First Report of Root-knot Nematode, Meloidogyne enterolobii on Passiflora edulis in Yulin, China.

Passion fruit (Passiflora edulis), a medicinal plant, was introduced into China in the early 19th century, is mainly cultivated in southern provinces (Liang et al. 2019). During March 2023, a survey was carried out and 167 samples were taken from passion fruit cultivated area in Yulin (22.6570263°E; 110.1765019°N) apart from the planting base appeared yellow leaves, stunted growth, and distinctive galls on the roots. Within the galls, Meloidogyne sp. females and egg masses were observed. From the rhizosphere soil, second-stage juveniles (J2) were extracted, and population density was 105/500 g soil. The species was determined to be Meloidogyne enterolobii based on morphological characteristics, including female perineal pattern, and genetic analyses. Female (n = 10) perineal patterns showed oval shape, with coarse and smooth striae, dorsal arch rounded to square, and lateral lines not distinct. The male head cap was high and rounded, with the head region only slightly set off from the body, knobs large, ovoid to rounded. The measurements of males (n = 10) included body length, 1,230.7 ± 244.94 (997 to 1,569) µm; a, 38.58 ± 7.8 (33.45 to 47.05) µm; c, 113.03 ± 26.22 (80.82 to 144.23) µm; stylet, 15.68 ± 1.1 (14.5 to 17.4) µm; spicules, 31.83 ± 2.84 (28.69 to 36.1) µm; tail, 11.09 ± 1.72 (8.02 to 13.38) µm; and gubernaculum length, 8.34 ± 0.28 (8.11 to 8.98) µm. Measurements of J2 (n = 20) included body length, 455.75 ± 44.94 (381 to 512) µm; a, 26.32 ± 3.89 (18.18 to 32.70) µm; c, 8.56 ± 1.2 (6.36 to 10.80) µm; stylet, 12.44 ± 0.76 (11.2 to 13.8) µm; DGO, 3.65 ± 0.54 (2.84 to 4.68) µm; tail, 53.89 ± 6.36 (39.8 to 62.2) µm; and hyaline tail terminus, 11.77 ± 2.83 (7.14 to 16.2) µm. These morphological characteristics are similar to those reported in the original description of M. enterolobii (Yang and Eisenback 1983). The sequences of the partial ITS region was amplified with V5367 (5'-TTGATTACGTCCCTGCCCTTT-3') and 26S (5'-TTTCACTCGCCGTTACTAAGG-3') primers (Vrain et al. 1992). The region between cytochrome oxidase subunit II (COII) and the 16S rRNA mitochondrial DNA (mtDNA COII) was also amplified with the primers C2F3 (5'-GGTCAATGTTCAGAAATTTGTGG-3') (Powers and Harris 1993) and MRH106 (5'-AATTTCTAAAGACTTTTCTTAGT-3') (Stanton et al. 1997). The ITS region yielded a fragment of 757 bp (OR072957) and mtDNA COII of 706 bp (OR078415). A BLAST search indicated the sequences were 100% identical to several sequences of M. enterolobii (MT406250, MH756127 and AY831967, MN269940, respectively). To confirm pathogenicity, 20 passion fruit (P. edulis Sim. f. flavicarpa) 30-day-old seedlings were transplanted into pots with an autoclaved mixture of sand and field soil (3:1) and maintained in the glasshouse at 25 ± 2°C with 65 ± 5% relative humidity. After eight weeks, fifteen plants were inoculated with 500 J2/pot (nematode culture collected from the original field), and another five uninoculated plants served as a control. Two months later, aboveground symptoms were similar to those observed in the field. Nematode reproduction occurred and root galls were observed. The reproduction factor (nematode final population density/initial population density) was 4.8. The disease caused by M. enterolobii was severe in Yulin city of Guangxi. Guangxi is an important area for passion fruit culture, with about 2000 ha, which is responsible for two-thirds of China production (Xing et al. 2020). This is the first record of P. edulis natural infection with M. enterolobii in the Yulin City of Guangxi, China.

[Safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy: A prospective, multi-center, single arm trial].

