Objective: To optimize the ultrahigh pressure extraction ( UPE) process for forsythoside from Forsythia suspensa by Box-Behnken experimental design. Methods:On the basis of single factor screening, a three-factor and three-level Box-Behnken ex-perimental design was employed with liquid/material ratio( X1 ) , extraction pressure ( X2 ) and extraction time ( X3 ) as the independent variables. The response variable was the extraction yield of forsythoside. Results:The optimal UPE conditions for forsythoside were as follows:the ratio of solvent to material (ml·g-1) was 70, the extraction pressure was 151 MPa, and the extraction time was 114 s. With the optimal extraction process, the extraction yield of forsythoside was 13. 15 mg·g-1 . Conclusion:As a novel extraction tech-nology for Chinese herbal medicines, UPE procedure has higher extraction yield, lower extraction temperature, shorter extraction time and less power consumption, which provides a brand-new method for the extraction of forsythoside from Forsythia suspensa.

SHIV-CN97001 played an important role in assessing the immune effect and strategy of the AIDS vaccine which included genes of the predominant prevalent HIV-1 strain in China. In this study, SHIV-CN97001 was in vivo passaged serially to construct pathogenic SHIV-CN97001/rhesus macaques model. To identify variation in the gp120 region of SHIV-CN97001 during passage, the fragments of gp120 gene were amplified by RT-PCR from the plasma of SHIV-CN97001 infected animals at the peak viral load time point and the gene distances (divergence, diversity) were calculated using DISTANCE. The analysis revealed that the genetic distances of SHIV-CN97001 in the third passage animals were the highest during in vivo passage. It had a relationship between viral divergence from the founder strain and viral replication ability. The nucleic acid sequence of the V3 region was highly conservative. All of the SHIV-CN97001 strains had V3 loop central motif (GPGQ) and were predicted to be using CCR5 co-receptor on the basis of the critical amino acids within V3 loop. These results show that there was no significant increase in the genetic distance during serial passage, and SHIV-CN97001 gp120 gene evolved toward ancestral states upon transmission to a new host. This could partly explain why there was no pathogenic viral strain obtained during in vivo passage.

Objective To compare the mRNA level of macrophage inflammatory protein-3α(MIP-3α) in 3 different mucosal epithelial cell lines. Methods RNA standards were prepared by in vitro transcription. One-step real-time RT-PCR was established and optimized using TaqMan EZ RT-PCR Core Reagents with TaqMan probes and primers specific to human MIP-3α mRNA sequence. The specificity of one-step real-time RT-PCR method was confirmed by sequencing the PCR products. The sensitivity and reproducibility of the method was examined by repeating the test 8 times with the same sample. Results The one-step real-time RT-PCR with a wide detection range is sensitive, reproducible. It was found that MIP-3α mRNA level in Caco-2 and T-84 cells was much higher than that in the HeLa cells. Conclusion High level of MIP-3α mRNA could be found in mucosal epithelial cells and difference in transcription level of MIP-3α may exist in epithelial cells from different mucosa.

