OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment of "biaoben acupoint combination" on cardiomyocyte mitochondrial fission in the rats with myocardial ischemia-reperfusion injury (MIRI) and explore its mechanism. METHODS: Fifty male SD rats were randomly divided into a sham-operation group, a model group, an EA pretreatment group, an EA pretreatment + Compound C group and an EA pretreatment+ML385 group, 10 rats in each group. In the EA pretreatment, the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, EA was delivered at bilateral "Neiguan" (PC6), "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, with continuous wave and 2 Hz of frequency, 1 mA of current, once daily for consecutive 7 days. On day 8, in the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, 30 min before model preparation, the intraperitoneal injection with Compound C (0.3 mg/kg) and ML385 (30 mg/kg) was administered respectively. Except in the sham-operation group, the ligation of the left anterior descending coronary artery was performed to prepare MIRI rat model in the rest groups. In the sham-operation group, the thread was not ligated. After modeling, the content of reactive oxygen species (ROS) in the ischemic area was measured by flow cytometry, superoxide dismutase (SOD) was detected using xanthine oxidase method, and malondialdelyde (MDA) was detected using thiobarbituric acid (TBA) chromatometry. The morphology of myocardial tissue in the ischemic area was observed with HE staining, and the mitochondria ultrastructure of cardiomyocytes observed under transmission electron microscopy. Using immunofluorescence analysis, the positive expression of mitochondrial fission factor (MFF), mitochondrial fission 1 protein antibody (Fis1) and dynamin-related protein 1 (Drp1) was detected; and with immunohistochemical method used, the protein expression of adenosine monophosphate-activated protein kinase (AMPK), nuclear factor E2-associated factor2 (Nrf2) and Drp1 in the ischemic area was detected. RESULTS: Compared with the sham-operation group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 increased in the model group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 decreased (P<0.01), and the protein expression of Drp1 elevated (P<0.01). Compared with the model group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01), and the protein expression of Drp1 declined (P<0.01); and in the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the positive expression of MFF, Fis1 and Drp1, and the protein expression of Drp1 were all reduced (P<0.01). When compared with the EA pretreatment + Compound C group and the EA pretreatment+ML385 group, the content of ROS and MDA in the myocardial tissue of the ischemic area, and the positive expression of MFF, Fis1 and Drp1 were dropped in the EA pretreatment group (P<0.01); the content of SOD and the protein expression of AMRK and Nrf2 rose (P<0.01, P<0.05), and the protein expression of Drp1 decreased (P<0.05). In comparison with the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group, the cardiac muscle fiber rupture, cell swelling and mitochondrial disorders were obviously alleviated in the EA pretreatment group. The morphological changes were similar among the model group, the EA pretreatment+Compound C group and the EA pretreatment+ML385 group. CONCLUSION Electroacupuncture pretreatment of "biaoben acupoint combination" attenuates myocardial injury in MIRI rats, probably through promoting the phosphorylation of AMPK and Nrf2, inhibiting the excessive mitochondrial fission induced by Drp1, and reducing mitochondrial dysfunction caused by mitochondrial fragmentation and vacuolation.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Myocardial Reperfusion Injury/physiopathology* ; Myocytes, Cardiac/cytology* ; Rats ; Acupuncture Points ; Mitochondrial Dynamics ; Humans ; Reactive Oxygen Species/metabolism* ; NF-E2-Related Factor 2/genetics* ; Superoxide Dismutase/metabolism*

