Detection of Carbamazepine and Its Metabolites in Blood Samples by LC-MS/MS / 法医学杂志

OBJECTIVES@#To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS).@*METHODS@#The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (Vacetonitrile∶Vmethanol=2∶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring.@*RESULTS@#The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 μg/mL and 252.14 ng/mL, respectively.@*CONCLUSIONS@#This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.

Effect of procalcitonin on lipopolysaccharide-induced expression of nucleotide-binding oligomerization domain-like receptor protein 3 and caspase-1 in human umbilical vein endothelial cells / 中国当代儿科杂志

OBJECTIVES@#To study the effect of procalcitonin (PCT) on lipopolysaccharide (LPS)-induced expression of the pyroptosis-related proteins nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 in human umbilical vein endothelial cells (HUVECs).@*METHODS@#HUVECs were induced by LPS to establish a model of sepsis-induced inflammatory endothelial cell injury. The experiment was divided into two parts. In the first part, HUVECs were randomly divided into four groups: normal control, LPS (1 μg/mL), PCT (10 ng/mL), and LPS+PCT (n=3 each). In the second part, HUVECs were randomly grouped: normal control, LPS, and LPS+PCT of different concentrations (0.1, 1, 10, and 100 ng/mL) (n=3 each). Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of NLRP3 and caspase-1 in each group.@*RESULTS@#In the first experiment: compared with the normal control group, the PCT, LPS, and LPS+PCT groups had significantly upregulated mRNA and protein expression levels of NLRP3 and caspase-1 (P<0.05); compared with the LPS group, the LPS+PCT group had significantly downregulated mRNA and protein expression levels of NLRP3 and caspase-1 (P<0.05). In the second experiment: compared with those in the LPS group, the mRNA and protein expression levels of NLRP3 and caspase-1 in the LPS+PCT of different concentrations groups were significantly downregulated in a concentration-dependent manner (P<0.05).@*CONCLUSIONS@#LPS can promote the expression of the pyroptosis-related proteins NLRP3 and caspase-1 in HUVECs, while PCT can inhibit the LPS-induced expression of the pyroptosis-related proteins NLRP3 and caspase-1 in HUVECs in a concentration-dependent manner.

A novel homozygous frameshift variant in DNAH8 causes multiple morphological abnormalities of the sperm flagella in a consanguineous Pakistani family / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.

A recurrent homozygous missense mutation in CCDC103 causes asthenoteratozoospermia due to disorganized dynein arms / 亚洲男科学杂志(英文版)

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.

New discovery and short-term effect analysis of tensor tympani muscle tenotomy for Meniere's disease under otoscope / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the feasibility and short-term effect of tensor tympani muscle Tenotomy in the treatment of Meniere's disease under otoscope. The possible pathogenesis was discussed and our views were put forward. Methods: The clinical data of 9 cases of Meniere's disease treated by otoscopic Tenotomy were analyzed retrospectively, including 2 males, 7 females, 5 right ones, 2 left ones and 2 bilateral ones. The average age was (56.33± 10.56) years, ranging from 38 to 75 years. We evaluated intraoperative findings and short-term postoperative efficacy, respectively evaluated postoperative aural fullness, tinnitus and hearing recovery, and evaluated postoperative vertigo attack in a short time. Results: Nine patients were completed the operation under general anaesthesia and otoscopy, and no serious complications occurred. We found new pathological changes in tympanic cavity in some cases during operation. There were rupture of round window membrane in 1 case, severe fibrous hyperplasia near the round window membrane and vestibular window and adhesion with ossicular chain in 1 case, fibrous cord and membranous hyperplasia near vestibular window and round window membrane in 1 case, fibrous hyperplasia and adhesion near the round window membrane in 2 cases, membranous hyperplasia and adhesion around vestibular window in 1 case. No fibrous hyperplasia was found in 3 cases in the tympanic cavity. The round window membrane can be exposed in 4 cases and failed in 5 cases. After 3 months of follow-up, we found that we found that 5/5 cases of aural fullness disappeared, 2/2 cases of earache disappeared, 3/8 cases of tinnitus improved, 5/8 cases presented with improvement and no aggravation, 3/3 cases of hearing allergy improved, 4/9 cases of hearing improved, and 5/9 cases showed no improvement or decrease. 9 patients were followed up for 3 months, of whom 8 patients had no vertigo, one patient suffered from vertigo twice within 3 months after operation, and the patient suffered from rupture of round window membrane. Conclusions: Endoscopic Tenotomy for Meniere's disease has obvious curative effect and quick recovery after operation. During the operation, we find that most of Meniere's patients have fibrous cord hyperplasia near the inner ear window membrane, which may be the pathological manifestation after repeated rupture and repair of the inner ear window membrane. The vertigo of Meniere's disease may be related to the destruction and repair of inner ear membrane structure caused by improper contraction or spasm of tympanic tensor muscle.

