Microbial communities such as those residing in the human gut are highly diverse and complex, and many with important implications for health and diseases. The effects and functions of these microbial communities are determined not only by their species compositions and diversities but also by the dynamic intra- and inter-cellular states at the transcriptional level. Powerful and scalable technologies capable of acquiring single-microbe-resolution RNA sequencing information in order to achieve a comprehensive understanding of complex microbial communities together with their hosts are therefore utterly needed. Here we report the development and utilization of a droplet-based smRNA-seq (single-microbe RNA sequencing) method capable of identifying large species varieties in human samples, which we name smRandom-seq2. Together with a triple-module computational pipeline designed for the bacteria and bacteriophage sequencing data by smRandom-seq2 in four human gut samples, we established a single-cell level bacterial transcriptional landscape of human gut microbiome, which included 29,742 single microbes and 329 unique species. Distinct adaptive response states among species in Prevotella and Roseburia genera and intrinsic adaptive strategy heterogeneity in Phascolarctobacterium succinatutens were uncovered. Additionally, we identified hundreds of novel host-phage transcriptional activity associations in the human gut microbiome. Our results indicated that smRandom-seq2 is a high-throughput and high-resolution smRNA-seq technique that is highly adaptable to complex microbial communities in real-world situations and promises new perspectives in the understanding of human microbiomes.
Humans ; Gastrointestinal Microbiome/genetics* ; Bacteriophages/physiology* ; High-Throughput Nucleotide Sequencing ; Sequence Analysis, RNA/methods* ; Bacteria/virology*

Objective:To analyze the etiological characteristics of epidemic cerebrospinal meningitis caused by serogroup Y Neisseria meningitidis ( Nm) in Fujian Province. Methods:The strain identification, serogroup identification, antimicrobial susceptibility test, molecular typing, average nucleotide identity (ANI), core genome multilocus sequence typing (cgMLST), and single nucleotide polymorphism (SNP) analysis were performed on eight Nm strains isolated from cases of epidemic cerebrospinal meningitis, close contacts, or healthy population. Results:All eight isolates from Fujian Province in 2024 were serogroup Y Nm strains. The antimicrobial susceptibility test for 12 antibiotics showed that all isolates from Fujian Province in 2024 were resistant to trimethoprim-sulfamethoxazole, and NMFJ202406 and NMFJ202407 isolates were resistant to nalidixic acid and intermediate to ciprofloxacin, while the others were sensitive to antibiotics such as penicillin, cefotaxime, and ceftriaxone. All isolates were ST-1655 sequence type (ST), except for NMFJ202406 isolate which were ST-2039, and all isolates belonged to clonal complex 23 (CC23). The finetyping antigen profile of all isolates were P1.5-1, 10-1: F4-1. ANI and phylogenetic analysis indicated that the isolates in Fujian Province were mainly divided into two clusters, namely NMFJ202401 and its close contact isolates (NMFJ202402-NMFJ202405), as well as NMFJ202406, NMFJ202407, and NMFJ202408 isolates. Compared to the isolates from Guangdong, Shanghai, and Hebei Provinces, the strains isolated in Fujian Province were more closely related to the strains isolated in Canada (NML2019-167, 2019), Japan (NIID501, 2009), and the United Kingdom (CA41956, 2015), respectively. Conclusions:This is the first report of epidemic cerebrospinal meningitis caused by serogroup Y Nm isolates in Fujian Province, which might be transmitted from different sources abroad and have spread in many areas. In the future, the wider usage of tetravalent meningococcal vaccine ACYW135 is needed, and the epidemiological and etiological surveillance of serogroup Y epidemic cerebrospinal meningitis should be strengthened.