OBJECTIVE: To investigate the effects of the combined Yiqi Huoxue Jiedu formula (YHJF) on intestinal microbiota in elderly patients with pulmonary-derived sepsis and identify potential microbial targets. METHODS: A prospective randomized double-blind controlled trial was conducted. Elderly patients with pulmonary infection-induced sepsis admitted to the emergency department of Guangdong Provincial Hospital of Traditional Chinese Medicine (TCM), intensive care unit (ICU) of Fangcun Hospital, and ICU of Daxuecheng Hospital, from November 2020 to October 2021 were enrolled and randomized into two groups. Both groups received conventional Western medicine treatment. The observation group additionally received YHJF (composed of 15 g of Panax ginseng, 9 g of Panax notoginseng, and 3 g of Rheum palmatum, dissolved in 50 mL warm water) orally or via nasogastric tube twice daily for 7 days; while the control group received a placebo. Clinical data and fresh fecal samples were collected before treatment and on days 5-7 of treatment. Intestinal microbiota diversity and structure were analyzed via 16S rDNA sequencing and bioinformatics [α diversity, β diversity, and linear discriminant analysis effect size (LEfSe)]. RESULTS: Fifty-five patients were included (29 in the control group, 26 in the observation group). There were no significantly differences in gender, age, comorbidities, and baseline sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation II (APACHE II), acute gastrointestinal injury (AGI) classification score, and gastrointestinal failure (GIF) score between the two groups. Compared to the control group, the observation group showed significantly lower serum procalcitonin, APACHE II score, and greater reduction in GIF score by day 7. Thirty fecal samples were collected pre-treatment (baseline group), 29 post-treatment from the control group, and 26 from the observation group. Gut microbiota α diversity analysis revealed that Simpson index in the observation group and control group were significantly decreased compared to the baseline group [0.75 (0.53, 0.91), 0.81 (0.32, 0.91) vs. 0.88 (0.87, 0.89), both P < 0.05], but there was no significantly difference between the observation group and the control group. There were no significantly differences in Chao1, Ace, and Shannon indices among three groups. β diversity analysis indicated that distinct microbiota structures among three groups (R² = 0.096, P = 0.026). Species difference analysis showed that, at the phylum level, Firmicutes (53.69%), Actinobacteria (16.23%), Proteobacteria (15.39%), and Bacteroidetes (9.57%) dominated, with no significant intergroup differences. At the genus level, 38 taxa showed significant differences. Compared to the control group, the observation group exhibited increased Erysipelatoclostridium (P = 0.014) and Faecalibacterium (P = 0.013), and decreased Bacteroides (P = 0.009), Bilophila (P = 0.005), Eggerthella (P = 0.002), and Collinsella (P = 0.043). LEfSe analysis highlighted Lactobacillus salivarius, Erysipelatoclostridium, Collinsella, Cloacibacillus, and Bacteroides as key discriminators. CONCLUSION YHJF combined with conventional therapy alters intestinal microbiota structure in patients with elderly pulmonary-derived sepsis, with Bacteroides, Erysipelatoclostridium, and Collinsella identified as potential microbial targets.
Humans ; Gastrointestinal Microbiome/drug effects* ; Drugs, Chinese Herbal/therapeutic use* ; Double-Blind Method ; Sepsis/drug therapy* ; Aged ; Prospective Studies ; RNA, Ribosomal, 16S/genetics* ; Male ; Female ; Panax notoginseng ; Rheum

Inducing durable and effective immunity against severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) via vaccination is essential to combat the current pandemic of coronavirus disease 2019 (COVID-19). It has been noticed that the strength of anti-COVID-19 vaccination-induced immunity fades over time, which calls for an additional vaccination regime, as known as booster immunization, to restore immunity among previously vaccinated populations. Here we report a pilot open-label trial of a third dose of BBIBP-CorV, an inactivated SARS-CoV-2 vaccine (Vero cell), on 136 participants aged between 18 to 63 years. Safety and immunogenicity in terms of neutralizing antibody titers and cytokine/chemokine responses were analyzed as the main endpoint until day 28. While systemic reactogenicity was either absent or mild, SARS-CoV-2-specific neutralizing antibody titers rapidly arose in all participants within 4 weeks, surpassing the peak antibody titers elicited by the initial two-dose immunization regime. Broad increases of cellular immunity-associated cytokines and chemokines were also detected in the majority of participants after the third vaccination. Furthermore, in an exploratory study, a newly developed recombinant protein vaccine, NVSI-06-08 (CHO Cells), was found to be safe and even more effective than BBIBP-CorV in eliciting humoral immune responses in BBIBP-CorV-primed individuals. Together, these results indicate that a third immunization schedule with either homologous or heterologous vaccine showed favorable safety profiles and restored potent SARS-CoV-2-specific immunity, providing support for further trials of booster vaccination in larger populations.
Adolescent ; Adult ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19/prevention & control* ; COVID-19 Vaccines/adverse effects* ; China ; Humans ; Immunogenicity, Vaccine ; Middle Aged ; SARS-CoV-2 ; Vaccination ; Young Adult