Design of non-metallic crown for primary molars and analyzation of stress distribution: a finite element study / 中华口腔医学杂志

Objectives: To study the design of nonmetallic crowns for deciduous molars by means of computer aided design and to analyze the key parameters of the nonmetallic crowns of deciduous molars using finite element method. Methods: The three-dimensional model of a mandibular second primary molar was constructed by using a micro-CT system. The thickness of the crown was limited to 0.5 mm and four different crown shapes (chamfer+anatomic, chamfer+non-anatomic, knife edge+anatomic and knife edge+non-anatomic) were designed. Then, the crown shape was limited as chamfer+non-anatomic and five different thicknesses of the crown (0.50, 0.75, 1.00, 1.25, 1.50 mm) were designed, and three different materials, including polyetherketoneketone (PEKK), polymethylmethacrylate (PMMA) resin and resin-infiltrated ceramic, were applied to make the crown. Stress distribution and fatigue of each component of the model under vertical and oblique loadings were analyzed by using finite element method. Non-axial retention analysis was performed on chamfer+non-anatomic crowns, made of PMMA resin, with thicknesses of 0.50, 0.75, 1.00, 1.25 and 1.50 mm. Results: Among the four crown shape designs, the chamfer+non-anatomic type crown showed the lowest von Mises stress and the highest safety factor. By comparing three different materials, the resin-infiltrated ceramic group showed obvious stress concentration on the buccal edge of the crown and the PEKK group showed stress concentration in the adhesive layer. Results of non-axial retention analysis showed that the torques required by the crowns with five thicknesses at the same rotation angle were as follows: 4 856.1, 4 038.1, 3 497.3, 3 256.3 and 3 074.3 N⋅m, respectively. The comparison of areas of the adhesives fracture among groups were as follows: 0.5 mm group < 0.75 mm group < 1.00 mm group < 1.25 mm group < 1.50 mm group. Conclusions: In the design of nonmetallic crowns for primary molars, the edge of the crown should be designed as chamfer, the shape of the inner crown should be non-anatomical and the minimum preparation amount of the occlusal surface should be 1.00 mm. Among the three materials, PMMA resin, of which elastic modulus is similar to the dentin and the dental adhesive, might be the most suitable material for the crowns of primary molars.

Effect of laparoscopic surgery for pancreatic cancer after neoadjuvant chemotherapy / 中华外科杂志

Objective: To evaluate the safety and feasibility of laparoscopic surgery after neoadjuvant chemotherapy for pancreatic cancer. Methods: Clinical data of 8 patients underwent laparoscopic surgery after neoadjuvant chemotherapy for pancreatic cancer at Fudan University Shanghai Cancer Center from September 2019 to June 2020 were reviewed retrospectively. There were 5 males and 3 females,aged from 47 to 72 years old. All patients underwent abdominal enhanced CT and PET-CT before operation to accurately evaluate the tumor stage and exclude distant metastasis. Results: Neoadjuvant chemotherapy with AG regimen(gemcitabine 1 000 mg/m2 and albumin bound paclitaxel 125 mg/m2) was received for 2 to 6 cycles before surgery. All 8 patients successfully completed the operation,including 5 cases of pancreaticoduodenectomy,2 cases of radical antegrade modular pancreatosplenectomy(RAMPS),and 1 case of total pancreatectomy. No conversion to laparotomy or laparoscopic assisted surgery. The operation time was 240 to 450 minutes,the blood loss was 100 to 500 ml,the postoperative length of stay was 10 to 16 days. During the follow-up period up to December 31, 2020, there was 1 case suffered grade B pancreatic leakage and abdominal infection. The numbers of resected lymph nodes were 9 to 31. All patients received R0 resection. The follow-up times were 4.5 to 9.5 months. One patient underwent RAMPS was diagnosed as liver metastasis after 2 months of the operation,and the other 7 patients still survived without tumor recurrence. Conclusion: Minimally invasive surgery of pancreatic cancer after neoadjuvant chemotherapy is safe and feasible in experienced pancreatic minimally invasive centers.