Objective: To evaluate the safety of double and a half layered esophagojejunal anastomosis in radical gastrectomy. Methods: This prospective, multi-center, single-arm study was initiated by the Affiliated Cancer Hospital of Zhengzhou University in June 2021 (CRAFT Study, NCT05282563). Participating institutions included Nanyang Central Hospital, Zhumadian Central Hospital, Luoyang Central Hospital, First Affiliated Hospital of Henan Polytechnic University, First Affiliated Hospital of Henan University, Luohe Central Hospital, the People's Hospital of Hebi, First People's Hospital of Shangqiu, Anyang Tumor Hospital, First People's Hospital of Pingdingshan, and Zhengzhou Central Hospital Affiliated to Zhengzhou University. Inclusion criteria were as follows: (1) gastric adenocarcinoma confirmed by preoperative gastroscopy;(2) preoperative imaging assessment indicated that R0 resection was feasible; (3) preoperative assessment showed no contraindications to surgery;(4) esophagojejunostomy planned during the procedure; (5) patients volunteered to participate in this study and gave their written informed consent; (6) ECOG score 0-1; and (7) ASA score I-III. Exclusion criteria were as follows: (1) history of upper abdominal surgery (except laparoscopic cholecystectomy);(2) history of gastric surgery (except endoscopic submucosal dissection and endoscopic mucosal resection); (3) pregnancy or lactation;(4) emergency surgery for gastric cancer-related complications (perforation, hemorrhage, obstruction); (5) other malignant tumors within 5 years or coexisting malignant tumors;(6) arterial embolism within 6 months, such as angina pectoris, myocardial infarction, and cerebrovascular accident; and (7) comorbidities or mental health abnormalities that could affect patients' participation in the study. Patients were eliminated from the study if: (1) radical gastrectomy could not be completed; (2) end-to-side esophagojejunal anastomosis was not performed during the procedure; or (3) esophagojejunal anastomosis reinforcement was not possible. Double and a half layered esophagojejunal anastomosis was performed as follows: (1) Open surgery: the full thickness of the anastomosis is continuously sutured, followed by embedding the seromuscular layer with barbed or 3-0 absorbable sutures. The anastomosis is sutured with an average of six to eight stitches. (2) Laparoscopic surgery: the anastomosis is strengthened by counterclockwise full-layer sutures. Once the anastomosis has been sutured to the right posterior aspect of the anastomosis, the jejunum stump is pulled to the right and the anastomosis turned over to continue to complete reinforcement of the posterior wall. The suture interval is approximately 5 mm. After completing the full-thickness suture, the anastomosis is embedded in the seromuscular layer. Relevant data of patients who had undergone radical gastrectomy in the above 12 centers from June 2021 were collected and analyzed. The primary outcome was safety (e.g., postoperative complications, and treatment). Other studied variables included details of surgery (e.g., surgery time, intraoperative bleeding), postoperative recovery (postoperative time to passing flatus and oral intake, length of hospital stay), and follow-up conditions (quality of life as assessed by Visick scores). Result: [1] From June 2021 to September 2022,457 patients were enrolled, including 355 men and 102 women of median age 60.8±10.1 years and BMI 23.7±3.2 kg/m2. The tumors were located in the upper stomach in 294 patients, mid stomach in 139; and lower stomach in 24. The surgical procedures comprised 48 proximal gastrectomies and 409 total gastrectomies. Neoadjuvant chemotherapy was administered to 85 patients. Other organs were resected in 85 patients. The maximum tumor diameter was 4.3±2.2 cm, number of excised lymph nodes 28.3±15.2, and number of positive lymph nodes five (range one to four. As to pathological stage,83 patients had Stage I disease, 128 Stage II, 237 Stage III, and nine Stage IV. [2] The studied surgery-related variables were as follows: The operation was successfully completed in all patients, 352 via a transabdominal approach, 25 via a transhiatus approach, and 80 via a transthoracoabdominal approach. The whole procedure was performed laparoscopically in 53 patients (11.6%), 189 (41.4%) underwent laparoscopic-assisted surgery, and 215 (47.0%) underwent open surgery. The median intraoperative blood loss was 200 (range, 10-1 350) mL, and the operating time 215.6±66.7 minutes. The anastomotic reinforcement time was 2 (7.3±3.9) minutes for laparoscopic-assisted surgery, 17.6±1.7 minutes for total laparoscopy, and 6.0±1.2 minutes for open surgery. [3] The studied postoperative variables were as follows: The median time to postoperative passage of flatus was 3.1±1.1 days and the postoperative gastrointestinal angiography time 6 (range, 4-13) days. The median time to postoperative oral intake was 7 (range, 2-14) days, and the postoperative hospitalization time 15.8±6.7 days. [4] The safety-related variables were as follows: In total, there were 184 (40.3%) postoperative complications. These comprised esophagojejunal anastomosis complications in 10 patients (2.2%), four (0.9%) being anastomotic leakage (including two cases of subclinical leakage and two of clinical leakage; all resolved with conservative treatment); and six patients (1.3%) with anastomotic stenosis (two who underwent endoscopic balloon dilation 21 and 46 days after surgery, the others improved after a change in diet). There was no anastomotic bleeding. Non-anastomotic complications occurred in 174 patients (38.1%). All patients attended for follow-up at least once, the median follow-up time being 10 (3-18) months. Visick grades were as follows: Class I, 89.1% (407/457); Class II, 7.9% (36/457); Class III, 2.6% (12/457); and Class IV 0.4% (2/457). Conclusion: Double and a half layered esophagojejunal anastomosis in radical gastrectomy is safe and feasible.

[Mitochondrial pyruvate carrier deficiency: 3 cases report and literature review].

Objective: To analyze the clinical and genetic features of patients with mitochondrial pyruvate carrier deficiency (MPYCD). Methods: This was a case series research. The clinical data, genetic characteristics, and glutamine treatment efficacy of 3 patients diagnosed with MPYCD at the Department of Neurology, Beijing Children's Hospital, Capital Medical University and Department of Pediatrics, Guizhou Provincial People's Hospital, from August 2019 to June 2023 were retrospectively collected. A literature search with "MPC1 gene" "MPC2 gene and" "mitochondrial pyruvate carrier deficiency" as keywords was conducted at the Wanfang Data Knowledge Service Platform, China National Knowledge Infrastructure (CNKI) and PubMed (up to June 2023). Clinical and genetic characteristics of patients with MPYCD were summarized. Results: Case 1 was a 3 years and 11 months old boy, while case 2 was a 4 years and 10 months old boy and case 3 was an 8 years and 9 months old girl. Case 2 and case 3 were siblings from one consanguineous family. All 3 patients presented with general developmental delay, growth failure and elevated serum lactate. Cranial magnetic resonance imaging (MRI) showed subtle bilateral symmetrical T2 signal hyperintensity in basal ganglia and thalamus in case 1, but normal in case 2 and 3. Trio-WES revealed case 1 harboring compound heterozygous missense variants c.208G>A (p.Ala70Thr) and c.290G>A (p.Arg97Gln) in MPC1 gene, while case 2 and 3 revealed a homozygous variant c.290G>A (p.Arg97Gln) in the same gene. All 3 cases were diagnosecl as MPYCD. Clinical symptoms including motor ability, cognition and activity endurance were improved in these 3 patients after taking glutamine for 2 years. A total of 5 articles published in English were reviewed, and no Chinese literature was found. Including these 3 cases, 15 cases were enrolled for analysis. Eleven patients carried MPC1 gene variants and 4 cases carried MPC2 gene variants. Except for 3 cases died during prenatal period, 9 of 12 enrolled born cases were onset before 6 months old. The most common clinical symptoms were mental and motor general developmental delay, microcephaly, growth failure and hypotonia. All patients had elevated blood lactate and pyruvate, but the ratio of lactate/pyruvate was normal. Seven patients performed cranial MRI, 3 exhibited non-specific changes, 2 showed bilateral symmetrical T2 signal hyperintensity in basal ganglia and thalamus, and 3 were normal. A total of 5 MPC1 gene missense variants and 2 MPC2 gene variants were identified in 15 cases. Conclusions: Onset age of patients with MPYCD is usually within 6 months. The main clinical characteristics are developmental delay, microcephaly and growth failure, accompanied by increased serum lactate and pyruvate. Glutamine supplement could lead to clinical improvements.