Objective:To study the risk factors of postoperative permanent hypoparathyroidism after total thyroidectomy.Methods:In this retrospective study ,the receiver operating characteristic (ROC) curve of postoperative 24 h parathyroid hormone(PTH) values was used to predict the risk of permanent hypoparathyroidism.The clinical and pathological features that related to postoperative permanent hypoparathyroidism were studied by χ2 test and multivariate Logistic regression analysis. Results:Eight hundred and eighty-nine patients were enrolled, the incidence of postoperative transient and permanent hypoparathyroidism was 33.3% and 4.0%, respectively. When 24 h PTH levels less than or equal to 5.84 pg/ml were used as cut off value, the sensitivity of the prediction of permanent hypoparathyroidism was 100%, the specificity was 72%, and the positive predictive value was 22.5%. The presence of parathyroid gland in the pathologic specimen, parathyroid autotransplantation and PTH≤5.84 pg/ml at 24 h after operation were statistically significant risk factor for permanent hypoparathyroidism in multivariate analysis( χ2=10.900, P=0.001; χ2=4.415, P=0.044; χ2=13.576, P=0.000). Group analysis found that the lower 24 h PTH the higher incidence of permanent hypoparathyroidism. Conclusions:For patients undergoing total thyroidectomy, when parathyroid gland is involved and/or the PTH value is low at 24 h after surgery, the occurrence of permanent hypoparathyroidism is high.

Objective: To study the mRNA expressions of various CD97 isoforms in colorectal carcinoma tissues and their clinical significances. Methods: A total of 50 colon cancer patients in the First Affiliated Hospital of Wenzhou Medical University from December 2013 to May 2014 and human colon cancer cell lines SW480 and SW620 were enrolled. The real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expressions of CD97 human epidermal growth factor (EGF) (1, 2, 5), CD97EGF (1, 2, 3, 5) and CD97EGF (1, 2, 3, 4, 5) in colon cancer tissues, adjacent tissues, normal colon tissues, SW480 cells and SW620 cells. The relationship between the mRNA expression of CD97EGF (1, 2, 5) and the clinicopathological factors was analyzed. Results: Compared with those low expressions in adjacent tissues and normal tissues, the mRNA expressions of CD97 isoforms CD97EGF (1, 2, 5), CD97EGF (1, 2, 3, 5) and CD97EGF (1, 2, 3, 4, 5) in cancer tissues were highest, and the differences were statistically significant (0.71±0.20 vs. 0.40±0.09 vs. 0.35±0.07, F = 107.642, P < 0.01; 0.45±0.11 vs. 0.26±0.05 vs. 0.27±0.06, F = 94.231, P < 0.01; 0.41±0.10 vs. 0.21±0.05 vs. 0.19±0.03, F = 165.672, P < 0.01). In addition, the mRNA expression of CD97EGF (1, 2, 5) in colon cancer patients was associated with tumor infiltration depth (T₁-T₂ and T₃-T₄), clinical stages (Ⅰ-Ⅱ and Ⅲ-Ⅳ), and the differences were statistically significant (t = -2.582, P = 0.013; t = -5.062, P < 0.01). The mRNA expression of CD97EGF (1, 2, 5) in SW620 cells was higher than that in SW480 cells. Conclusions CD97 isoforms are highly expressed in colon cancer tissues, and CD97EGF (1, 2, 5) may play an important role in the development and invasion of colon cancer. The CD97 isoforms may be new markers in the treatment of colon cancer.