Screening of reference genes for quantitative real-time PCR in Artemisia argyi / 中国中药杂志

Artemisia Argyi Folium, a traditional Chinese medicine of important medicinal and economic value, sees increasing demand in medicinal and moxibustion product market. Screening stable and reliable reference genes for quantitative real-time PCR(qRT-PCR) is a prerequisite for the analysis of gene expression in Artemisia argyi. In this study, eight commonly used reference genes, Actin, 18s, EF-1α, GAPDH, SAND, PAL, TUA, and TUB, from the transcriptome of A. argyi, were selected as candidate genes. The expression of each gene in different tissues(roots, stems, and leaves) of A. argyi and in leaves of A. argyi after treatment with methyl jasmonate(MeJA) for different time(0, 4, 8, 12 h) was detected by qRT-PCR. Then, geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder were employed to evaluate their expression stability. The results demonstrated that Actin was the most stable reference gene in different tissues and in leaves treated with MeJA, and coming in the second was SAND. Furthermore, the expression of DXS and MCT which are involved in terpenoid backbone biosynthesis was detected in different tissues and after MeJA treatment. The results showed that the expression patterns of DXS and MCT in different tissues and under MeJA treatment calculated with Actin and SAND as internal reference genes were consistent, which validated the screening results. In conclusion, Actin is the most suitable reference gene for the analysis of gene expression in different tissues of A. argyi and after MeJA treatment. This study provides valuable information for gene expression analysis in A. argyi and lays a foundation for further research on molecular mechanism of quality formation of Artemisia Argyi Folium.

Research Progress of Signaling Pathways Related to Treatment of Colorectal Cancer with Chinese Medicine： A Review / 中国实验方剂学杂志

The occurrence and development of colorectal cancer as a complex disease involves the abnormality of multiple signaling pathways. Chinese medicine regulates a variety of biological processes such as tumor cell differentiation， cell proliferation， apoptosis， cell metastasis， cell cycle， and tumor angiogenesis to prevent the occurrence of colorectal cancer （inflammation-cancer transformation）， tumor metastasis （common metastases of colorectal cancer include liver metastasis， lung metastasis， bone metastasis， and lymphatic metastasis）， and multidrug resistance induced by chemotherapy， treat primary tumors， and mitigate the toxic and side effects of chemotherapy. The pathways of Chinese medicine in the treatment of colorectal cancer have been intensively studied. The available studies have demonstrated that Patrinia villosa Juss and Pien Tze Huang can regulate the Notch pathway to inhibit the growth of colorectal cancer cells. Curcumin and Quyu Jiedu decoction regulate Hippo pathway to inhibit the survival， proliferation， invasion， and migration of colorectal cancer cells. Kujin tea and luteolin suppress the proliferation of colorectal cancer cells and protect intestinal barrier by regulating Kelch-like epichlorohydrin-associated protein-1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/antioxidant response element （ARE） pathway. Icariin and ginkgolide C can regulate hepatocyte growth factor （HGF）/cellular-mesenchymal to epithelial transition factor （c-Met） pathway to induce apoptosis of colorectal cancer cells and prevent liver metastasis of colorectal cancer. Verbascoside and apigenin regulate p53 protein to promote apoptosis of colorectal cancer cells， reverse thymidylate synthase （TS）， and alleviate the multidrug resistance of colorectal cancer. Resveratrol and lycopene regulate insulin-like growth factor （IGF）/insulin-like growth factor receptor 1 （IGF1R） pathway to inhibit cancer cell metastasis and prolong disease-free survival. Cordycepin and Galla Chinensis water extract activate AMP-activated protein kinase （AMPK） pathway to inhibit the migration and invasion of cancer cells as well as the lung metastasis of colorectal cancer. The above summary aims to provide reference for the in-depth research on the treatment of colorectal cancer with Chinese medicine and inspire new research ideas.