Gastric cancer is one of the malignancies with high incidence in the world. Xiangsha Liu Junzitang，a common prescription for the prevention and treatment of gastric cancer，has the effects of moving Qi to relieve pain，drying dampness， and invigorating the spleen. It is especially indicated for gastric cancer of the spleen and stomach qi deficiency syndrome. Based on the databases such as CNKI，Wanfang Data，and PubMed，the clinical efficacy and experimental studies of Xiangsha Liu Junzitang for the prevention and treatment of gastric cancer were summarized and sorted out，and the mechanism of Xiangsha Liu Junzitang for the prevention and treatment of gastric cancer was elaborated in order to provide useful references for the clinical and basic research on Xiangsha Liu Junzitang in the field of gastric cancer in the future. In clinical practice，Xiangsha Liu Junzitang can treat gastric precancerous lesions，increase the body immunity of patients with gastric cancer，improve the symptoms of spleen and stomach weakness after gastric cancer surgery，and reduce the adverse reactions of the digestive tract after chemotherapy for gastric cancer. Its clinical efficacy is superior to that of western medicine alone whether it is combined with western medicine or used alone. In the experimental research，Xiangsha Liu Junzitang has the effects of regulating inflammatory factors，inhibiting the proliferation of gastric cancer cells，promoting the apoptosis of gastric cancer cells，and improving the activity of pepsin. Modern pharmacological research has shown that Xiangsha Liu Junzitang can conduct a comprehensive intervention with multiple components and multiple targets. The main components of a single drug contained include saponins，polysaccharides，lactones，volatile oils，organic acids，and others， with the effects of protecting gastric mucosa，regulating endocrine，and promoting apoptosis of epithelial cells in gastric mucosal dysplasia，reflecting the advantages and values of traditional Chinese medicine in the prevention and treatment of gastric cancer.

Objective:To assess the efficacy and safety of 100 units of botulinum toxin A (BTX-A) intradetrusor injection in patients with overactive bladder.Methods:From April 2016 to December 2018, 17 tertiary hospitals were selected to participate in this prospective, multicenter, randomized, double-blind, placebo-controlled study. Two phases of study were conducted: the primary phase and the extended phase. This study enrolled patients aged 18 to 75 years who had been inadequately managed by anticholinergic therapy (insufficient efficacy or intolerable side effects) and had spontaneous voiding with overactive bladder. Exclusion criteria included patients with severe cardiac, renal and hepatic disorders, patients with previous botulinum toxin treatment for 6 months or allergic to BTX-A, patients with urinary tract infections, patients with urinary stones, urinary tract tumors, diabetes mellitus, and bleeding tendency. Eligible patients were randomly assigned to BTX-A group and placebo control group in a ratio of 2∶1. Two groups of patients received 20 intradetrusor injections of BTX-A 100U or placebo at the depth of the submucosal muscle layer respectively under cystoscope, including 5 injections at the base of the bladder, 3 injections to the bladder triangle, 5 injections each to the left and right walls and 2 injections to the top, sparing the bladder neck. As a placebo control group, patients received same volume of placebo containing no BTX-A and only adjuvant freeze-dried preparations for injection with the same method. A combination of gelatin, sucrose, and dextran served as adjuvants. Average micturition times per 24 hours, urinary incontinence (UI) episodes per day, average micturition volume per day, OAB symptom score(OABSS), and quality of life (QOL) score were recorded at baseline and the 2nd, 6th and 12th week after treatment. The primary efficacy endpoint was the change from baseline in the average micturition times per 24 hours at the 6th week after treatment. The secondary efficacy endpoints included the change from baseline in the average micturition times per 24 hours at 2nd and 12th week, as well as the change from baseline in the OABSS, QOL score, average frequency of urgency and UI episodes per day, urgency score, average micturition volume per day at 2nd, 6th and 12th week after treatment. Patients were followed for 12 weeks to assess adverse events (AEs). After assessed at week 12, if the micturition times has decreased less than 50% compared to baseline and the patient is willing to receive retreatment, then patients could enter the extended trial phase. In that phase, patients in both groups were injected with 100 units BTX-A from 12th week onwards and then followed up the same indicators for 12 weeks.Results:216 patients were enrolled in this trial (144 cases in the BTX-A group and 72 cases in the placebo control group). Baseline characteristics such as age (47.75±14.20 in the BTX-A group and 46.39±15.55 in the control group), sex (25 male/117 female in the BTX-A group and 10/61 in the control group), and disease duration (0.51 years in the BTX-A group and 0.60 years in the control group) were balanced between the two groups( P>0.05). A marked reduction from baseline in average micturition times per 24 hours was observed in all treatment groups at the 6th week and the reduction of the two groups was statistically different ( P<0.001 and P=0.008 respectively). Compared with the baseline, the average micturition times per 24 hours at the 6th week decreased from baseline by 2.40(0.70, 4.60)times for the BTX-A group and 0.70(-1.00, 3.30) times for the placebo control