not induce the equal increase of efficacy. The depth of needle insertion is rather relevant with stimulation at the effective acupoint in the effective layer.

[Impact on the gait time cycle of ischemic stroke in the treatment with yin-yang respiratory reinforcing and reducing needling technique].

OBJECTIVE: To compare the difference in the efficacy on gait time cycle of ischemic stroke between yin-yang respiratory reinforcing and reducing needling technique (yin-yang needling) and the conventional acupuncture. METHODS: Sixty cases of ischemic stroke were randomized into a conventional acupuncture group and a yin-yang needling group, 30 cases in each one. The basic treatment (the control of blood pressure, blood sugar and blood lipid, the intravenous drops of ginkgo leaf extract and dipyridamole injection and vinpocetine injection) were applied in the two groups. Additionally, in the conventional acupuncture group, the acupoints of the Stomach Meridian of Foot-Yangming [Biguan (ST 31), Liangqiu (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39), etc.] were selected and stimulated with the even needling technique. In the yin-yang needling group, the acupoints of yin meridians such as Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9) and Sanyinjiao (SP 6), etc. and the acupoints of yang meridians such as Biguan (ST 31), Liangqiu (ST 34) and Yanglingquan (GB 34), etc. were selected. The reducing manipulation of respiratory reinforcing and reducing technique was applied to the acupoints of yin meridians and the reinforcing manipulation was applied to the acupoints of yang meridians. The kinematics time parameters were determined and compared before and 4 weeks after treatment. RESULTS: After treatment, the differences in the gait cycle, the phase time of standing (%), the phase time of single support (%), the phase time of unilateral sway (%) on the affected (healthy) foot and phase time of double support (%) were significant as compared with those before treatment in the patients of the two groups (all P < 0.05), in which, the gait cycle (1.75 +/- 0.21 vs 2.02 +/- 0.37), the phase time of standing (%) on the affected (healthy) foot [(65.41 +/- 5.20)% vs (68.37 +/- 6.24)%, (70.99 +/- 6.47)% vs (74.51 +/- 5.19)%], the phase time of unilateral sway (%) on the affected (healthy) foot [(36.08 +/- 4.86)% vs (33.65 +/- 2.94)%, (31.04 +/- 3.41)% vs (26.77 +/- 2.67)%] and the phase time of double support (%) [(36.91 +/- 5.10)% vs (41.22 +/- 5.39)%] in the yin-yang needling group were improved much obviously after treatment as compared with those in the conventional acupuncture group. The differences in support phase time (%), single support phase time (%) and sway phase time (%) were significant between the affected limb and healthy limb of the two groups after treatment (all P < 0.05). CONCLUSION: Yin-yang respiratory reinforcing and reducing needling technique effectively improves hemiplegic gait movement cycle and walking function in patients of ischemic stroke, which is superior to the conventional acupuncture treatment.

[Impacts of yin-yang meridians acupuncture with respiratory reinforcing and reducing manipulation on lower limbs balance function in stroke patients].