Correlations between the femoral neck osteotomy angle and radiologic and clinical outcomes analyzed in patients undergoing total hip replacement with metaphyseal fixation.

OBJECTIVE: This study will explore whether the femoral neck osteotomy angle (FNOA) has an effect on hip anatomical functional reconstruction and clinical outcomes after total hip arthroplasty (THA). PATIENTS AND METHODS: The study included 254 patients (296 hips) who underwent primary total hip arthroplasty using the same uncemented short stem (Tri-Lock BPS) between December 2018 and December 2019. Correlations between FNOA and the radiologic and clinical outcomes of patients were analyzed. RESULTS: Patients were divided into 3 groups according to different FNOA. FNOA ≤50° is Group A, 50°< FNOA <55° Group B, and FNOA ≥55° Group C. There were significant differences among the three groups in distal D1 (p=0.029), sitting proud (SP) (p<0.001), varus and valgus alignment (p<0.001), FO (p=0.001), and caput-collum-diaphysis angle (CCD) (p<0.001). There were significant differences in the incidence of complications among the three groups (p<0.007). There was a significant linear correlation with D1 (B=0.005, CI=0.002 to 0.008, p=0.004), SP (B=-0.266, CI=-0.286 to 0.166, p<0.001), the femoral stem varus-valgus alignment angle (B=-0.359, CI=-0.422 to -0.297, p<0.001), femoral offset (FO) (B=-0.500, CI=-0.795 to -0.205, p=0.001), and CCD (B=0.696, CI=0.542 to 0.849, p<0.001). In logistic regression analysis, inappropriate FNOA increased the risk of dislocation (OR=0.892, CI=0.812 to 0.979, p=0.016) and thigh pain (OR=0.920, CI=0.851 to 0.995, p=0.037). CONCLUSIONS: The study demonstrates the relationship between FNOA and short-term radiological and clinical outcomes of patients after THA using a Tri-Lock femoral prosthesis. Inappropriate FNOA was significantly associated with failure of hip anatomical reconstruction and a higher risk of complications.

[Clinical characteristics and short-term prognosis of 22 cases with SARS-CoV-2 infection associated acute encephalopathy].

Objective: To investigate the clinical features and short-term prognosis of patients with SARS-CoV-2 infection associated acute encephalopathy (AE). Methods: Retrospective cohort study. The clinical data, radiological features and short-term follow-up of 22 cases diagnosed with SARS-CoV-2 infection associated AE in the Department of Neurology, Beijing Children's Hospital from December 2022 to January 2023 were retrospectively analyzed. The patients were divided into cytokine storm group, excitotoxic brain damage group and unclassified encephalopathy group according to the the clinicopathological features and the imaging features. The clinical characteristics of each group were analyzed descriptively. Patients were divided into good prognosis group (≤2 scores) and poor prognosis group (>2 scores) based on the modified Rankin scale (mRS) score of the last follow-up. Fisher exact test or Mann-Whitney U test was used to compare the two groups. Results: A total of 22 cases (12 females, 10 males) were included. The age of onset was 3.3 (1.7, 8.6) years. There were 11 cases (50%) with abnormal medical history, and 4 cases with abnormal family history. All the enrolled patients had fever as the initial clinical symptom, and 21 cases (95%) developed neurological symptoms within 24 hours after fever. The onset of neurological symptoms included convulsions (17 cases) and disturbance of consciousness (5 cases). There were 22 cases of encephalopathy, 20 cases of convulsions, 14 cases of speech disorders, 8 cases of involuntary movements and 3 cases of ataxia during the course of the disease. Clinical classification included 3 cases in the cytokine storm group, all with acute necrotizing encephalopathy (ANE); 9 cases in the excitotoxicity group, 8 cases with acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) and 1 case with hemiconvulsion-hemiplegia syndrome; and 10 cases of unclassified encephalopathy. Laboratory studies revealed elevated glutathione transaminase in 9 cases, elevated glutamic alanine transaminase in 4 cases, elevated blood glucose in 3 cases, and elevated D-dimer in 3 cases. Serum ferritin was elevated in 3 of 5 cases, serum and cerebrospinal fluid (CSF) neurofilament light chain protein was elevated in 5 of 9 cases, serum cytokines were elevated in 7 of 18 cases, and CSF cytokines were elevated in 7 of 8 cases. Cranial imaging abnormalities were noted in 18 cases, including bilateral symmetric lesions in 3 ANE cases and "bright tree appearance" in 8 AESD cases. All 22 cases received symptomatic treatment and immunotherapy (intravenous immunoglobulin or glucocorticosteroids), and 1 ANE patient received tocilizumab. The follow-up time was 50 (43, 53) d, and 10 patients had a good prognosis and 12 patients had a poor prognosis. No statistically significant differences were found between the two groups in terms of epidemiology, clinical manifestations, biochemical indices, and duration of illness to initiate immunotherapy (all P>0.05). Conclusions: SARS-CoV-2 infection is also a major cause of AE. AESD and ANE are the common AE syndromes. Therefore, it is crucial to identify AE patients with fever, convulsions, and impaired consciousness, and apply aggressive therapy as early as possible.

First Report of Root-Knot Nematode Meloidogyne enterolobii on Antirrhinum majus in China.