Effect and Mechanism of "Liver-Soothing and Spleen-Invigorating" Therapies Against Liver Injury in Rats： An Exploration Based on TGR5 and Intestinal Mucosal Barrier Function / 中国实验方剂学杂志

ObjectiveTo explore the effects of different treatment methods of "soothing liver， invigorating spleen， soothing liver and invigorating spleen， soothing liver first and then soothing liver and invigorating spleen， as well as invigorating spleen first and then soothing liver and invigorating spleen" on liver depression combined with liver injury in rats and their action mechanisms. MethodA six-week rat model of liver depression combined with liver injury was established by restraint stress and subcutaneous injection of carbon tetrachloride （CCl4， 5.89 g·kg-1， once every three days）. At the same time， the drugs were given by gavage. Forty-eight male SD rats of clean grade were randomly divided into eight groups， namely the normal group， model group， bicyclol （0.2 g·kg-1） group， Sinisan （4.32 g·kg-1） group， Liu Junzitang （9.26 g·kg-1） group， Chaishao Liu Junzitang A （Chai A， soothing liver and invigorating spleen，13.57 g·kg-1） group， Chaishao Liu Junzitang B （Chai B， soothing liver first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， and Chaishao Liu Junzitang C （Chai C， invigorating spleen first and then soothing liver and invigorating spleen， 13.57 g·kg-1） group， with six rats in each group. The pathological changes in liver and colon tissues of each group were observed under light microscope and electron microscope. The serum biochemical indexes of the liver were detected using an automatic biochemical analyzer. The relative mRNA expression levels of Takeda G protein-coupled receptor 5 （TGR5） and intestinal mucosal zona occluden-1 （ZO-1）， Occludin， and Claudin-1 in the liver and colon were detected by reverse-transcription polymerase chain reaction （RT-PCR）. The positive expression rate of proliferating cell nuclear antigen （PCNA） in the colon was detected by immunohistochemistry. ResultCompared with normal group， the model group exhibited significantly elevated serum alkaline phosphatase （ALP）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， and direct bilirubin （DBIL） （P<0.01）， lowered TGR5 mRNA expression in liver tissue， up-regulated TGR5 mRNA expression in the colon tissue （P<0.05，P<0.01）， and down-regulated ZO-1， Occludin， and tight junction protein-1 (Claudin-1) mRNA expression and PCNA in the colon tissue （P<0.01）. Compared with the model group， bicyclol and Chai C remarkably decreased the levels of serum ALP， ALT， AST， TBIL， and DBIL （P<0.05，P<0.01）， while Liu Junzitang， Chai A， Chai B， and Chai C significantly up-regulated the TGR5 mRNA expression in the liver and down-regulated its expression in the colon （P<0.01）. Bicyclol， Chai A， Chai B， and Chai C enhanced the ZO-1 and Claudin-1 mRNA expression in the colon （P<0.05，P<0.01）. Bicyclol， Sinisan， and Chai C increased PCNA expression （P<0.01）. The comparison with the Chai C group showed that the TGR5 mRNA expression in the liver and ZO-1 mRNA expression in the colon of the bicyclol and Sinisan groups were lower， whereas the TGR5 mRNA expression in the colon was higher （P<0.01）. However， the PCNA expression in the colon of the Liu Junzitang and Chai B groups declined significantly （P<0.05）. ConclusionIn the presence of liver injury， invigorating spleen first helps to relieve the liver injury， and the efficacy of "spleen-invigorating" therapy in increasing the intestinal mucosal tight junction proteins and improving the gastrointestinal function is related to its activation of TGR5 to improve the intestinal mucosal barrier function， promote the renewal of intestinal stem cells， and drive the regeneration after injury.

