This paper summarized Professor BU He's clinical experience in the integrative internal and external treatment of pediatric functional constipation. He believes that dysfunction of the spleen in transportation leads to impaired function of the sanjiao （三焦） and intestinal obstruction， which constitutes the core pathogenesis of pediatric functional constipation. The treatment emphasizes regulating the spleen and harmonizing the pivot， as well as unblocking the bowel and eliminating stagnation. An integrated approach combining internal and external therapies is advocated. Internally， Chinese herbal medicine， primarily Modified Sijunzi Decoction and Shaoyao Gancao Decoction （四君子汤合芍药甘草汤加减） is used to regulate the internal organs. Externally， pediatric tuina and moxibustion on qi interlocking are employed. Together， these methods fortify the spleen and facilitate transportation， restoring the smooth function of the sanjiao pivot， and ensuring intestinal patency， thereby relieving constipation.

ObjectiveTo explore the mechanism by which Zuoguiwan improves the pancreas development in the gestational diabetes mellitus （GDM） model by observing the effects of Zuoguiwan on the expression of key regulatory factors in different stages of pancreas development. MethodsPregnant Wistar rats were randomly assigned into blank， model， insulin detemir （20 U·kg^-1） and Zuoguiwan （1.89 g·kg^-1） groups （n=18）. GDM was induced by peritoneal injection of streptozotocin on day 6.5 （E6.5d） in the embryonic stage， and the blank group was given an equal volume of sodium citrate buffer. The modeling performance was assessed by measuring the blood glucose of pregnant rats. Except the blank group and model group， pregnant rats in other groups were administrated with corresponding drugs from E9.5d to delivery. The random blood glucose of pregnant rats was monitored， and the embryos and offspring rats were measured for the length and weighed on E12.5d， E18.5d and day 21 after birth （B21d）. The Lee's index of rats on B21d was calculated. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the fasting insulin （FINS） levels of B22d rats and the Homeostasis Model Assessment for Insulin Resistance （HOMA-IR） was calculated. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， total bilirubin （TBIL）， total cholesterol （CHO）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL）， and low-density lipoprotein cholesterol （LDL） in E18.5d pregnant rats and B22d offspring were determined. The pathological changes in the pancreas of E12.5d， E18.5d and B22d rats were observed by hematoxylin-eosin （HE） staining. Western blot was used to determine the protein levels of pancreatic duodenal homeobox 1 （Pdx1）， pancreas-specific transcription factor 1a （Ptf1a）， and sex-determining region Y-box protein 9 （Sox9） in the pancreas of E12.5d embryos， Pdx1， Nkx2 homeobox 2 （Nkx2.2）， and hairy and enhancer of split-1 （Hes1） in the pancreas of E18.5d embryos， and Pdx1， v-maf musculoaponeurotic fibrosarcoma oncogene homolog A （Mafa）， and NK transcription factor-related homeobox gene family 6 locus 1 （Nkx6.1） in the pancreas of B22d rats. ResultsCompared with the blank group， the model group showed elevated blood glucose levels in pregnant rats on B0d， E9.5d， E12.5d， E15.5d， and E18.5d （P<0.05， P<0.01）， decreased body weight and body length （P<0.01） and increased Lee's index in the offspring. In addition， the B22d offspring showed rising levels of FBG， FINS， HOMA-IR， AST， and TG （P<0.01）， a declined level of HDL （P<0.01）， and pancreatic acinous cells with edema and loose arrangement. The pregnant rats on E18.5d exhibited raised levels of ALT， AST， and TG （P<0.05， P<0.01） in the pancreas and a declined level of HDL （P<0.05）. The E12.5d embryos showed up-regulated protein levels of Pdx1， Sox9， and Ptf1a in the pancreas （P<0.01） and the E18.5d embryos exhibited down-regulated protein levels of Pdx1， Nkx2.2， and Hes1 in the pancreas （P<0.01）. The protein levels of Pdx1， Nkx6.1， and Mafa in the pancreas of B22d offspring were down-regulated （P<0.01）. Compared with the model group， the insulin group exhibited lowered blood glucose in pregnant rats on B0d， E15.5d， and E18.5d （P<0.05， P<0.01）. The offspring in all treatment groups showcased increased body weight and body length （P<0.01） and decreased Lee's index. The B22d offspring exhibited declined levels of FBG， FINS， and HOMA-IR in the insulin group （P<0.01） and lowered levels of FBG and HOMA-IR in the Zuoguiwan group （P<0.01）. The B22d offspring in all the treatment groups showed reduced levels of ALT， AST， TBIL， CHO， TG， and LDL， a raised level of HDL， and alleviated edema of pancreatic acinous cells. The pregnant rats on E18.5d demonstrated declined levels of TG and ALT （P<0.05， P<0.01） and an elevated level of HDL （P<0.05）. The pancreas of E12.5d embryos presented down-regulated protein levels of Pdx1 and Sox9 and an up-regulated protein level of Ptf1a in the insulin group （P<0.05）. The pancreas of E12.5d embryos in the Zuoguiwan group presented down-regulated protein levels of Pdx1， Sox9， and Ptf1a （P<0.01）. All the treatment groups showed up-regulated protein levels of Pdx1， Nkx2.2， and Hes1 in the pancreas of E18.5d embryos （P<0.01） and Pdx1， Nkx6.1， and Mafa in the pancreas of B22d embryos （P<0.05， P<0.01）. ConclusionZuoguiwan can promote the growth and development and ameliorate the pathological changes in the pancreas of the offspring of GDM model by regulating the expression of Pdx1 pathway-related regulatory factors in different stages of pancreas development.