Antirrhinum majus L. is a medicinal and ornamental herb commonly grown in China. In October 2022, A. majus plants were observed stunted in growth with yellowish leaves and containing a large number of galls on roots in a field in Nanning, Guangxi, China (N22°47'23.35″, E108°23'4.26). Ten samples were collected randomly from rhizosphere soil and roots of A. majus. Second-stage juveniles (J2) were isolated from fresh soil with a Baermann funnel, and a mean of 36 ± 2.9 per 500 cm3 of soil was recorded. Gall roots were dissected using a microscope, where 2+ 0.42 males per sample were recovered. The species was determined to be Meloidogyne enterolobii based on morphological characteristics, including the female perineal pattern, and DNA studies. Female perineal patterns and morphometric data were similar to the original description of M. enterolobii Yang and Eisenback 1983 from Enterolobium contortisilquum (Vell.) Morong in China (Yang and Eisenback 1983). The measurements of males (n = 10) included body length, 1600.7 ± 55.32 (1421.3 to 1924.3) µm; body diameter = 41.3 ± 0.80 (37.8 to 45.4) µm, stylt length = 20.5 ± 0.40 (19.1 to 22.2) µm, spicules length = 30.0 ± 0.47 (28.2 to 32.0) µm and DGO = 4.5 ± 0.3 (3.8 to 5.2) µm. Measurements of J2 (n = 20) included body length, 441.9 ± 5.42 (403.2 to 493.3) µm; body diameter = 16.6 ± 0.30 (14.4 to 8.7) µm, a = 26.8 ± 0.54 (21.9 to 31.2), c = 8.7 ± 0.27 (6.4 to 10.8), stylet length = 12.6 ± 0.17 (11.2 to 14.3) µm, DGO = 3.8 ± 0.10 (2.9 to 4.8) µm, tail length = 51.6 ± 1.27 (42.3 to 63.1) µm and hyaline tail terminus length = 11.7 ± 0.15 (10.2 to 13.1) µm. These morphological characteristics are similar to the original description of M. enterolobii (Yang and Eisenback 1983). Pathogenicity tests were conducted on A. majus 'Taxiti' plants directly germinated from seeds in a 10.5-cm-diameter pot filled with 600 ml of sterilized peat moss/sand (1:1, v/v) soil in the glasshouse. After 1 week, fifteen plants were inoculated with 500 J2/pot (nematode culture collected from the original field) and five uninoculated plants served as a control. After 45 days, aboveground parts of all inoculated plants showed symptoms similar to those observed in the field. No symptoms were observed on control plants. The RF value of the inoculated plants was determined by the method of Belair and Benoit (1996) 60 days after inoculation, and the average was 14.65. J2 were used in this test and sequenced on 28S rRNA-D2/D3, ITS, COII -16SrRNA 3 region and confirmed to be M. enterolobii. Species identification was confirmed by using polymerase chain reaction primers D2A/D3B (De Ley et al. 1999), F194/5368r (Ferris et al. 1993), C2F3/1108 (Powers and Harris, 1993). The sequences obtained GenBank accession numbers OP897743 (COII), OP876758 (rRNA) and OP876759 (ITS) were 100% similar to other M. enterolobii populations from China (MN269947), (MN648519) and (MT406251). M. enterolobii is a highly pathogenic species and has been reported in vegetables, ornamental plants, guava (Psidium guajava L.), and weeds in China, Africa and America (Brito et al. 2004; Xu et al. 2004; Yang and Eisenback 1983). The medicinal plant Gardenia jasminoides J. Ellis was also infected by M. enterolobii in China (Lu et al. 2019). Of concern is its ability to develop on crop genotypes carrying RKN resistance genes in tobacco (Nicotiana tabacum L.), tomato (Solanum lycopersicum L.), soybean (Glycine max (L.) Merr.), potato (Solanum tuberosum L.), cowpea (Vigna unguiculata (L.) Walp.), sweetpotato (Ipomoea batatas (L.) Lam.), and cotton (Gossypium hirsutum L.). Consequently, this species was added to the European and Mediterranean Plant Protection Organization A2 Alert List in 2010. This is the first natural infection report of M. enterolobii in Guangxi, China on the medicinal and ornamental herb A. majus. Acknowledgments This research was funded by the National Natural Science Foundation of China (31860492), Natural Science Foundation of Guangxi (2020GXNSFAA297076), and Guangxi Academy of Agricultural Sciences Fund, China (2021YT062, 2021JM14, 2021ZX24). References: Azevedo de Oliveira, S., et al. 2018. PLoS One 13:e0192397. Belair, G., and Benoit, D. L. 1996. J. Nematol. 28:643. Brito, J. A., et al. 2004. J. Nematol. 36:324. De Ley, P., et al. 1999. Nematol. 1:591-612. Ferris, V. R., et al. 1993. Fundam. Appl. Nematol. 16:177-184. Lu, X. H., et al. 2019. Plant Dis. 103:1434. Powers, T. O. and Harris, T. S. 1993. J. Nematol. 25:1-6 Vrain, T. C., et al. 1992. Fundam. Appl. Nematol. 15:563. Yang, B. and Eisenback, J. D. 1983. J. Nematol. 15:381.

Levels of peripheral IL-6 and CD4+ and CD8+ T cells and their prognostic significance in COVID-19.

OBJECTIVE: The aim of this study was to discuss the prognostic significance of peripheral interleukin-6 (IL-6) and CD4+ and CD8+ T cells in COVID-19. PATIENTS AND METHODS: Eighty-four COVID-19 patients were retrospectively analyzed and classified into three groups, including the moderate group (15 cases), the serious group (45 cases), and the critical group (24 cases). The levels of peripheral IL-6, CD4+, and CD8+ T cells and CD4+/CD8+ were determined for each group. It was assessed whether these indicators were correlated to the prognosis and death risks of COVID-19 patients. RESULTS: The three groups of COVID-19 patients differed significantly in the levels of peripheral IL-6 and CD4+ and CD8+ cells. The IL-6 levels in the critical, moderate, and serious groups were increased successively, but the changed levels of CD4+ and CD8+ T cells were just opposite to that of IL-6 (p<0.05). The peripheral IL-6 level increased dramatically in the death group, while the levels of CD4+ and CD8+ T cells decreased significantly (p<0.05). The peripheral IL-6 level was significantly correlated with the level of CD8+ T cells and CD4+/CD8+ ratio in the critical group (p<0.05). The logistic regression analysis indicated a dramatic increase in the peripheral IL-6 level in the death group (p=0.025). CONCLUSIONS: The aggressiveness and survival of COVID-19 were highly correlated with the increases in IL-6 and CD4+/CD8+ T cells. The fatalities of COVID-19 individuals remained at increased incidence due to elevated peripheral IL-6 levels.