Study on three dimensional printing of fetal aortic arch and its branches anomalies / 中华超声影像学杂志

Objective:To explore the feasibility and application value of three dimensional (3D) printing technology in creating models of abnormal fetal aortic arch and its branches.Methods:Eleven cases of abnormal fetuses confirmed fetal aortic arch and its branches anomalies from March 2019 to July 2020 in Renmin Hospital of Wuhan University were prospectively enrolled. All the fetuses underwent two dimensional(2D) echocardiography and spatio-temporal image correlation (STIC) technology examination. The 3D volume images of fetal heart were post-processed by Mimics software to create images of the great vessels and their branches in standard tessellation language format (STL) file. The STL file was output to the 3D printer and the 3D printing models of fetal great vessels and their branches were obtained. Compared with conventional ultrasound, the characteristics and application value of 3D printed models of abnormal fetal aortic arch and its branches were analyzed.Results:Eleven fetuses were successfully modeled and printed out large blood vessels and their branch models. The 3D printing model had its own advantages in displaying large blood vessels and their branch abnormalities. It could provide high quality imaging anatomical details and visualize great vessels origin, branch and position and can better display vascular ring spatial relations.Conclusions:It is feasible to use 3D printing technology to make the fetal aortic arch and its branch abnormal model. The 3D printing model can directly display its characteristic changes and provide a certain reference basis for accurately determining the type of vascular ring in the prenatal stage.

Chemical Composition Analysis of Platycladi Cacumen Before and After Being Carbonized Based on Identification by UHPLC-QTOF-MS/MS / 中国实验方剂学杂志

Objective:To identify the chemical constituents of Platycladi Cacumen<italic> </italic>before and after being carbonized. Method:Chemical constituents in 3 batches of Platycladi Cacumen and its carbonized products<italic> </italic>were identified and compared by ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-QTOF-MS/MS）. Chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with 0.1% formic acid aqueous solution （A）- acetonitrile （B） as mobile phase for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B； 18-22 min， 100%B）. The flow rate was 0.4 mL·min^-1 and the injection volume was 5 μL. Mass spectrometry was performed by an electrospray ionization， and the primary and secondary mass spectrometry data were collected with the full scan mode of positive and negative ions， the peaks containing MS/MS data were identified by self-established secondary mass spectrometry database and corresponding fragmentation law matching method. Result:A total of 77 and 76 substances with the same change trend were identified under positive and negative ion modes. After being<italic> </italic>carbonized， the disappeared components of Platycladi Cacumen were mainly amino acids， ketone aldehydes and other volatile components. Among newly produced components， there were 6 kinds of flavonoid aglycones （rhamnetin， 6，7，3'-trihydroxyflavone， 3，6，3'-trihydroxyflavone， 4'-hydroxy-2'-methyl-3，4，5-trimethoxychalcone， herbacetin and 3'，5'-dimethoxy-3，5，7，4'-tetrahydroxyflavone）， 3 kinds of coumarins （7-hydroxycoumarin， 7，8-dihydroxycoumarin and 8-acetyl-7-hydroxycoum-arin） and 3 kinds of benzoic acids （3-methylcatechol， pyrocatechol and chromone-3-carboxylic acid）. There were a total of 40 flavonoids （quercitrin， quercetin， kaempferol， etc.） among these identified chemical constituents. Conclusion:There are significant quantitative and qualitative changes in the chemical compositions of Platycladi Cacumen after being carbonized. The flavonoids， the identified main active ingredients， can provide data reference for further study on the material basis of efficacy changes of Platycladi Cacumen<italic> </italic>before and after being carbonized.