Objective: To analyze the diagnostic value of ultrasound guided 14 gauge coreneedle biopsy (US-CNB) in breast nodules. Methods: We retrospectively analyzed the pathological results of US-CNB and surgical excision from 373 breast nodules in Peking University International Hospital from Sep 2016 to Nov 2018 to evaluate the accuracy of 14g US-CNB. Results: A total of 349 patients(373 nodules)underwent US-CNB. US-CNB reported 282 benign lesions(75.6%, 282/373), 20 high-risklesions(5.4%, 20/373), and 71 malignant lesions(19.0%, 71/373). For 282 CNB reported benign lesions, the surgical pathology confirmed 235 lesions , 46 for high-risk lesions and 1 for malignant lesion with a concordancy of 83.3%(235/282)and the underestimation rate was 16.7%(47/282). US-CNB identified 20 high-risk lesions. According to surgical results, 15 were high-risk lesions and 5 were malignant lesions with a concordancy of 75% (15/20)and the underestimation rate was 25%(5/20). When it comes to malignant lesions, the excision results showed that 70 were malignant lesions and 1was high-risk lesion with a concordancy of 98.6%(70/71)and the overestimation rate was 1.4%(1/71). The concordance of the histological type , calculated for 50 invasive carcinomas, was 92% (46/50) with a kappa value of 0.77.The concordance of the histological grade could be calculated for 38 invasive ductal carcinomas with the Elston-Elllis Method . It was 89.5% (34/38) with a kappa value of 0.57. Conclusions The pathology result of 14gUS-CNB is in good consistency with surgical excision for breast benign and malignant lesions.

Objective: To study the diagnostic value of a multi-gene molecular testing in cytologically indeterminate thyroid nodules. Methods: From February 2018 to September 2018, patients with thyroid nodules who underwent fine needle aspiration(FNA) at Peking University Cancer Hospital were enrolled. Three hundred and sixty patients were included, consisting of 86 men and 274 women, with a mean age of 45.8 years (between 13 and 89 years old). Among 391 nodules, 141 were cytologically inderminate and 75 were resected. FNA samples underwent prospective testing using a next-generation sequencing (NGS) assay, which included 16 genes for point mutations and 26 types of gene fusions. The testing results of indeterminate nodules were compared with surgical outcomes, to determine the diagnostic performance. The results were compared with the BRAF V600E single gene mutation analysis by χ² test. Results: The multi-gene testing showed a sensitivity of 73.2%, specificity of 96.8%, positive predictive value of 96.8%, and negative predictive value of 73.2%. The diagnostic accuracy of multi-gene testing was significantly higher than the BRAF V600E mutation test (83.3% vs 73.6%, χ²=31.588, P<0.01). Conclusion Multi-gene testing in FNA samples is an effective method to diagnose cytologically indeterminate thyroid nodules, which has a higher accuracy than BRAF V600E mutation detection.

Objective: To investigate the diagnostic performance of fine-needle aspiration (FNA) cytology for the detection of lateral lymph node metastases in patients with papillary thyroid carcinoma (PTC). Methods: A total of 109 lateral lymph nodes with suspicious metastases from 85 patients were retrospectively subjected to FNAC, fine-needle aspiration thyroglobulin measurement (FNATg), and FNATg/SerumTg measurement. Lymph node pathological results after surgery were taken as the gold standard. Using Mann-Whitney U test, Pearson linear model and ROC curve were used for data analysis. Results: The sensitivity, specificity and accuracy of FNATg for the diagnosis of lateral neck lymph node metastasis were respectively 93.7%, 90.0% and 93.3% and those of FNATg/SerumTg were respectively 89.9%, 90.0% and 93.2% respectively, the threshold values for FNATg and FNATg/SerumTg were 0.925 ng/ml and 1.007, respectively. The sensitivity, specificity and accuracy of FNATg combined with FNAC were respectively 91.0%,93.5% and 94.4%. The existence of thyroid tissue and the expression of serum Tg did not affect the expression of lymph node FNATg. The FNATg cutoff value of 0.925 ng/ml showed the best diagnostic performance in patients with a thyroid gland, while the FNATg/SerumTg cutoff ratio of 14.95 showed the best diagnostic performance in patients without a thyroid gland. The serum TgAb significantly interfered with the expression of FNATg in the lateral neck metastatic lymph nodes (P=0.049). Conclusions FNATg alone or the combination of FNATg with FNAC are highly reliable in the diagnosis of lateral neck lymph node metastases in patients with PTC. The expression of TgAb may interfere with the accuracy of the diagnostic performance of FNATg.