First Report of Root-knot Nematode, Meloidogyne Graminicola on Brassica Juncea in China.

In southern China, the staple food rice (Oryza sativa) field is commonly rotated with brown mustard Brassica juncea. Root-knot nematodes (RKNs) are a major threat to rice production. From 2019 to 2021, B. juncea in 56 fields from 26 counties in Guangxi Province were observed with symptoms of leaf yellowing, stunting, and several hook-shaped galls on the roots. Females and egg masses of Meloidogyne sp. were found within the galls. The females, males, and second-stage juveniles (J2s) were collected, and identified with morphological and molecular characteristics and female perineal patterns. Brassica juncea was transplanted in pots and a pathogenicity test was conducted to confirm the species as Meloidogyne graminicola. In China, this is the first record of a natural infection of mustard with M. graminicola, and this finding has great importance for Chinese mustard production, since this nematode may damage mustard plants and become an additional problem for this crop.

[Outcomes at discharge of preterm infants born <34 weeks' gestation].

Objective: To investigate the incidence and trend of short-term outcomes among preterm infants born <34 weeks' gestation. Methods: A secondary analysis of data from the standardized database established by a multicenter cluster-randomized controlled study "reduction of infection in neonatal intensive care units (NICU) using the evidence-based practice for improving quality (REIN-EPIQ) study". This study was conducted in 25 tertiary NICU. A total of 27 192 infants with gestational age <34 weeks at birth and admitted to NICU within the first 7 days of life from May 2015 to April 2018 were enrolled. Infants with severe congenital malformation were excluded. Descriptive analyses were used to describe the mortality and major morbidities of preterm infants by gestational age groups and different admission year groups. Cochran-Armitage test and Jonckheere-Terpstra test were used to analyze the trend of incidences of mortality and morbidities in 3 study-years. Multiple Logistic regression model was constructed to analyze the differences of outcomes in 3 study-years adjusting for confounders. Results: A total of 27 192 preterm infants were enrolled with gestational age of (31.3±2.0) weeks at birth and weight of (1 617±415) g at birth. Overall, 9.5% (2 594/27 192) of infants were discharged against medical advice, and the overall mortality rate was 10.7% (2 907/27 192). Mortality for infants who received complete care was 4.7% (1 147/24 598), and mortality or any major morbidity was 26.2% (6 452/24 598). The incidences of moderate to severe bronchopulmonary dysplasia, sepsis, severe intraventricular hemorrhage or periventricular leukomalacia, proven necrotizing enterocolitis, and severe retinopathy of prematurity were 16.0% (4 342/27 192), 11.9% (3 225/27 192), 6.8% (1 641/24 206), 3.6% (939/25 762) and 1.5% (214/13 868), respectively. There was a decreasing of the overall mortality (P<0.001) during the 3 years. Also, the incidences for sepsis and severe retinopathy of prematurity both decreased (both P<0.001). However, there were no significant differences in the major morbidity in preterm infants who received complete care during the 3-year study period (P=0.230). After adjusting for confounders, infants admitted during the third study year showed significantly lower risk of overall mortality (adjust OR=0.62, 95%CI 0.55-0.69, P<0.001), mortality or major morbidity, moderate to severe bronchopulmonary dysplasia, sepsis and severe retinopathy of prematurity, compared to those admitted in the first study year (all P<0.05). Conclusions: From 2015 to 2018, the mortality and major morbidities among preterm infants in Chinese NICU decreased, but there is still space for further efforts. Further targeted quality improvement is needed to improve the overall outcome of preterm infants.

A 1:1 matched case-control study on dietary protein intakes and hip fracture risk in Chinese elderly men and women.

The role of protein intake in bone has been controversial. Our case-control study among Chinese elderly concluded that a higher consumption of protein, even substituted for fat, is associated with lowered hip fracture risk. Differences in protein sources, amino acids composition, gender, and calcium sufficiency may explain the inconsistency. PURPOSE: The aim of the study was to investigate the association of dietary protein intakes with hip fracture risk among Chinese elderly. METHODS: This was a 1:1 age and sex matched cross-sectional study of case-control design among 1070 pairs of elderly Chinese people aged 55 to 80 years. Patients who were newly diagnosed (within 2-week) hip fracture by X-ray were recruited from four hospitals in Guangdong Province of China. Dietary intakes were evaluated by a validated food frequency questionnaire for total protein, protein from different sources, amino acids profiles, and estimated renal acid load in diet. RESULTS: Daily average intakes of total protein were 58.1±27.0 (women) and 65.7±31.8 (men) g/d for cases, and 66.8±21.5 (women) and 72.1±24.4 (men) for controls (p<0.001). Multivariable regression indicated that, compared with the lowest quartile, the highest quartile of consumption of energy adjusted total protein [OR: 0.360 (0.206~0.630) for women and 0.381 (0.153~0.949) for men] and animal protein [0.326 (0.183, 0.560) for women and 0.335 (0.136~0.828) for men] was significantly associated with the lowered risk of hip fracture in a dose-response manner (all p for trend <0.05). A significant hip fracture risk reduction was observed in women with higher intakes of sulfur amino acids [OR: 0.464 (0.286~0.753)] and aromatic amino acids [0.537 (0.326~0.884)] but not in men. Subgroup analysis suggested that these associations were more evident in elderly with lower body mass index and dietary calcium intake less than 400 mg/d. CONCLUSIONS: A higher level of protein intake, even substituted for fat, is associated with lowered hip fracture risk.