group respectively, and the difference between the two groups was considered to be statistically significant ( P=0.003). The change rates of average micturition times per 24 hours from baseline at the 6th week of the two groups were (16±22)% and (8±25)% respectively, and the difference between the two groups was statistically significant ( P=0.014). Compared with the baseline, the average micturition times per 24 hours at 2nd and 12th week decreased by 2.00(0.00, 4.00)and 3.30(0.60, 5.03)for the BTX-A group, 1.00(-1.00, 3.00)and 1.70(-1.45, 3.85)for the placebo control group respectively. The difference between two groups was considered to be statistically significant ( P=0.038 and P=0.012); the changes of average urgency times per day for the BTX-A group and the control group at the 2nd, 6th and 12th week were 2.00(0.00, 4.30)and 2.40(0.30, 5.00), 3.00(0.30, 5.70)and 0.70(-1.30, 2.70), 0.70(-1.30, 3.00) and 1.35(-1.15, 3.50), respectively. There were significant differences between two groups at the 2nd, 6th and 12th week, ( P=0.010, P=0.003 and P=0.025, respectively). The OABSS of the BTX-A group and the control group at the 6th week decreased by 1.00(0.00, 4.00)and 0.50(-1.00, 2.00) compared with the baseline, and the difference between the two groups was statistically significant ( P=0.003). 47 cases of BTX-A group and 34 cases of placebo control group entered the extended trial phase, and 40 and 28 cases completed the extended trial phase, respectively. The average micturition volume per 24 hours changed by -16.60(-41.60, -0.60)ml and -6.40(-22.40, 13.30)ml, (-35.67±54.41)ml and(-1.76±48.69)ml, (-36.14±41.51)ml and (-9.28±44.59)ml, (-35.85±43.35)ml and(-10.41±40.29)ml for two groups at the 12th, 14th, 18th and 24th week, and the difference between two groups was statistically significant at each follow-up time ( P=0.01, 0.006, 0.012 and 0.016, respectively). There was no significant difference in other parameters( P>0.05). However, adverse reactions after intradetrusor injection included increased residual urine volume (27 in the BTX-A group and 3 in the control group), dysuria (21 in the BTX-A group and 6 in the control group), urinary infection (19 in the BTX-A group and 6 in the control group), bladder neck obstruction (3 in the BTX-A group and 0 in the control group), hematuria (3 in the BTX-A group and 1 in the control group), elevated alanine aminotransferase (3 in the BTX-A group and 0 in the control group), etc. During the follow-up period, there was no significant difference in the other adverse events between two groups except the increase of residual urine volume( P<0.05). In the primary trial phase, among the 27 cases with increased residual urine volume in BTA group, only 1 case (3.70%) with PVR more than 300 ml; the PVR of 3 patients in the placebo group was less than 100 ml. The increase of residual urine volume caused by the injection could be improved or disappeared with the passage of time. Conclusions:Intradetrusor injection of Chinese BTX-A improved the average micturition times per 24 hours, the average daily urgent micturition times, OABSS, and average micturition volume per time, and reduced the adverse effects in patients with overactive bladder.Chinese BTX-A at dose of 100U demonstrated durable efficacy and safety in the management of overactive bladder.

[Objective] To analyze the clinical curative effect of 20G and 23G vitrectomy for posterior segment intraocular foreign bodies,to explore the differences of their efficacy and safety.[Methods] This was a retrospective case study.Select 71 patients (71 eyes) who suffered from posterior segment intraocular foreign bodies and underwent different ways of vitrectomy,according to the way of vitrectomy,the patients were enrolled into 20G vitrectomy group (20G group,37 patients,37 eyes) and 23G vitrectomy group (23G group,34 patients,34 eyes).All patients were given wound suture,and patients complicated traumatic cataract should underwent cataract surgery;then were given 20G/23G vitrectomy and extraction of intraction foreign bodies,and (or) be given retinal laser photocoagulation,cryocoagulation,and endotamponade during the procedure.Extraction of intraction foreign bodies and the location of retinal,surgical time,postoperative inflammation and stimulus syndrome,length of hospital stay,the best corrected acuity (BCVA) and other complications after surgery were registered.Minimum follow-up was 6 months.[Results] The rate of extraction of intraction foreign bodies and early retinal reattachment rate were 100％ of the two groups.Comparing the complications after surgery between the two groups,the differences had no statistical significance (all P ＞ 0.05) except the incidence of postoperative ocular hypotenison.The significant difference was found in the comparison of surgical time,average hospitalizcd days,postoperative inflammation score and stimulus syndrome between the two groups,and there were significant differences when the vision distribution before and after surgery in two groups were self-compared (P ＜ 0.05).At lastest follow up,the differences had no statistical significance when comparing the cases of unplanned surgical reoperation,the location of retinal and the vision distribution between the two groups (P ＞ 0.05).[Conclusion] 20G and 23G vitrectomy are both safely and effectively performed in patients with posterior segment intraocular foreign bodies,the efficacy and safety of them are comparable.There is higher incidence of postoperative ocular hypotension in 23G group,and it has an ascendant than 20G group in shorter surgical time and length of hospital stay,lighter postoperative inflamnation and stimulus syndrome.