Objective To evaluate the efficacy and safety of retrograde flexible ureteroscopic lithotripsy ( RFUL) with holmium:YAG laser for the treatment of renal stones with a large series.Methods The data of 466 patients who underwent RFUL with holmium:YAG laser for the treatment of kidney stones between January 2013 and December 2013 were collected.The maximum diameter of stone was 23 ±16 mm.The stone free rate, complications, retreatment rate were evaluated.Results Out of the 466 patients, the mean operative time and postoperative hospital stay were 33.5 ±18.8 min and 2.3 ±2.0 d.The stone free rate was 67.6% ( 315/466 ) after single procedure, which increased to 69.5% ( 324/466 ) via re-treatments after 3 months.The retreatment rate was 4.3%(20/466), with a total of 493 RFUL procedures performed and 1.06 times per patient.There were 67 (14.4%) cases undergoing complications.Five cases had false passage of ureter orifice causing slight ureteral wall injuries.Steinstrasse occurred in 9 cases, ureteral perforation in 3 cases and perirenal hematoma in 1 case.Overall postoperative fever rate was 10.7%(50/466) with urosepsis in 16 cases (3.4%).When the stone size was≤10, 11-20, 21-30, 31-40, and >40 mm, the stone free rates after a single procedure were 97.0% ( 65/67 ) , 84.2% ( 160/190 ) , 63.1%(70/111), 29.2%(14/48), 12.0%(6/50) (P<0.001), and the postoperative fever rates were 1.5%(1/67), 9.5%(18/190), 13.5% (15/111), 14.6% (7/48), 18.0% (9/50), respectively. The postoperative fever rates were 19.3%(27/140) and 7.1%(23/326) (P<0.001) in the patient with positive and negative preoperative urine leukocyte, and 22.0% ( 22/100 ) and 7.7% ( 28/366 ) ( P <0.001) in patients with positive and negative preoperative urine culture.Conclusions RFUL with holmium:YAG laser is a safe and effective treatment for kidney stones.The postoperative fever rate would increase and stone free rate would reduce with the increased stone size.

Rational drug use and patient medication safety have always been key to pharmaceutical management.The authors introduced the practice of informationization management of rational drug use in the medical alliance of Luohu Hospital Group.The main measures refer to building an informationization platform for drug purchasing and supply management, dispensing management, medication intervention and medication guidance.This practice can realize closed-loop drug management, achieving full-range supervision and traceability for drugs and higher patient drug use safety.

The WRKYs are a group of plant-specific transcription factors that play important roles in defense responses. In this study, we silenced 2 GmWRKY33B homologous genes using a bean pod mosaic virus (BPMV) vector carrying a single fragment from the conserved region of the GmWRKY33B genes. Silencing GmWRKY33B did not result in morphological changes. However, significantly reduced resistances to Pseudomonas syringae pv. glycinea (Psg) and soybean mosaic virus (SMV) were observed in the GmWRKY33B-silenced plants, indicating a positive role of the GmWRKY33B genes in disease resistance. Kinase assay showed that silencing the GmWRKY33B genes significantly reduced the activation of GmMPK6, but not GmMPK3, in response to flg22 treatment. Reverse transcriptase PCR (RT-PCR) analysis of the genes encoding prenyltransferases (PTs), which are the key enzymes in the biosynthesis of glyceollin, showed that the Psg-induced expression of these genes was significantly reduced in the GmWRKY33B-silenced plants compared with the BPMV-0 empty vector plants, which correlated with the presence of the W-boxes in the promoter regions of these genes. Taken together, our results suggest that GmWRKY33Bs are involved in soybean immunity through regulating the activation of the kinase activity of GmMPK6 as well as through regulating the expression of the key genes encoding the biosynthesis of glyceollins.
Glycine max/genetics* ; Disease Resistance/genetics* ; Biological Assay ; Dimethylallyltranstransferase ; Gene Silencing

A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.