[The prognostic value of myoglobin difference in sepsis related chronic critical illness].

Objective: To investigate the predictive value of myoglobin (Mb) for the prognosis of sepsis related chronic critical illness (CCI). Methods: Retrospective study was conducted on septic patients with the length of ICU stay equal or greater than 14 days, and sepsis-related organ failure assessment (SOFA) score equal or greater than 2 on the 14th day in ICU in the First Department of Critical Care Medicine at the First Affiliated Hospital of Sun Yat-sen University from January 2017 to March 2020. Patients' clinical and laboratory data were collected on the 1st and 14th day in ICU. The survival on day 28 in ICU was recorded. According to the myoglobin levels on day 1 and day 14, all subjects were divided into myoglobin elevation group and decline group. Kaplan-Meier survival curve was used to compare the cumulative survival rate at day 28. Cox regression analysis was used to analyze the independent risk factors of mortality. Receiver operating characteristic (ROC) curve was used to analyze the prognostic value of myoglobin. Results: A total of 131 patients with sepsis related CCI were recruited, including 58 patients in the elevation group and 73 in the decline group. The Mb level in elevation group on day 1 was significantly lower than that in decline group [172.40(59.99, 430.53) µg/L vs. 413.60(184.40, 1 328.50) µg/L, Z=3.749, P=0.000], and the Mb level on day 14 was the opposite change in two groups [483.65(230.38, 1 471.75)µg/L in elevation group vs. 132.20(76.86, 274.35)µg/L in decline group, Z=5.595, P=0.000]. Kaplan-Meier survival curve analysis showed that the 28-day cumulative survival rate of the elevation group was significantly lower than that of decline group (χ²=7.051, P=0.008). Cox ratio regression analysis suggested that elevated myoglobin was an independent risk factor for 28-day mortality in septic patients with CCI (OR=2.534, 95%CI 1.212-5.295, P=0.013). ROC curve analysis suggested that the sensitivity of myoglobin elevation in predicting mortality related to CCI within 28 days was 64.5%, and the specificity was 32.0% with area under the curve(AUC) 0.661(95%CI 0.550-0.773,P=0.007) and Jorden Index was 0.325. Conclusion: Elevated myoglobin, an independent risk factor for mortality within 28 days in ICU, can predict the prognosis of sepsis related chronic critical illness.

[Molybdenum cofactor deficiency type B manifested as Leigh-like syndrome: a case report and literature review].

Objective: To explore the phenotypes and genotypes of molybdenum cofactor deficiency type B (MoCD-B) manifested as Leigh-like syndrome. Methods: The clinical data, laboratory tests, neuroimaging and gene results of one patient diagnosed as MoCD-B at Beijing Children's Hospital and Hebei Children's Hospital in December 2018 were collected. Related literature was searched and reviewed at Wanfang Data Knowledge Service Platform, China National Knowledge Infrastructure and PubMed (up to September 2020) by using terms "MOCS2" "molybdenum cofactor deficiency" "Leigh-like syndrome,MOCS2" "molybdenum cofactor deficiency, Leigh-like syndrome". The phenotypes and genotypes of MoCD-B were summarized. Results: A 7 months and 14 days old boy with the chief complaint of "cough for 6 days, abnormal posture for 4 days and fever for 2 days" was admitted to Hebei Children' Hospital on December 2018. His abnormal posture presented as opisthotonos accompanied with dysphagia, without seizures. His previous psychomotor development was described as normal. He was born at term after an uneventful pregnancy to non-consanguineous parents. Blood test showed a slightly increased lactic acid and a significantly decreased uric acid. Urine metabolism test showed an obviously increased xanthine and hypoxanthine. Brain magnetic resonance imaging showed hyperintense signal on T2 weighted image and fluid attenuated inversion recovery in bilateral globus pallidus and pedunculus cerebri. The patient was diagnosed with Leigh-like syndrome. No obvious improvement was achieved after cocktail therapy and symptomatic treatment. The whole exome sequencing showed that the patient carried a homozygous variant of MOCS2 gene, c.19G>T(p.Val7Phe), which was a previously reported pathogenic site in the literature and could cause MoCD-B. His parents carried a heterozygous variant respectively. A total of 41 MoCD-B cases with MOCS2 gene variants were collected through literature review and our study, among which 30 cases had full medical records. The onset ages of 23 (77%) cases were in neonate, manifesting with severe encephalopathy, including neonatal-onset intractable seizures, developmental delay, laboratory abnormalities included very low levels of serum and urinary uric acid, increased urinary levels of xanthine and hypoxanthine. Cranial imaging showed cerebral atrophy, cystic encephalomalacia, etc. The onset ages of 7 patients varied from 5 months to 23 years. Four cases had normal psychomotor development before disease onset. Neurological disorders appeared acutely or exacerbated after external triggers and all of them had basal ganglia involvement. Among the 30 cases, 3 cases had a relatively milder phenotype with the ability of brief communication and walking without or with support. Conclusions: Molybdenum cofactor deficiency is a rare disease. Most cases had severe phenotypes and poor outcomes, but some cases may have mild phenotype. MoCD-B caused by MOCS2 gene variants may manifest as Leigh-like syndrome with a normal psychomotor development before the trigger of infection strike. Hypouricemia, xanthinuria and hypoxanthinuria can be indicators of the disease. The presence of MOCS2 gene variants would confirm a final diagnosis.

[Analysis on clinical efficacy, safety and economy of Shaobei injection and elastic band ligation in the treatment of grade II or III hemorrhoids].