Semen parameters in men recovered from COVID-19 / 亚洲男科学杂志(英文版)

The novel coronavirus disease (COVID-19) pandemic is emerging as a global health threat and shows a higher risk for men than women. Thus far, the studies on andrological consequences of COVID-19 are limited. To ascertain the consequences of COVID-19 on sperm parameters after recovery, we recruited 41 reproductive-aged male patients who had recovered from COVID-19, and analyzed their semen parameters and serum sex hormones at a median time of 56 days after hospital discharge. For longitudinal analysis, a second sampling was obtained from 22 of the 41 patients after a median time interval of 29 days from first sampling. Compared with controls who had not suffered from COVID-19, the total sperm count, sperm concentration, and percentages of motile and progressively motile spermatozoa in the patients were significantly lower at first sampling, while sperm vitality and morphology were not affected. The total sperm count, sperm concentration, and number of motile spermatozoa per ejaculate were significantly increased and the percentage of morphologically abnormal sperm was reduced at the second sampling compared with those at first in the 22 patients examined. Though there were higher prolactin and lower progesterone levels in patients at first sampling than those in controls, no significant alterations were detected for any sex hormones examined over time following COVID-19 recovery in the 22 patients. Although it should be interpreted carefully, these findings indicate an adverse but potentially reversible consequence of COVID-19 on sperm quality.

Health self-assessment of the migrant workers received physical examination in Shanghai / 上海预防医学

Objective:To understand the health self-assessment and related influencing factors in the migrant workers who received of physical examination in Shanghai， so as to provide reference for formulating health service policies for migrant workers. Methods:Taking Shanghai "Intelligent Blue Collar" Health Science Popularization Service Station as the investigation site， 8，100 urban workers were surveyed by questionnaire. The questionnaire includes basic information， health self-assessment， basic medical insurance coverage and satisfaction， subjective well-being and so on. Results:There were significant differences between migrant workers and local workers in health self-assessment， two-week disease prevalence rate， and chronic disease prevalence rate（χ2=32.981，P<0.05；χ2=4.554，P<0.05；χ2=86.695，P<0.05）. The differences between migrant workers and local workers were also significant in the basic medical insurance coverage rate and the level of satisfaction （χ2=221.592，P<0.05；χ2=30.950，P<0.05）. There was no significant difference in subjective well-being between these two groups（χ2=8.787，P>0.05）. The health self-assessment of older migrant workers was significantly better than that of new generation migrant workers（β=0.062，P<0.05）， and was significantly worse in female migrant workers than that in male migrant workers（β=-0.120，P<0.05）. The health self-assessment was significantly better in migrant workers with agricultural household registration than that in non-agricultural household registration（β=0.060，P<0.05）. The higher the satisfaction of basic medical insurance，the higher the subjective well-being and the better the health self-assessment（β=0.148，P<0.05；β=0.422，P<0.05）. Conclusion:The health status of migrant workers in Shanghai who received physical examination is better than that of local workers， and the participation rate of basic medical insurance is lower than that of local workers. Age， gender and household registration are the important factors influencing the health self-assessment. Satisfaction with basic medical insurance level and subjective well-being is a significant factor to predict health self-assessment positively . We should further improve the fairness of medical insurance benefits for migrant workers， improve subjective well-being， and ensure the health fairness of the group.

Novel biallelic loss-of-function mutations in / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a specific type of asthenoteratozoospermia, presenting with multiple morphological anomalies in spermatozoa, such as absent, bent, coiled, short, or irregular caliber flagella. Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins (CFAPs; e.g., CFAP43, CFAP44, CFAP65, CFAP69, CFAP70, and CFAP251) responsible for the MMAF phenotype in infertile men from different ethnic groups. However, none of them have been identified in infertile Pakistani males with MMAF. In the current study, two Pakistani families with MMAF patients were recruited. Whole-exome sequencing (WES) of patients and their parents was performed. WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families (Family 1: ENST00000357060.3, p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state; Family 2: ENST00000357060.3, p.Thr526Serfs*43 in a homozygous state). Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype. Semiquantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out to detect the effect of the mutation on mRNA of the affected gene. Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients. To the best of our knowledge, this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype. This study will help researchers and clinicians to understand the genetic etiology of MMAF better.