OBJECTIVE: To observe the impact on lower limbs balance function in treatment of yin-yang meridians acupuncture with respiratory reinforcing and reducing manipulation involved in the patients of stroke by applying B-PHY balance function test training system so as to provide the objective evidence in treatment of stroke; with acupuncture. METHODS: One hundred patients were randomized into an observation group and a control group, 50 cases in each one. In the control group, the basic treatment was applied, without other relevant rehabilitation therapies associated. In the observation group, with the basic treatment as the control group's, the therapy of the yin-yang meridians acupuncutre with respiratory reinforcing and reducing manipulation was adopted. On the yin meridians, Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9), Sanyinjiao (SP 6) and the others were selected and stimulated with reducing manipulation achieved by the coordination of patient's respiration. On the yang meridians, Biguan (ST 31), Liangqiu (ST 34), Yanglingquan (GB 34) and the others were selected and stimulated with reinforcing manipulation achieved by the coordination of patient's respiration. The treatment was given once a day and for 28 days totally. Before treatment and in 28 days of treatment, B-PHY balance function test training system was used to determine the weight shift track parameters (track length, peripheral square, track length of per unit square, left-right offset and rectangle square), the weight shift track distance parameters [mean of X axle weight shift distance (Mean-X), mean of Y axle weight shift distance (Mean-Y), maximum of X axle weight shift distance (Max-X), maximum of Y axle weight shift distance (Max-Y), weight shift distance (LSKG), weight shift square (SSKG), square ratio of weight shift (LFS)], stability coefficient (SI) and weight distribution coefficient (WDI). RESULTS: After treatment, the differences in the weight shift track parameters, SI and WDI were significant as compared with those before treatment in the patients of the two groups (all P<0.01); while the differences in the weight shift distance parameters in the observation group were improved obviously after treatment as compared with those before treatment (all P<0.01), the differences of Mean-X, Max-Y and LFS in the control group were improved after treatment as compared with those before treatment (all P<0.01). Except SSKG, the improvements after treatment in the rest indices in the observation group were better than those in the control group (all P<0.05). CONCLUSION: The yin-yang meridians acupuncture with respiratry reinforcing and re- ducing manipulation effectively improves the lower limbs balance function in the patients of stroke.

[Efficacy observation on wrist-ankle needle for primary dysmenorrhea in undergraduates].

OBJECTIVE: To compare efficacy difference among wrist-ankle needle, body-acupuncture and ibuprofen in the treatment of primary dysmenorrhea. METHODS: Ninety-five cases were randomly divided into a wrist-ankle needle group (32 cases), a body-acupuncture group (31 cases) and an ibuprofen group (32 cases). Acupunc- , ture at Lower 1 and Lower 2 area was applied in the wrist-ankle needle group. Acupuncture at Guanyuan (CV 4) and Sanyinjiao (SP 6) were applied in the body-acupuncture group. Ibuprofen sustained-release capsules were given for oral administration in the ibuprofen group. The treatment began 3 days before menses, once a day, until pain was relieved. One menstrual cycle was taken as a treatment course, continuously for 3 courses and efficacy were observed in three groups. The symptom score of dysmenorrhea and visual analogue scale (VAS) were used to assess pain severity before and after treatment. RESULTS: 1The efficacy differences in three groups were statistically significant (P<0.01), in which the total effective rate was 90. 0% (27/30) in the wrist-ankle needle group, 73.4% (22/30) in the body-acupuncture group and 46. 7% (14/30) in the ibuprofen group. 2 After the treatment, symptom score of dysmenorrhea and VAS were all obviously lower than that before the treatment in three groups (all P<0.01). Compared with ibuprofen group (7.12+/-2.70), after the treatment symptom score of dysmenorrhea in the wrist-ankle needle group (4.00+/-3.40) and body-acupuncture group (5. 53+/-2. 80) was obviously decreased (P<0.01, P<0.05), and VAS in the wrist-ankle needle group was significantly reduced (P<0.05). Compared with body-acupuncture group (5. 53+/-2.80), symptom score of dysmenorrhea in the wrist-ankle needle group (4.00+/- 3. 40) was obviously decreased (P<0. 05). CONCLUSION: The wrist-ankle needle has better effect than body acupuncture and ibuprofen on the treatment of primary dysmenorrhea, which could significantly improve dysmenorrhea symptoms.

[Discussion on the acupuncture and constitution according to the Miraculous Pivot].

The texts on constitution of Miraculous Pivot were analyzed and summarized according to literature study. The findings show that constitution is an important element in acupuncture treatment according to the texts of Miraculous Pivot. Different constitution generates the difference of yin-yang, deficiency or excess of qi and blood which is the important basis for syndrome differentiation of Miraculous Pivot. Therefore, the improvement in constitution is the way to increase the therapeutic effect of acupuncture.