OBJECTIVETo observe the expression change of mitophagy-related proteins in skeletal muscle in rats with spleen deficiency syndrome and to explain the partial action mechanism of acupuncture at Zusanli (ST 36) for spleen deficiency syndrome.

METHODSForty male SD rats, after normal feeding, were randomly divided into a normal group, a spleen deficiency group, a Zusanli group and a non-acupoint group, ten rats in each group. Except the normal group, the three factors modeling method was used for 14 days to establish the model of spleen deficiency syndrome on the other 3 groups. The rats in the Zusanli group were treated with EA at bilateral "Zusanli" (ST 36), while the rats in the non-acupoint group were treated with EA at bilateral non acupoint (dense-sparse wave, frequency of 2 Hz/100 Hz, 20 min per treatment, once a day for 10 days). The rats in the normal group and spleen deficiency group were treated with immobilization for 20 min per day, and no EA was given. The HPLC method was applied to measure the content of adenosine triphosphate (ATP) and adenosine monophosphate (AMP) in skeletal muscle. The Western blotting method was applied to measure the expression of adenosine monophosphate activated protein kinase (AMPK), p-AMPK, ULK1, p-ULK1，LC3-Ⅰand LC3-Ⅱ in skeletal muscle.

RESULTSThe ATP content in the spleen deficiency group was significantly lower than that in the normal group (<0.01); the ATP content in the Zusanli group was significantly higher than that in the spleen deficiency group (<0.05) but lower than that in the normal group (<0.05), there was no significant difference between the non-acupoint group and the spleen deficiency group (>0.05). Compared with the normal group, the AMP/ATP in the spleen deficiency group and the Zusanli group were significantly up-regulated (<0.01, <0.05). The differences of p-AMPK/AMPK between the spleen deficiency group and the normal group was not significant (>0.05). Compared with the normal group and spleen deficiency group, the p-AMPK/AMPK in the Zusanli group was significantly up-regulated (both <0.05). The p-ULK1/ULK1 and LC3-Ⅱ/LC3-Ⅰin the Zusanli group was higher than those in the normal group and spleen deficiency group (all <0.01).

CONCLUSIONEA at "Zusanli" (ST 36) might activate AMPK and produce stable ULK1/AMPK compound and increase the mitochondrial autophagy, which could regulate spleen-stomach and treat spleen deficiency.

Objective To explore the distribution characteristics of plasma renin concentration in patients with hypertension and the possibly methodological problems.Methods The subjects including 361 patients with hypertension[male: 184 cases, average age: (45.16±13.74)years old;female: 177 cases, average age: (51.04±12.68) years old]and 187 apparent healthy individuals[male: 92 cases, average age: (46.74±13.17)years old;female: 95 cases, average age: (47.33±13.18) years old]were recruited from Departments of Healthy Check-up and outpatients for hypertension in Tangshan Gongren Hospital. The plasma renin concentration was detected by chemiluminescence-Immunoassay.Results The plasma renin concentration shows log-transformed normal distribution both in healthy group and hypertension group. The range of plasma renin concentration in hypertension group is from 0.05 to 574.07 pg/ml, while that in apparent healthy group is from 3.24 to 120.40 pg/ml. The plasma renin concentration in both groups is higher in male than female (Hypertension t=2.19,P=0.029;Healthy people t=2.85,P=0.005). The average concentration of plasma renin in hypertension group is slightly higher, and the width of density distribution is larger in comparison with healthy group although there is no significant difference between them. However, the percentage of plasma renin abnormality was 26.59% (96/361) in hypertension group with 13.85%(50/361)of low renin subtype and 12.74%(46/361)of high renin subtype ConclusionsThe plasma renin concentration measured by Chemiluminescence-Immunoassay can be used as an effective tool for hypertension screening.