OBJECTIVE: To reveal the gap between the evidence of systematic reviews (SRs) and clinical concerns by systematically summarizing the evidence on acupuncture and moxibustion for frozen shoulder and investigating the concerns and needs of clinicians in treatment with acupuncture and moxibustion for this disease. METHODS: The articles of SR and Meta-analysis on acupuncture and moxibustion for frozen shoulder were searched from CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase and Cochrane Library, starting from the inception of each database up to December 31st, 2022. Two researchers screened the articles and extracted data independently. Using AMSTAR-2, the methodological quality of the included studies was evaluated. Based on systematic reviews and expert discussion, a questionnaire on clinical concerns of acupuncture and moxibustion for frozen shoulder was developed and distributed to clinicians. The discrepancies between the evidence and clinical concerns were compared from 5 dimensions, including population, interventions, control measures, outcome indicators and review time points. RESULTS: The evidence gaps existed between SRs and clinical concerns. In the existing studies, the needs of personalized treatment were not fully considered in terms of different syndromes/patterns of frozen shoulder and stages of illness, the outcome indicators were not employed properly, the time for outcome measurement was vague, the control groups were set up outside of standardization, and the methodological quality was lower. CONCLUSION It is suggested that future studies should improve the quality of methodology, lay more consideration to different patient groups, optimize outcome indicators and standardize the setting of control groups, so as to better meet the needs of patients and achieve the best match between evidence and clinicians' needs.
Humans ; Acupuncture Therapy ; Bursitis/therapy* ; Evidence Gaps ; Moxibustion ; Systematic Reviews as Topic ; Meta-Analysis as Topic

The pharmaceutical industry faces challenges in quality digitization for complex multi-stage processes, especially in small-sample systems. Here, an intelligent quality prediction and diagnostic (IQPD) framework was developed and applied to Tong Ren Tang's Niuhuang Qingxin Pills, utilizing four years of data collected from four production units, covering the entire process from raw materials to finished products. In this framework, a novel path-enhanced double ensemble quality prediction model (PeDGAT) is proposed, which combines a graph attention network and path information to encode inter-unit long-range and sequential dependencies. Additionally, the double ensemble strategy enhances model stability in small samples. Compared to global traditional models, PeDGAT achieves state-of-the-art results, with an average improvement of 13.18% and 87.67% in prediction accuracy and stability on three indicators. Additionally, a more in-depth diagnostic model leveraging grey correlation analysis and expert knowledge reduces reliance on large samples, offering a panoramic view of attribute relationships across units and improving process transparency. Finally, the IQPD framework integrates into a Human-Cyber-Physical system, enabling faster decision-making and real-time quality adjustments for Tong Ren Tang's Niuhuang Qingxin Pills, a product with annual sales exceeding 100 million CNY. This facilitates the transition from experience-driven to data-driven manufacturing.