【Objective】 To analyze ABO system hemolytic disease of the fetus and newborn (HDFN) and its influencing factors in Obstetrics Department of our hospital. 【Methods】 The blood samples of 1 040 neonates and their mothers in the obstetric department of our hospital were retrospectively analyzed from September 2022 to January 2023, including ABO and RhD blood group of the neonates and mothers, as well as 3 tests of HDFN, Hb, total bilirubin (TBIL) and indirect bilirubin(IBIL) of the neonates. Relevant clinical data of the neonates and mothers were collected, including maternal and neonatal age, neonatal sex, maternal pregnancy history, gestational age and delivery mode, and their influences on ABO-HDFN were analyzed. 【Results】 Among 1 040 HDFN samples, 298 were ABO incompatibility, among which 113 were HDFN positive, with a positive rate of 37.9% (113/298); the positive rate of HDFN in neonates born to mothers with type O was significantly higher than that in neonates born to mothers with type A and B (71.4% vs 8.2%, P<0.05); the positive rate of HDFN in neonates with antigen-A incompatibility was significantly higher than that in neonates with antigen-B incompatibility (48.7%vs 26.7%, P<0.05), which was the highest in neonates with O-A incompatibility [83.6% (61/73)], followed by O-B incompatibility [58.2% (39/67)]. There was no significant difference in Hb and bilirubin among the other groups except for the difference of Hb between the O-A incompatibility HDFN positive group and the HDFN negative group [(145.0±16.0) vs(153.4±13.2), P<0.05)]. The levels of Hb, TBIL and IBIL in the "direct antiglobulin test+ indirect antiglobulin test+release test+" group were significantly different from those in the HDFN negative group[(144.9±21.6) vs (153.3±13.2), P <0.05; (36.9±11.8) vs (29.6±6.1), P<0.05; (30.6±12.7) vs (23.0±6.9), P<0.05, respectively]. Logistic regression analysis showed that maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN. 【Conclusion】 ABO-HDFN occurred mainly in neonates born to O-type mothers, and the positive rate was the highest in neonates with O-A incompatibility. The severity of HDFN had little relationship with the mother-neonate blood type, but had relationship with the result of 3 tests of HDFN. Maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN.

Background::Understanding the characteristics of newly diagnosed primary human deficiency virus-1 (HIV-1) infection in the context of the post-antiretroviral therapy era and HIV drug prophylaxis is essential for achieving the new target of 95-95-95-95 by 2025. This study reported the characteristics of newly diagnosed primary HIV-1 infection in Shenzhen.Methods::This is a real-world retrospective study. Eighty-seven newly diagnosed primary HIV-1-infected patients were recruited from January 2021 to March 2022 at the Third People’s Hospital of Shenzhen. Demographic, epidemiological, diagnostic, drug resistance, and medical data were described and analyzed.Results::Overall, 96.6% (84/87) of the newly identified primary HIV-1-infected patients were male, including 88.5% (77/87) men have sex with men (MSM), with a median age of 29.0 years (Q 1-Q 3: 24.0-34.0 years); of these, 85.1% (74/87) reported high-risk sexual behaviors with casual partners. The rate of condom usage was only 28.7% (25/87). The overall rate of pre-exposure prophylaxis (PrEP) and post-exposure prophylaxis (PEP) was 8.0% (7/87, including 4 PrEP and 3 PEP cases) around the potential exposure, although 41.4% of the patients had prior awareness of such interventions. Moreover, only 19.5% (17/87) had previously used PrEP or PEP. Of those, 58.8% (10/17) of the patients obtained drugs from the internet, and only 35.3% (6/17) reported good compliance. A total of 54.0% (47/87) of subjects were diagnosed by the HIV nucleic acid test. Acute retroviral syndrome appeared in 54.0% (47/87) of patients. The prevalence of transmitted drug resistance (TDR) mutation was 33.9% (19/56), including 6 (10.7%) against nucleoside reverse transcriptase inhibitor (NRTI) plus non-nucleoside reverse transcriptase inhibitor (NNRTI), 8 (14.3%) against NNRTI, and 5 (8.9%) against protease inhibitor (PI) only. Conclusions::Owing to the low utilization rate and incorrect usage of PrEP and PEP, massive efforts are needed to promote HIV-preventive strategies in the MSM population. The extremely high prevalence of TDR mutation in this population implies the need for future pretreatment drug resistance surveillance.

Synthetic biology is an emerging interdisciplinary research field. By designing and constructing new or re-designing the existing natural systems, it confers them novel functions, which do not exist in nature. Owing to the predictability and controllability, synthetic biology attracts more and more interest from biologists, physicists, and engineers. Synthetic biology approaches not only can be widely used for biotechnological applications but also can be used to study complex biological systems to address fundamental questions. Here, we reviewed the recent studies following the concept of "build-to-understand", particularly, the studies to understand intracellular network structure, cell physiology, the behavior of multicellular populations, and ecosystems.