The molecular control of meiotic double-strand break (DSB) formation and its significance in human infertility / 亚洲男科学杂志(英文版)

Meiosis is an essential step in gametogenesis which is the key process in sexually reproducing organisms as meiotic aberrations may result in infertility. In meiosis, programmed DNA double-strand break (DSB) formation is one of the fundamental processes that are essential for maintaining homolog interactions and correcting segregation of chromosomes. Although the number and distribution of meiotic DSBs are tightly regulated, still abnormalities in DSB formation are known to cause meiotic arrest and infertility. This review is a detailed account of molecular bases of meiotic DSB formation, its evolutionary conservation, and variations in different species. We further reviewed the mutations of DSB formation genes in association with human infertility and also proposed the future directions and strategies about the study of meiotic DSB formation.

Efficacy and Safety of Endoscopic Treatment for Gastrointestinal Stromal Tumors in the Upper Gastrointestinal Tract

Background/Aims@#Endoscopic treatment (ET) has been applied for decades to treat subepithelial tumors, including gastrointestinal stromal tumors (GISTs). However, the efficacy of ET remains debatable. In this study, we evaluated the efficacy and safety of ET for GISTs in the upper gastrointestinal tract. @*Methods@#This retrospective single-center study included 97 patients who underwent ET. All patients were enrolled from July 2014 to July 2018. Parameters such as demographics, size, resection margin, complications, pathological features, procedure time, total cost, and follow-up were investigated and analyzed. @*Results@#Our study achieved 100% en bloc resection and 77.4% (72/93) R0 resection. The most common location was the fundus with a mean tumor size of 2.1±1.43 cm. The mean age, procedure time, hospital stay, and cost were 59.7±11.29 years, 64.7±35.23 minutes, 6.8 days, and 5,337 dollars, respectively. According to National Institutes of Health classification, 63 (64.9%), 26 (26.8%), 5 (5.2%), and 3 (3.1%) patients belonged to the very low, low, intermediate, and high risk classification, respectively. Immunohistochemistry results showed a 100% positive rate of CD34, DOG-1, CD117, and Ki67. A mean follow-up of 21.3±13.0 months showed no recurrence or metastasis. @*Conclusions@#ET is effective and safe for curative removal of GISTs in the upper gastrointestinal tract, and it can be a treatment of choice for patients with no metastasis.

Primary study on scientific connotation "carbonizing retains characteristics" of Platycladi Cacumen / 中草药

Objective: To study the scientific connotation "carbonizing retains characteristics" of Platycladi Cacumen (PC). Methods: Chemical constituents of PC and Platycladi Cacumen Carbonisata (PCC) of different carbonizing degree were compared by HPLC characteristic chromatogram, and the chromatographic peaks were assigned; PC and PCC of different carbonizing degree and the characteristic components changed before and after being carbonized were used to compare the impact to hemostasis in vitro and inhibition of zebrafish cerebral hemorrhage. Results: Chemical compositions of PC were affected by different carbonizing degree, when carbonizing degree is moderate, the content of myricitrin, quercitroside, isoquercitroside, amentoflavone and hinokiflavone original in PC were significantly reduced and the amount of quercetin and kaempferol which were newly produced were higher. In vitro hemostatic experiments showed that compared with the blank group, the APTT and FIB of PC which was carbonized moderately were significantly different (P < 0.01) and TT was significantly different (P < 0.05). It also had obvious inhibition effect on zebrafish cerebral hemorrhage when the concentration of PC carbonized moderately was 50 μg/mL (P < 0.01). The enhanced hemostasis was significantly related to newly produced compositions quercetin and kaempferol. Conclusion: Chemical compositions of PC changed significantly after being carbonized and the hemostatic effect was enhanced,which were related to the processing degree. The scientific connotation and traditional processing requirements of "carbonizing retains characteristics" was preliminarily analyzed when the carbon medicine was processed by this study, which provides a certain idea for the research on traditional processing theory of carbon medicine.