The central amygdala (CeA) is a crucial modulator of emotional, behavioral, and autonomic functions, including cardiovascular responses. Despite its importance, the specific circuit by which the CeA modulates blood pressure remains insufficiently explored. Our investigations demonstrate that photostimulation of GABAergic neurons in the centromedial amygdala (CeM^GABA), as opposed to those in the centrolateral amygdala (CeL), produces a depressor response in both anesthetized and freely-moving mice. In addition, activation of CeM^GABA axonal terminals projecting to the nucleus tractus solitarius (NTS) significantly reduces blood pressure. These CeM^GABA neurons form synaptic connections with NTS neurons, allowing for the modulation of cardiovascular responses by influencing the caudal or rostral ventrolateral medulla. Furthermore, CeM^GABA neurons targeting the NTS receive dense inputs from the CeL. Consequently, stimulation of CeM^GABA neurons elicits hypotension through the CeM-NTS circuit, offering deeper insights into the cardiovascular responses associated with emotions and behaviors.
Animals ; GABAergic Neurons/physiology* ; Male ; Central Amygdaloid Nucleus/physiopathology* ; Hypotension/physiopathology* ; Mice ; Blood Pressure/physiology* ; Mice, Inbred C57BL ; Solitary Nucleus/physiology* ; Photic Stimulation ; Neural Pathways/physiology*

Neutralizing antibodies have been designed to specifically target and bind to the receptor binding domain (RBD) of spike (S) protein to block severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus from attaching to angiotensin converting enzyme 2 (ACE2). This study reports a distinctive nanobody, designated as VHH21, that directly catalyzes the S-trimer into an irreversible transition state through postfusion conformational changes. Derived from camels immunized with multiple antigens, a set of nanobodies with high affinity for the S1 protein displays abilities to neutralize pseudovirion infections with a broad resistance to variants of concern of SARS-CoV-2, including SARS-CoV and BatRaTG13. Importantly, a super-resolution screening and analysis platform based on visual fluorescence probes was designed and applied to monitor single proteins and protein subunits. A spontaneously occurring dimeric form of VHH21 was obtained to rapidly destroy the S-trimer. Structural analysis via cryogenic electron microscopy revealed that VHH21 targets specific conserved epitopes on the S protein, distinct from the ACE2 binding site on the RBD, which destabilizes the fusion process. This research highlights the potential of VHH21 as an abzyme-like nanobody (nanoabzyme) possessing broad-spectrum binding capabilities and highly effective anti-viral properties and offers a promising strategy for combating coronavirus outbreaks.
Single-Domain Antibodies/immunology* ; Spike Glycoprotein, Coronavirus/metabolism* ; SARS-CoV-2/immunology* ; Animals ; Humans ; Antibodies, Neutralizing/immunology* ; Camelus ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Angiotensin-Converting Enzyme 2