Objective: Currently, various treatments such as hemorrhoidectomy, ligation and sclerotherapy injection can be applied in grade II or III hemorrhoids. This study aims to compare the clinical efficacy, safety and economy between Shaobei injection and elastic band ligation in treating patients with grade II or grade III hemorrhoids. Methods: A retrospective cohort study was used. Clinical data of 60 patients with grade II or grade III hemorrhoids at Department of Anorectal Surgery of the Sixth Affiliated Hospital, Sun Yat-sen University between January 2019 and October 2019 were collected. Patients were divided into two groups according to surgical methods. Patients in the Shaobei group received Shaobei injection (n=28), and those in the ligation group received elastic band ligation (n=32). Inclusion criteria: (1) diagnosis of grade II or III hemorrhoid; (2) application of Shaobei injection or elastic band ligation; (3) age between 18-75 years old. Exclusion criteria: (1) comorbidity with anal fissure, anal fistula, anal sinusitis or other perianal diseases; (2) patients with mental disorder or poor compliance; (3) incomplete clinical or follow-up data. Recurrent rate, postoperative pain, anal edema, anal distension, total cost of hospitalization, length of hospitalization, and postoperative life quality EQ-5D-3L score were compared between the two groups at postoperative 6-month. Results: No significant difference was observed in the baseline data (including Nystrom hemorrhoid symptom score) between the two groups (all P>0.05), except gender ratio [male proportion: Shaobei 75% (21/28) vs. ligation 37.5%(12/32), χ(2)=8.485, P=0.004]. No significant difference in recurrent rate was found between the two groups [14.3% (4/28) vs. 9.4% (3/32), χ(2)=0.035, P=0.851]. Compared to the ligation group, Shaobei group showed less pain at postoperative day 1 [VAS median (range): 2 (1-6) vs. 3 (1-7), Z=2.814, P=0.005] and postoperative day 7 [VAS median (range): 0 (0-2) vs. 1 (0-4), Z=3.149, P=0.002]; lower anal edema ratio at postoperative day 1 [10.7% (3/28) vs. 34.4% (11/32), Z=4.673, P=0.037]; lower anal distension ratio at postoperative day 1 [7.1% (2/28) vs. 28.1% (9/32), Z=4.391, P=0.048]; less hospitalization cost [(6343.5±1444.1) yuan vs. (10 587.1± 1719.0) yuan, t=12.515, P<0.001] and shorter postoperative hospital stay [median (range): 1 (1-5) days vs. 3 (1-6) days, Z=5.879, P<0.001]. The EQ-5D-3L scores of two groups were significantly improved six months after treatment [Shaobei group: (0.90±0.16) vs. (0.73±0.14); ligation group: (0.91±0.13) vs. (0.74±0.10); both P<0.001], while there was no statistically significant difference between the two groups (t=0.130, P=0.897). No complications such as massive hemorrhage, infection, iatrogenic anal fistula, rectal stricture and local induration occurred after the injection. Conclusions: Shaobei injection is effective and safe in treating grade II or III hemorrhoids. Compared with elastic band ligation, it can reduce morbidity of complications and hospitalization expenses.

LncRNA GAS5 induces chondrocyte apoptosis by down-regulating miR-137.

OBJECTIVE: Long non-coding RNA (lncRNA) participates in the pathogenesis of human knee osteoarthritis (KOA). Growth arrest specificity 5 (GAS5) is a member lncRNA, but its role in pathological regulation of KOA is still unknown. This study aims to explore the mechanism of GAS5 in KOA on chondrocyte apoptosis and other pathological processes. PATIENTS AND METHODS: The serum and cartilage tissues were collected from 35 patients with KOA and 30 patients with traumatic amputation admitted to our hospital from April 2016 to April 2020. The expressions of GAS5 and miR-137 were detected and analyzed. Chondrocytes were extracted from cartilage tissues of KOA patients, and the genes were regulated by transfection. Then, the cells were detected, including apoptosis, apoptosis-related proteins (caspase-3, Bax/Bcl-2), and proliferation. The targeting relationship between GAS5 and miR-137 was verified. RESULTS: GAS5 was up-regulated in serum and cartilage tissues of KOA patients, and down-regulation of GAS5 could inhibit the apoptosis process of chondrocytes and promote proliferation. MiR-137 was down-regulated in samples of KOA patients and was negatively regulated by GAS5. GAS5 induced apoptosis of chondrocytes and inhibited its proliferation through targeted down-regulating miR-137. CONCLUSIONS: GAS5 is up-regulated in KOA serum, cartilage tissues and cells, and can induce chondrocyte apoptosis through down-regulating miR-137.

[HIV positive rate from different detection methods in medical institutions in China: a Meta analysis].

Objective: To analyze the HIV positive detection rate from different detection channels in Chinese medical institutions. Methods: A Meta-analysis was conducted. First of all, the literature on HIV testing of medical institutions in China was systematically searched on China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, VIP Information Chinese journal Service platform and PubMed. Secondly, a self-made information table was used to collect the basic information, HIV positive number and test number of the literature. Finally, R 4.0.2 software was used to calculate the pooled HIV detection rate and 95%CI of the whole population, detection approaches subgroups and regions subgroups, and then the forest map was drawn. Funnel plot was used to analyze publication bias. Results: A total of 45 studies which covered 22 provinces. Meta analysis showed that the pooled HIV positive rate was 0.82‰ (95%CI: 0.62‰-1.04‰). Subgroup analysis showed that the HIV positive rate of STD outpatient was the highest (3.01‰ (95%CI: 1.76‰-4.58‰), followed by other patients (1.43‰ (95%CI: 1.00‰-1.93‰)). The HIV positive rate of western China was the highest (1.14‰ (95%CI: 0.72‰-1.63‰)). The HIV positive rate in 2008-2017 was higher than in 2000-2007. The Egger test indicated no publication bias (t=-0.737, P=0.465). Conclusion: The HIV positive detection rate of patients in medical institutions in China was at a low level, but the positive rate of patients in STD clinics was relatively high. Therefore, the HIV testing should be further expanded in this population. Secondly, HIV screening should be strengthened for other patients.