Analysis of volatile organic compounds (VOCs) fingerprint of raw and honey-fried licorice based on headspace-gas-chromatography ion-mobility spectrometry (HS-GC-IMS) / 中国中药杂志

Licorice is one of the most commonly used traditional Chinese medicine. In clinic, raw licorice and honey-fried licorice are used in medicines, with the main effects in clearing away heat and detoxifying, moistening lungs and removing phlegm. Honey-fried licorice has effects in nourishing the spleen and stomach and replenishing Qi and pulse. Because traditional Chinese medicine exerts the effects through multiple components and multiple targets, the index components used in the quality evaluation of licorice are often difficult to reflect their real quality. In addition, most of studies for the quality standards have shown that honey-fried licorice are the same as licorice, with a lack of quality evaluation standards that can demonstrate their processing characteristics. The quality of medicine is directly related to its clinical efficacy, so it is necessary to establish a more effective quality control method. Licorice has a beany smell, which is one of the main quality identification characteristics. In this study, by taking advantage of the odor characteristics, a headspace-gas chromatography-ion migration mass spectrometry technology was used to establish a quality evaluation method. A total of 76 volatile components were identified. Through the dynamic principal component analysis, 7 kinds of volatile substances in raw licorice and 13 kinds of volatile substances in honey-fried licorice were statistically obtained, and could be taken as index components for the quality evaluation of raw and honey-fried licorice, respectively. This study could help realize the combination and unification of modern detection and traditional quality evaluation methods, and make a more realistic evaluation for the quality of licorice.

Gut microbiota dysbiosis in preeclampsia patients in the second and third trimesters / 中华医学杂志(英文版)

BACKGROUND@#Preeclampsia (PE) is a serious complication that affects maternal and perinatal outcomes. However, the mechanisms have not been fully explained. This study was designed to analyze longitudinal gut microbiota alterations in pregnant women with and without PE in the second (T2) and third trimesters (T3).@*METHODS@#In this nested case-control study, which was conducted at Nanjing Maternity and Child Health Care Hospital, fecal samples from 25 PE patients (25 fecal samples obtained in T2 and 15 fecal samples obtained in T3) and 25 matched healthy controls (25 fecal samples obtained in T2 and 22 fecal samples obtained in T3) were collected, and the microbiota were analyzed using 16S rRNA gene sequencing. The diversity and composition of the microbiota of PE cases and controls were compared.@*RESULTS@#No significant differences in diversity were found between the PE and control groups (P > 0.05). In the control group, from T2 to T3, the relative abundances of Proteobacteria (median [Q1, Q3]: 2.25% [1.24%, 3.30%] vs. 0.64% [0.20%, 1.20%], Z = -3.880, P < 0.05), and Tenericutes (median [Q1, Q3]: 0.12% [0.03%, 3.10%] vs. 0.03% [0.02%, 0.17%], Z = -2.369, P < 0.05) decreased significantly. In the PE group, the relative abundance of Bacteroidetes in T2 was lower than in T3 (median [Q1, Q3]: 18.16% [12.99%, 30.46%] vs. 31.09% [19.89%, 46.06%], Z = -2.417, P < 0.05). In T2, the relative abundances of mircrobiota showed no significant differences between the PE group and the control group. However, in T3, the relative abundance of Firmicutes was significantly lower in the PE group than in the control group (mean ± standard deviation: 60.62% ± 15.17% vs. 75.57% ± 11.53%, t = -3.405, P < 0.05). The relative abundances of Bacteroidetes, Proteobacteria, and Enterobacteriaceae were significantly higher in the PE group than in the control group (median [Q1, Q3]: 31.09% [19.89%, 46.06%] vs. 18.24% [12.90%, 32.04%], Z = -2.537, P < 0.05; 1.52% [1.05%, 2.61%] vs. 0.64% [0.20%, 1.20%], Z = -3.310, P < 0.05; 0.75% [0.20%, 1.00%] vs. 0.01% [0.004%, 0.023%], Z = -4.152, P < 0.05). Linear discriminant analysis combined effect size measurements analysis showed that the relative abundances of the phylum Bacteroidetes, class Bacteroidia and order Bacteroidales were increased in the PE group, while those of the phylum Firmicutes, the class Clostridia, the order Clostridiales, and the genus unidentified Lachnospiraceae were decreased in the PE group; and these differences were identified as taxonomic biomarkers of PE in T3.@*CONCLUSION@#From T2 to T3, there was an obvious alteration in the gut microbiota. The gut microbiota of PE patients in T3 was significantly different from that of the control group.