Objective:To analyze the effect of nucleotide sequence variants in the 5′ untranslated region (5′UTR) of Helicobacter pylori (Hp) cagA on mRNA secondary structure, as well as its regulatory role in cytotoxin-associated gene A (CagA) protein expression and bacterial virulence. Methods:The upstream nucleotide sequence of cagA was amplified by polymerase chain reaction (PCR) from 37 Hp strains, and the PCR products were sequenced. MEGA 5.0 software and RNAfold prediction software were used to analyze the nucleotide sequence variants of cagA 5′UTR and the changes in mRNA secondary structure of this region, respectively. Western blot was used to detect the expression level of CagA protein in Hp strains, and the regulatory effect of cagA 5′UTR variants on the difference in CagA protein expression was analyzed. An Hp-infected AGS cell model was established to evaluate bacterial adhesion rate; quantitative PCR (qPCR) was used to analyze the mRNA transcription levels of interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α); enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion levels of IL-8 and TNF-α proteins. Results:Nucleotide sequence alignment of cagA 5′UTR from 37 Hp strains showed that sequence differences were mainly concentrated in the -53motif, -10motif, + 34motif, and + 86motif regions. mRNA secondary structure prediction analysis revealed three types based on the StemB stem-loop structure: type Ⅰ (no StemB stem-loop), type ⅡA (StemB stem-loop with 3-4 base partial pairing), and type ⅡB (StemB stem-loop with 5 base full pairing). Western blot analysis showed that the CagA protein expression level was the highest in type Ⅰ Hp strains (1.72±0.29) and the lowest in type ⅡB strains (0.81±0.26), with a statistically significant difference between the two types ( P=0.030). The adhesion rate of type Ⅰ Hp strains to AGS cells was (52.90±11.17)%, which was higher than that of type Ⅱ strains [(21.27±6.16)%]. qPCR results showed that the mRNA transcription levels of IL-8 and TNF-α in AGS cells induced by type Ⅰ Hp strains were higher than those induced by type Ⅱ strains (140.23±24.47 vs 76.16±8.76, P=0.069; 55.20±9.04 vs 21.26±6.16, P=0.036). ELISA analysis further indicated that the secretion levels of IL-8 and TNF-α proteins in AGS cells induced by type Ⅰ Hp strains were also higher than those induced by type Ⅱ strains [(344.66±62.62)pg/ml vs (302.13±66.27)pg/ml, P=0.665; (131.04±4.94)pg/ml vs (79.17±11.32)pg/ml, P=0.014]. Conclusions:The cagA 5′UTR region of Hp strains exhibits significant nucleotide sequence variants. Hp strains with no StemB stem-loop (type Ⅰ) in the mRNA secondary structure show significantly increased CagA protein expression and higher bacterial pathogenic potential.

Objective The application of 18F-PSMA-1007 PET/CT in the precise diagnosis and treatment of prostate cancer(PCa)is gradually increasing.This study aimed to explore the predictive value of 18F-PSMA-1007 PET/CT for positive surgical margins after radical prostatectomy(RP).Methods A total of 173 PCa patients who were pathologically confirmed and underwent RP in our hospital from July 2020 to August 2023 were retrospectively recruited.All patients underwent 18F-PSMA-1007 PET/CT whole-body imaging before surgery.According to the postoperative pathological results,they were divided into a negative surgical margin group with 83 cases and a positive surgical margin group with 90 cases.The differences in age,SUVmax,SUVmean,preoperative total prostate specific antigen(tPSA),prostate specific antigen density(PSAD),prostate volume,Gleason score,and clinical T staging between the groups were compared,and the risk factors were further analyzed by Logistic regression.Results The preoperative tPSA(Z=-3.252,P=0.001),SUVmax(Z=2.531,P=0.011),and clinical T staging(P=0.018)were all lower in the negative surgical margin group than in the positive surgical margin group.PSAD(OR=3.492,95％CI:1.095-11.133,P=0.035),SUVmax(OR=1.036,95％CI:1.003-1.070,P=0.034),and clinical T staging(OR=3.364,95％CI:1.117-10.133,P=0.045)were independent risk factors for positive surgical margins.Conclusion 18F-PSMA-1007 PET/CT has certain predictive value for positive surgical margins after RP in PCa patients.High PSAD,high SUVmax and high clinical T stage were independent risk factors for predicting positive margin after PR surgery,thus providing some reference for the clinical treatment strategy of PCa patients.