We investigated the epidemiological characteristics of diarrheagenic Escherichia coli (E.coli) infection in Fujian Province,providing reference for the prevention and control of diarrhea epidemic and outbreak.Yanping and Yongan areas were selected as surveillance sites from year 2010 to 2015,where 1 950 samples were collected from the sentinel hospitals,and then samples were isolated and cultured on MacConkey agar plates.Suspected strains were identified by routine and molecular diag nosis technique methods.Results were analyzed by SPSS 17.0 software package.A total of 129 strains of diarrheagenic E.coli were isolated with a total detection rate of 6.62％.The detection rates of enteropathogenic E.coli(EPEC),enterotoxigenic E.coli(ETEC) and enteroadhesive E.coli(EAEC) were 3.33％,1.64％ and 1.64％ respectively (including 61 strains of atypical EPEC,4 strains of typical EPEC,and 32 strains of ETEC and EAEC for each),while EHEC and EIEC were not found.There was no significant difference between the detection rates of male and female.All the patients were divided into five groups according to the age,and there were no significant differences between the detection rates of EPEC,EAEC and ETEC in the 5 groups.The detection rates were the highest in August and September.There was no significant difference between the detection rates of rural area and uban area.There was also no significant difference between the composition ratio of diarrheagenic E.coli (DEC) in the two surveillance sites.In conclusion,there were three kinds of DEC in Fujian Province,and EPEC was the dominant.August and September were the months with the highest detection rates.Children age less than five and adult aged 20 59 years were the high risk groups with DEC infection.More attention should be attached on the rational treatment by antibiotic for DEC.

The emergence of atypical El Tor strains from V .cholerae in South Asia and Africa has been attributed to several outbreaks in recent decades ,however ,backgrounds of such strains in China remain exclusive .In this study ,PCR am-plification of both El Tor and classical alleles for ctxB ,tcpA ,rstR and hlyA genes was attempted in sixty-nine El Tor isolates from Fujian between 1962 to 2005 ,in addition ,some amplicons were sequence-analyzed .Thus ,the time point of atypical EVC strains in Fujian was determined ,genetic diversities of such strains were investigated .It was revealed that ctxB-Cl ,tcpA-Cl and hlyA genes were detected in O1 serogroup EVC isolates from Fujian since 1962 .Although rstR-Cl gene was solely detected in isolates between 1994 to 2000 .It was indicated by sequence analysis that atypical EVC strains from Fujian possessed a novel T→G mutation at residual 204 of the ctxB gene .Remarkably ,two novel ctxB genotypes (ctxB-10 and ctxB-11) were identified in one strain .The residual 115-C of ctxB in ctxB-11 showed characteristics of ctxB-Cl ,however ,its residual 203-T demonstra-ted characteristics of ctxB-El .This observation implied that it was common in O1 serogroup EVC strains from Fujian hybrid-ized with classical alleles since 1962 ,which would be the earliest time-point for the emergence of atypical El Tor strains hitherto in literature .Emergence of atypical El Tor strains harboring rstR-Cl in Fujian occurred since 1994 .Meanwhile ,novel mutation sites and ctxB genotypes were observed in Fujian isolates ,including diverse combination of ctxB genotypes in one strain and combination of biotype-specific sites in ctxB sequences .In summary ,molecular characterization of O 1 serogroup EVC strains from Fujian was unique and geography-associated .