[Clinical characteristics of patients with indication of cardiac implantable electronic devices implantation complicating with acute pulmonary thromboembolism].

Objective: To investigate the clinical characteristics of inpatients with the indication of cardiac implantable electronic devices (CIED) therapy and combined acute pulmonary thromboembolism (APTE). Methods: We retrospectively screened 8 641 inpatients who admitted with the indication of CIED implantation in Fuwai Hospital from January 2014 to May 2019. The clinical characteristics, management strategies and clinical outcome were analyzed for patients diagnosed as APTE. Results: APTE were identified in 45 (5‰) patients in this cohort, there were 18(40%) male patients, the average age was (73±8) years old and body mass index was (27±10) kg/m2.Thirty-two (70%) patients were at intermediate-risk and 13 (30%) at low-risk. Anti-coagulation therapy was initiated in 38(84%) patients, and 30 patients underwent CIED implantation (27 pacemaker, 2 CRT and 1 ICD). No postoperative bleeding or pocket hematoma were detected in the 23 patients taking anticoagulation medication before implantation. During an average of (30±7) months' follow up, thrombus was dissolved in 20 patients, hemorrhage complications were observed in 2 patients (1 cerebral hemorrhage and 1 hematuria), anticoagulation therapy was discontinued in these 2 patients. Among 15 patients without immediate CIED implantation and treated with anticoagulation therapy during hospitalization, 2 patients developed complete paroxysmal Ⅲ° atrioventricular block, and recovered after therapy during hospitalization. Seven patients were re-hospitalized for CIED implantation due to bradycardia. Five patients died during follow-up (3 sudden cardiac death, 1 APTE combined with cerebral infarction, and 1 pulmonary infection). Conclusion: APTE is not rare in patients with the indication of CIED implantation, CIED implantation and anti-coagulation therapy are safe for these patients, and transient atrioventricular block could be detected in APTE patients.

MiR-1301 promotes adipogenic and osteogenic differentiation of BMSCs by targeting Satb2.

OBJECTIVE: Bone marrow mesenchymal stem cells (BMSCs) have the ability to differentiate into several cell lines and are critical for skeletal microenvironment and bone development. MiR-1301 is involved in multiple pathological and physiological processes. However, miR-1301's role in BMSCs adipogenic and osteogenic differentiation remains unclear. MATERIALS AND METHODS: Rat BMSCs were isolated and randomly divided into control group, miR-1301 group, and miR-1301 siRNA group followed by analysis of the expression of miR-1301, Bax, Bcl-2, UNX2, and OPN, as well as FABP4 and PPARγ2 by Real Time-PCR. Cell proliferation was assessed by MTT assay and the relationship between miR-1301 and Satb2 was evaluated by the Dual-Luciferase reporter assay. Satb2 expression was detected by Western blot. RESULTS: The pcDNA-miR-1301 plasmid was transfected into BMSCs to upregulate the expression of miR-1301, which promoted cell proliferation, decreased Bax expression, and increased Bcl-2 expression and ALP activity. In addition, it also elevated the expression of RUNX2 and OPN and decreased the expression of FABP4, PPARγ2, and Satb2. Compared with the control group, the difference was statistically significant (p<0.05); Satb2 was the target gene of miR-1301. MiR-1301 siRNA transfected into BMSCs down-regulated miR-1301 expression, inhibited cell proliferation, increased Bax expression and decreased Bcl-2 expression and ALP activity. Meanwhile, miR-1301 siRNA also reduced RUNX2 and OPN expression and increased expression of FABP4, PPARγ2 and Satb2. The difference was statistically significant compared with control group (p<0.05). CONCLUSIONS: Regulation of miR-1301 expression in BMSCs can improve BMSCs proliferation and regulate their adipogenic and osteogenic differentiation by regulating Satb2.

[Homologous modeling and binding ability analysis of Spike protein after point mutation of severe acute respiratory syndrome coronavirus 2 to receptor proteins and potential antiviral drugs].

OBJECTIVE: To explore the natural mutations in Spike protein (S protein) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the changes of affinity between virus and associated receptors or drug molecules before and after the mutation based on whole length sequencing results. METHODS: In the study, the bioinformatics analysis of all the published sequences of SARS-CoV-2 was conducted and thus the high frequency mutation sites were affirmed. Taking advantages of PolyPhen-2, the functional influence of each mutation in S protein was prospected. The 3D homologous modelling was performed by SWISS-MODEL to establish mutated S protein structural model, in which the protein-docking was then implemented with angiotensin-converting enzyme 2 (ACE2), dipeptidyl peptidase-4 (DPP4) and aminopeptidase N (APN) by ZDOCK, and the combining capacity of each mutated S protein evaluated by FiPD. Finally, the binding ability between mutated S proteins and anti-virus drugs were prospected and evaluated through AutoDock-Chimera 1.14. RESULTS: The mutations in specific region of S protein had greater tendency to destroy the S protein function by analysis of mutated S protein structure. Protein-receptor docking analysis between naturally mutated S protein and host receptors showed that, in the case of spontaneous mutation, the binding ability of S protein to ACE2 tended to be weakened, while the binding ability of DPP4 tended to be enhanced, and there was no significant change in the binding ability of APN. According to the computational simulation results of affinity binding between small molecular drugs and S protein, the affinity of aplaviroc with S protein was significantly higher than that of other small molecule drug candidates. CONCLUSION: The region from 400-1 100 amino acid in S protein of SARS-CoV-2 is the mutation sensitive part during natural state, which was more potential to mutate than other part in S protein during natural state. The mutated SARS-CoV-2 might tend to target human cells with DPP4 as a new receptor rather than keep ACE2 as its unique receptor for human infection. At the same time, aplaviroc, which was used for the treatment of human immunodeficiency virus (HIV) infection, may become a new promising treatment for SARS-CoV-2 and could be a potential choice for the development of SARS-CoV-2 drugs.