Objective To evaluate the efficacy of non-ablative vaginal erbium-doped yttrium aluminum garnet(Er:YAG)laser therapy in improving symptoms in women with mild-to-moderate stress urinary incontinence(SUI).Methods A prospective,multicenter,randomized controlled trial was conducted.A total of 126 female SUI patients were enrolled and randomized in a 2∶1 ratio to the treatment group(n=84,receiving Er:YAG laser therapy)and the sham control group(n=42,undergoing non-energized laser device with shielded beam).Efficacy outcomes were compared between the two groups,including the International Consultation on Incontinence Questionnaire-Short Form(ICI-Q-SF)scores at baseline and 1,3,6 months after treatment,response rate 3 month after treatment,urine leakage volume,daily incontinence episodes and visual analog scale(VAS).Results One month and 3 months after treatment,the ICI-Q-SF scores in the treatment group(8.07±3.33 and 7.04±3.04,respectively)were significantly lower than those in the control group(9.57±4.03 and 9.65±3.27,respectively;both P＜0.05).The 3-month response rate was significantly higher in the treatment group(71.43％)compared to the control group(35.71％,P＜0.05).Improvements in urine leakage volume 1 and 3 months after treatment were significantly greater in the treatment group versus the control group(both P＜0.05).A statistically significant reduction in daily incontinence episodes was observed in the treatment group 1 month after treatment compared to the control group[2(0,5)times vs.4(2,8)times,P＜0.05].VAS score in the treatment group markedly decreased 1 month after treatment,comparable to the level observed in the control group[0(0,2.45)vs.0(0,0),P＜0.05].Conclusion Non-ablative vaginal Er:YAG laser therapy significantly alleviates symptoms in women with mild-to-moderate SUI and represents a viable treatment option for this condition.

Objective:To observe the efficacy and safety of combined lienal polypeptide injection therapy in the treatment of Mycoplasma pneumoniae pneumonia（MPP）in children aged 3 to 14 years old in multiple clinical centers.Methods:A randomized，controlled，multi-center clinical study design was adopted.A total of 240 hospitalized children aged 3 to 14 years old with MPP from 7 hospitals from September 1，2023 to January 31，2024 were included.According to the severity of pneumonia，they were divided into the mild MPP group with 80 cases and the severe MPP/refractory MPP（SMPP/RMPP）group with 160 cases，and then randomly divided into the control group and the experimental group at a ratio of 1 ∶1，using the random number table method.After screening，subjects entered a treatment period of 5 to 7 days.The control group was treated with azithromycin，while the experimental group was treated with azithromycin plus lienal polypeptide injection .The recovery of lung CT，length of hospital stay，duration of fever，cough score，whether mild cases developed into severe or refractory cases，duration of hormone use，use of intravenous immunoglobulin（IVIG），bronchoscopy treatment，and immune function were observed between the two groups to evaluate the efficacy of lienal polypeptide injection.Adverse events after medication，vital signs，blood routine，urine routine，liver function，myocardial enzymes，renal function，and electrocardiogram were observed to evaluate the safety. Results:A total of 231 subjects have completed the trial in the 7 hospitals，including 118 cases in the experimental group and 113 cases in the control group.Main observation index：the rate of lung CT aggravation in the experimental group was lower than that in the control group（2.6% vs 15.3%， P<0.01），and the difference was statistically significant.Secondary indexes：there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.However，the rate of cases of plastic bronchitis（PB）found under bronchoscopy in the experimental group was lower than that in the control group（0 vs 18.8%， P=0.03），and the difference was statistically significant.Among the mild MPP（72 cases），there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and the improvement rate of lung CT between the two groups（all P>0.05）.However，compared with the control group，the rate of cases developing into SMPP/RMPP in the experimental group was less（24.3% vs 48.6%， P=0.03），and the difference in IgG before and after treatment was small［0.53（-0.04，1.18）g/L vs 1.33（0.48，2.25）g/L， P=0.01］.Among the SMPP/RMPP cases（159 cases），the rate of cases of PB found under bronchoscopy in the experimental group was less than that in the control group（0 vs 20%， P=0.04），and the rate of cases with aggravated lung CT in the experimental group was less than that in the control group（1.3% vs 19.5%， P<0.01），and the improvement rate of lung CT in the experimental group was higher than that in the control group（88.8% vs 75.3%， P=0.03），with statistically significant differences.There were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.Two cases in the experimental group developed rashes，which improved after the drug was discontinued.There were no serious adverse reactions such as abnormal vital signs like dyspnea and cyanosis due to the use of lienal polypeptide injection.There were no obvious changes in blood routine，liver function，myocardial enzymes，renal function，electrocardiogram，and urine routine values before and after medication compared with the baseline. Conclusion:The combined use of lienal polypeptide injection in the treatment of MPP in children can reduce the probability of the transformation from mild cases to SMPP/RMPP，reduce the rate of aggravation of the image findings，promote the absorption of lung inflammation，reduce the rate of PB found under bronchoscopy，and has good safety.